Biology subjects

Ghosh, S.

Publications and source records attributed to Ghosh, S..

At least 91 records · Page 5Linked to original sources

bioRxiv2025

Deep Coupled Kuramoto Oscillatory Neural Network (DcKONN): A Biologically Inspired Deep Neural Model for EEG Signal Analysis

Deep neural networks applied to signal processing tasks often need specialized architectural mechanisms to capture the temporal history of input signals. Traditional approaches include recurrent loops between layers, gated units, or tapped delay lines. However, biological brains exhibit much richer dynamics, characterized by activity across multiple frequency bands (alpha, beta, gamma, delta) and phenomena such as phase locking and synchronization. Standard Recurrent Neural Networks (RNNs) are limited in their ability to represent these complex dynamical features. In this work, we introduce a novel framework called the Deep Coupled Kuramoto Oscillatory Neural Network (DcKONN), which leverages networks of nonlinear Kuramoto oscillators trained in a deep learning paradigm. The DcKONN architecture has been applied to EEG signal classifier task. Simulation results demonstrate that the proposed oscillatory neural networks achieve superior or comparable classification accuracy compared to existing state-of-the-art models. Beyond performance improvements, these models also provide valuable neurobiological insights by naturally incorporating oscillatory dynamics into their architecture.

bioengineering↗

bioRxiv2025

Spatio Temporal Attentional EEGNet: An Enhanced Deep Learning Model for Cognitive Workload Detection

Deep learning has emerged as a powerful tool for extracting meaningful patterns from electroencephalography (EEG) signals, particularly for mental workload recognition in cognitive neuroscience. This study proposes STA-EEGNet (Spatio Temporal Attentional EEGNet), an enhanced variant of the EEGNet architecture that integrates spatio temporal attention mechanisms to improve EEG signal decoding. The model is trained and evaluated on the STEW: Simultaneous Task EEG Workload Dataset, demonstrating superior classification accuracy compared to existing approaches. Heatmap analyses of key convolutional filters in TSA-EEGNet network during testing reveal distinct spatiotemporal activation patterns associated with varying workload levels. High workload is characterized by increased activation in discriminative filters and reduced or inverted responses in suppressive filters, primarily over frontocentral and parietal regions. These findings highlight TSA-EEGNets ability not only to achieve state-of-the-art performance but also to provide interpretable insights into the neural dynamics underlying cognitive workload.

bioengineering↗

bioRxiv2025

Tretinoin, a Vitamin A derivative, exerts antiviral effects against Chikungunya virus infection through the nuclear retinoid receptor signaling pathways

Chikungunya virus (CHIKV) has reemerged as a global pathogen causing serious public health threat and socio-economic damage, particularly due to the absence of specific antivirals. Retinoids have been reported to exhibit antiviral potentials against multiple viral infections. In the present study, Tretinoin (TR) or all-trans retinoic acid (ATRA) was evaluated for its anti-CHIKV activity using in vitro, in vivo, and ex vivo models. TR was able to impede CHIKV infection efficiently in Vero and physiologically relevant muscle cells, C2C12, with drastic reduction in the viral RNA, proteins and progeny formation. The IC50 of TR against CHIKV was estimated to be 20.05 {micro}M and 19.71 {micro}M in Vero and C2C12 cells, respectively. In addition, the inhibition was effective during entry as well as in the early stages of post CHIKV-infection. Notably, TR showed remarkable virucidal activity. Next, global transcriptome profile demonstrated that the signaling by Retinoic acid pathway was highly upregulated after CHIKV infection and Retinoic acid receptors (RAR), specifically RAR-{beta} are the key mediators for the anti-CHIKV effect of TR. Further, the drug was also capable to modulate CHIKV-induced inflammatory responses by reducing the phosphorylated MAPKs, NF-{kappa}B and proinflammatory cytokines. Interestingly, TR protected C57BL/6 mice from CHIKV challenge by diminishing viral burden leading to reduced clinical scores and better survival. Similar observation was also noticed in the ex vivo model of CHIKV-infected hPBMC-derived monocyte-macrophage populations. In conclusion, this work demonstrated the repurposing potential of TR against CHIKV infection for the first time, encouraging its clinical validation towards future therapeutics. IMPORTANCECHIKV is an arbovirus causing Chikungunya Fever (CHIKF) leading to debilitating arthralgia and myalgia. The unavailability of licensed antivirals or potent vaccines against CHIKV encourages extensive research to find a therapeutic cure. Retinoids, the derivatives of vitamin A, have been repurposed as antiviral agents against a broad range of viruses; however, their efficacy against alphaviruses remains unexplored. Accordingly, Tretinoin (TR), also known as all-trans retinoic acid (ATRA), was validated for anti-CHIKV efficacy in this investigation. Significant perturbation of CHIKV titer through the modulation of Retinoic acid receptors (RAR) was observed upon drug-treatment. Further, TR reduced major inflammatory markers (MAPKs and proinflammatory cytokines) supporting its anti-inflammatory effect. Further, its anti-CHIKV potential was validated in the mouse model and hPBMC-derived monocyte-macrophage cells, indicating the pre-clinical efficacy. Finally, this is the first study to report the anti-CHIKV property of Tretinoin using in vitro, in vivo, and ex vivo approaches suggesting its repurposing potential.

microbiology↗

bioRxiv2025

De novo heterozygous variants in EHMT2 genocopy Kleefstra syndrome via loss of G9a methyltransferase activity

EHMT1 and EHMT2 genes encode human euchromatin histone lysine methyltransferase 1 and 2 (EHMT1 alias GLP; EHMT2 alias G9a) that form heteromeric GLP/G9a complexes with essential roles in epigenetic regulation of gene expression. While EHMT1 haploinsufficiency has been established as the cause of Kleefstra syndrome 1, the pathogenesis of G9a dysfunction in human disease remains largely unknown. We identified seven de novo EHMT2 variants in patients with clinical presentation, episignatures, histone modifications and transcriptomic profiles similar to those of Kleefstra syndrome 1. In vitro studies revealed that these variants encode for structurally stable G9a proteins that are catalytically incompetent due to aberrant interactions either with histone H3 tail or with S-adenosylmethionine. Heterozygous mice carrying a patient-derived variant exhibited growth retardation, facial/skull dysmorphia and aberrant behavior. Here we report pathogenic EHMT2 variants that likely exert dominant-negative effect on GLP/G9a complexes and thus genocopy the EHMT1 haploinsufficiency via a distinct molecular mechanism, defining an autosomal dominant EHMT2-related Kleefstra syndrome.

genetics↗

bioRxiv2025

RB1 gene mutations and their association with Endometrial Cancer

Retinoblastoma gene (RB1) mutation has been reported in lung cancer, retinoblastoma and cervical cancer etc. Here, we elucidate the involvement of RB1 mutations and their regulation in endometrial cancer. Analysis of mutation data of 547 endometrial cancer samples revealed that 12% of samples harboured RB1 mutations. However, 26% of the patients aged between 30 and 50 years harboured RB1 mutations, compared to only 10% in the older age group. Further, in silico interaction studies and structural analysis of these novel RB1 mutations and E2F revealed possible physiological relevance. We found 14,117 genes were significantly mutated, and 5,770 genes were differentially expressed in RB1-altered patients. Pathway analysis showed a significant correlation between RB1 mutations and estrogen receptor mutations. Different cancer-related pathways were also altered in RB1-mutated conditions. More importantly, a positive link between RB1 mutations and HPV infection pathways was observed, indicating HPV infections might induce RB1 mutations in endometrial cancer, which could be prevented by early vaccination. Our findings suggest RB1 mutations as a potential contributor to endometrial cancer in younger women, highlighting the importance of viral co-factors in risk assessments, advocating HPV vaccination in individuals with RB1 mutations.

cancer biology↗

bioRxiv2025

Clustering Gene Co-expression Using 2D Contour Analysis and Autoencoder-based Embedding

Identifying co-expressed genes is crucial for understanding biological processes; however, common methods such as Pearson and Spearman correlation rely on assumptions of linearity and monotonicity, respectively, that may not hold for complex biological data. To address these limitations, we propose a framework that captures visual features of gene expression profiles without relying solely on correlation-based methods. Our approach involves converting 3D gene expression data, which contain rich information on gene envelopes over time, into 2D contours that retain important visual information. We then train an autoencoder on the 2D contour images and generate embeddings from them, followed by clustering on the generated embeddings. We also introduce a new clustering algorithm, TAHC, based on hierarchical clustering, which performs better than existing methods in higher dimensions while using cosine similarity. We apply this framework to the Percellome database, which contains gene expression data with two variables--time and dosage--across various experimental conditions (e.g., tissue type). The resulting clusters exhibit good visual coherence, with an overall average Pearson correlation coefficient of 0.81, demonstrating the effectiveness of our approach.

pharmacology and toxicology↗

bioRxiv2025

A user-friendly goniometer-compatible fixed-target platform formacromolecular crystallography at synchrotrons

Fixed-target platforms provide convenient support for microcrystals during serial X-ray crystallography studies using synchrotron radiation. Here, we describe a simple, user-friendly 3D-printed support where the crystals are sandwiched between two layers of thin X-ray transparent membrane resulting in very low scattering background. The platform is compatible with magnetic mounting onto the standard goniometer of macromolecular crystallography beamlines. Our design utilizes a 96-well frame that facilitates hanging-drop experiments directly on the membrane using conventional crystallization plates, thereby eliminating multiple pipetting and crystal handling steps. Crystals can be enclosed into a sandwich and packed into cassettes, preventing the risk of the sample drying out during room-temperature transportation to synchrotron sources. The versatility of the platform is demonstrated by five structures solved using different crystallization- and data-collection strategies. Single crystal rotational-crystallography at both room- and cryogenic-temperatures using large crystals of lysozyme is shown. On-chip microcrystallization is illustrated by use of a photosynthetic reaction center as an example. Finally, serial crystallography data collection at room-temperature from microcrystals of photosynthetic reaction center as well as cytochrome c oxidase crystallized in lipidic cubic phase is presented.

biophysics↗

bioRxiv2025

Heterotypic intercellular adhesion tunes efficiency of cell-on-cell migration

Cell migration across epithelial barriers occurs in diverse developmental, immunological, and pathological contexts. Here we investigate the contribution of heterotypic adhesion between migrating cells and epithelial "substrate" cells to transepithelial migration. Using an in silico model inspired by the migration of primordial germ cells across the midgut epithelium in the Drosophila embryo, we show that heterotypic adhesion modulates migration efficiency in a non-monotonic manner, revealing the existence of an optimal adhesion regime. Consistent with this prediction, in vivo overexpression of E-cadherin in germ cells accelerated their exit from the midgut relative to controls. Beyond providing experimentally testable predictions, our model integrates and explains previous observations on the role of heterotypic adhesion in cell-on-cell migration, offering a framework for understanding transepithelial migration across biological contexts. Significance StatementCell adhesion is important for cell migration, and when cells migrate on a substrate of other cells (rather than on extracellular matrix), the adhesive properties of both cell types must be considered. However, whether and how dynamic changes in adhesion regulate cell-on-cell migration remains unclear. Here we approach this problem using transepithelial migration of Drosophila embryonic germ cells as a case study. We develop an in silico model of the migration process that predicts an optimal level of adhesion between migrating cell and substrate cell to achieve efficient migration. In vivo live imaging and genetic manipulation experiments uphold the predictions of this model. This suggests that adhesion is not a simple on/off binary parameter regulating migration.

biophysics↗

bioRxiv2025

Evolutionary innovation through fusion of sequences from across the tree of life

Novel genes arise through multiple mechanisms, including gene duplication, gene fusion and horizontal gene transfer (HGT). While HGT has increasingly been documented in animals, the post-transfer evolutionary fate of horizontally-acquired genes is less well understood. We hypothesized that fusion with endogenous sequences in animal genomes might generate what we call "HGT-chimeras": genes with regions of non-metazoan and metazoan descent in the same open reading frame. To test this hypothesis, we developed a molecular phylogenetics pipeline that enables the identification of HGT-chimeras. We applied our pipeline to 319 high-quality annotated arthropod genomes and uncovered a high-confidence set of 274 HGT-chimeras corresponding to 104 independent origination events across diverse arthropods. HGT-chimeras contain intervals acquired from across the tree of life, and many likely originated via a gene duplication-based mechanism. To assess whether HGT-chimeras might be functionally important, we performed RT-PCR and Sanger sequencing of tissues from 20 arthropod species predicted to harbor HGT-chimeras in their genome. We found evidence for the expression of contiguous chimeric mRNAs for 36 of 41 tested HGT-chimeras across 18 of 20 different tested species. We also found evidence that HGT chimeras evolve under purifying selection and have acquired potentially functional domain architectures, consistent with the hypothesis that these genes are in active use and may participate in diverse biological processes. These results illuminate an underappreciated combinatorial mechanism underlying the origin of novel genes across the largest animal phylum, and suggest that interdomain sequence fusion can play important roles in animal biology and evolution. SignificanceEvolution forges novelty through the repurposing of available parts. Can recently acquired parts, previously foreign to an organism, be similarly repurposed? Applying a rigorous methodology to 319 genomes from arthropods, the largest phylum of animals, we uncover 104 novel genes that arose from the fusion of animal genes with fragments acquired via horizontal gene transfer from bacteria, viruses, fungi, and plants. RNA-Seq and RT-PCR across multiple species show that many of these novel genes are expressed as mRNAs. Many show signatures of evolutionary conservation and coherent domain architectures, suggesting that these chimeric genes may play important roles in diverse biological processes. These results reveal an understudied path to evolutionary innovation via "bricolage" of genes from across the tree of life.

evolutionary biology↗

bioRxiv2025

Computational rheometry of viscoelastic networks: From random graphs to biomolecular condensates

Multivalent biomacromolecules including multi-domain and intrinsically disordered proteins form biomolecular condensates via reversible phase transitions. Condensates are viscoelastic materials that display composition-specific rheological properties and responses to mechanical forces. Graph-based descriptions of microstructures can be combined with computational rheometry to model the outcomes of passive and active mechanical measurements. We consider two types of network models for microstructures. In the Jeffreys model, each edge in the network is a Jeffreys element. In the Stokes-Maxwell model, each edge is a Maxwell element that is embedded in an incompressible viscous fluid that can undergo Stokes flow. We describe results from comparative assessments of the two models for individual elements, ordered lattices, random geometric graphs, structured graphs, and graphs for condensates that are extracted from coarse-grained simulations of disordered proteins. Results from deformation and relaxation tests and flow field analysis reveal how distinct length and time scales contribute to the responses of different types of networks. No single test provides definitive assessments of the connections between material properties and microstructures. Instead, a range of active and passive rheometric tests are essential for distinguishing the responses of different types of networks. Our work establishes computational rheometry as a framework for bridging disparate length and timescales to assess how molecular-scale interactions and dynamics give rise to viscoelastic responses on the mesoscale.

biophysics↗

bioRxiv2025

The PPE2 protein of Mycobacterium tuberculosis is responsible for the development of hyperglycemia and insulin resistance during tuberculosis

Diabetes is a known risk factor for tuberculosis (TB), but clinical evidences suggest that TB itself can induce hyperglycaemia and insulin resistance, though the underlying mycobacterial factors are not known. Herein, we implicate PPE2, a secretory PE/PPE family protein of Mycobacterium tuberculosis (Mtb), as a key modulator of adipose tissue physiology that contributes to the development of insulin resistance. In mice, PPE2 caused fat loss, adipocyte hypertrophy, immune cell infiltration, impaired glucose tolerance, reduced expression of PPAR-{gamma}, C/EBP-, adiponectin and higher insulin resistance. Transcriptomic analysis revealed PPE2 altered expression of genes associated with chemokine/cytokine, ribosomal biogenesis and lipase signaling. PPE2 induced lipolysis by activating cAMP-PKA-HSL axis, increased circulating free fatty acids, a feature also observed in TB patient sera. Interestingly, PPE2-immunization mitigated these effects, suggesting its potential as a subunit vaccine. Overall, this study identifies PPE2 as a key link between Mtb-infection, adipose tissue dysfunction and insulin resistance.

microbiology↗

bioRxiv2025

Mechanistically explainable AI model for predicting synergistic cancer therapy combinations

This study introduces a Large Language Model (LLM)-based framework that combines drug combination data with a knowledge graph to predict synergistic oncology drug combinations with mechanistic insights. Using a retrieval-augmented generation (RAG) approach, over 50,000 in vitro drug pair assay results and 1,631 human clinical trial or preclinical test entries were integrated to enhance predictive accuracy and explainability. Validation achieved an F1 score of 0.80, demonstrating the frameworks potential to streamline drug discovery and improve translational strategies in cancer treatment.

cancer biology↗

bioRxiv2025

HuR-Dependent Cooperative Export of miRNAs From Activated Macrophages

miRNA export is a regulated process vital for maintaining balanced miRNA levels and target gene expression in metazoan cells. RNA-interacting proteins finely adjust the selectivity and specificity of miRNA export, ensuring context-dependent optimal extracellular export of gene-repressive miRNAs in higher eukaryotes. Our findings reveal that activated macrophage cells export miRNAs in a cooperative manner, where elevated expression of miR-122 can significantly enhance the export of miR-146a and selective others. We predict and observe that the cooperative export of repressive miRNAs achieves synchronized upregulation of pro-inflammatory targets in activated macrophage cells. Exploring the molecular basis of this event, we have noted the co-entry of miRNAs into endosomes for their extracellular export, highlighting the selective and cooperative nature of endosome targeting of miRNAs. HuR, a miRNA-binding protein crucial for selectively exporting specific sets of miRNAs from various cell types, facilitates cooperative miRNA entry into endosomes both in the in vivo and in vitro assay conditions. A catalytic amount of high-affinity miR-122 enhances HuRs binding to low-affinity miR-146a, facilitating the efficient endosomal compartmentalization of both miRNAs for their co-export via extracellular vesicles (EVs).

immunology↗

bioRxiv2025

PE11 promotes intracellular persistence of Mycobacterium tuberculosis by inhibiting autophagy and lysosomal biogenesis by targeting the FLCN-lactate-TFEB signaling axis

Mycobacterium tuberculosis (Mtb) employs multiple virulence factors, including cell wall-associated proteins, to evade host immune responses. PE11, a cell wall-localized esterase, contributes to Mtb persistence by facilitating cell wall remodelling and resistance to acidic and antibiotic stress. Herein we describe a novel role of PE11 in subverting host autophagy through disruption of TFEB-mediated lysosomal function. PE11 promotes FLCN-dependent depletion of intracellular lactate to destabilize TFEB and thereby downregulate genes essential for autophagic flux and lysosomal acidification. Using a PE11-deficient Mtb strain, we demonstrate that PE11 targets the FLCN-lactate axis to regulate TFEB stability. Exogenous lactate supplementation restored TFEB stability, enhanced lysosomal acidification, and significantly reduced intracellular bacterial burden. Lactate also synergized with frontline anti-tubercular drugs to improve Mtb clearance. These findings establish PE11 as a key immune evasion factor and highlight lactate as a promising host-directed therapeutic to enhance bacterial killing and reduce antibiotic-associated toxicity.

microbiology↗

bioRxiv2025

Novel polymer fixed-target microfluidic platforms with an ultra-thin moisture barrier for serial macromolecular crystallography

The advent of ultrabright fourth generation X-ray light sources, including X-ray free-electron lasers (XFELs) and diffraction limited synchrotrons, has significantly advanced the field of serial macromolecular protein crystallography (SX). SX experiments demand a continuous supply of fresh microcrystalline sample, ideally while minimizing overall sample consumption. Here, we introduce a novel, robust, and user-friendly polymer film technology that can be assembled in various configurations to encapsulate protein microcrystals and provide sample support for SX. This system provides an efficient hydration barrier over extended durations while maintaining an exceptionally low X-ray background. We have validated this technology by assessing hydration retention under both ambient and ultra-high vacuum conditions, and by evaluating its mechanical stability under XFEL pulses. Furthermore, we have demonstrated the effectiveness of this approach in two room-temperature serial crystallography studies to determine the structure of a 24 kDa Rapid Encystment Phenotype (REP24) protein from Franciscella tularensis.

biophysics↗

bioRxiv2025

From Development to Regeneration: Insights into Flight Muscle Adaptations from Bat Muscle Cell Lines

Skeletal muscle regeneration depends on muscle stem cells, which give rise to myoblasts that drive muscle growth, repair, and maintenance. In bats--the only mammals capable of powered flight--these processes must also sustain contractile performance under extreme mechanical and metabolic stress. However, the cellular and molecular mechanisms underlying bat muscle physiology remain largely unknown. To enable mechanistic investigation of these traits (Graphical Abstract), we established the first myoblast cell lines from the pectoralis muscle of Pteronotus mesoamericanus, a highly maneuverable aerial insectivore. Using both spontaneous immortalization and exogenous hTERT/CDK4 overexpression, we generated two stable cell lines that retain proliferative capacity and differentiate into contractile myotubes. These cells exhibit frequent spontaneous contractions, suggesting robust functional integrity at the neuromuscular junction. In parallel, we performed transcriptomic and metabolic profiling of native pectoralis tissue to define molecular programs supporting muscle specialization. Gene expression analyses revealed enriched pathways for muscle metabolism, development, and regeneration, highlighting the supporting roles in tissue maintenance and repair. Consistent with this profile, the flight muscle is triglyceride-rich, which serves as an important fuel source for energetically demanding processes, including muscle contraction and cellular recovery. Integration of transcriptomic and metabolic data identified three key metabolic modules--glucose utilization, lipid handling, and nutrient signaling--that likely coordinate ATP production and support metabolic flexibility. Together, these complementary tools and datasets provide the first in vitro platform for investigating bat muscle research, enabling direct exploration of muscle regeneration, metabolic resilience, and evolutionary physiology. Graphical AbstractEstablishment of bat muscle cell cultures from the Mesoamerican mustached bat (P. mesoamericnus) provides an in vitro platform to investigate muscle regeneration and flight muscle biology. The pectoralis major muscle was isolated to generate primary myoblast cultures, which were expanded and immortalized using hTERT/CDK4. The resulting myoblast lines retain proliferative and differentiation capacity. RNA sequencing of native pectoralis muscle tissue revealed molecular signatures of myogenic regulation, stress resilience, and tissue remodeling, supporting the relevance of these in vitro models for studying muscle maintenance and regenerative mechanisms. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=127 SRC="FIGDIR/small/662643v1_ufig1.gif" ALT="Figure 1"> View larger version (30K): org.highwire.dtl.DTLVardef@490f49org.highwire.dtl.DTLVardef@8ab615org.highwire.dtl.DTLVardef@b840e1org.highwire.dtl.DTLVardef@ab92eb_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗

bioRxiv2025

Classifier-guided Deep Oscillatory Neural Networks (cDONN) for capturing both Neural Dynamics and Behavior simultaneously

Generating EEG signals alongside behavioural actions introduces substantial biological complexity, akin to an abstract model that mimics rich oscillatory brain dynamics by simultaneously producing dynamic neural activity and corresponding behavioural responses. In this work, we propose a novel class of bio-inspired classifier guided Deep Oscillatory Neural Network (cDONN), designed to simultaneously model and capture both neural signal dynamics and behavioural patterns. The cDONN integrates a feedforward neural network with Hopf oscillator neurons, specialized in handling sequential data generation tasks. The model features a Y-shaped structure with two output branches: one for classifying the input image that signifies behaviour, and the other for generating corresponding EEG signals that represent neural dynamics. We evaluate the performance of the behaviour classification branch using the standard ramp classification accuracy used for the cDONN network. For the EEG signal generation branch, we benchmark the generated signals using classification accuracy via a pre-trained DONN signal classifier, alongside comparison with real EEG signals using metrics such as Inception Score, Frechet Inception Distance (FID), power spectrum analysis and topoplot analysis. Our analysis reveals a strong correspondence between the generated and real EEG data, particularly in the midline parietal region within the alpha frequency band, and in the left hemisphere within the delta band, indicating high fidelity and neurophysiological realism in the synthesized signals. Our approach offers a novel perspective on modelling the simultaneous relationship between neural activity and behaviour, pushing the boundaries of EEG signal generation and behavioural analysis.

bioengineering↗

bioRxiv2025

Evaluating horseradish peroxidase-mimic DNAzyme transducer for glucometer readout: roles of various components and their optimization

The Development of decentralized disease detection elements is integral towards ensuring democratized access to healthcare. Personal glucometers, thanks to their low cost, ease of usage, and universal availability, are increasingly being identified as a central tenet of democratized disease detection. Towards this objective, it will first be critical to establish whether glucometers can be integrated with established molecular diagnosis workflows such as enzyme-linked absorbent assay (ELISA) or its analogous techniques such as enzyme-linked aptasorbent assays (ELASA). In ELISA or its analogous assays, horseradish peroxidase (HRP) or its mimics, such as hemin DNAzyme conjugated to biorecognition elements (e.g., antibody or aptamers), serve as the signal transducers. These transducers generate colorimetric or electrochemical signals measurable in centralized instruments such as ELISA readers or potentiostats. To become compatible with ELISA readouts in decentralized settings, the glucometers should therefore be able to quantify the activity of HRP mimics such as hemin DNAzyme. The current work thus explores the possibility of the use of glucometers as a general readout instrument for hemin DNAzyme transducer activity involving potassium ferrocyanide as a novel redox substrate. Using both absorbance and glucometer readout, we systematically investigated the role of buffer, pH, redox mediator, and H2O2 in achieving optimal readout conditions. It also probed the prospective DNAzyme operational range under these assay conditions. In addition, transferability of optimized assay parameters in other commercial glucometer brands, as well as preliminary suitability in HRP enzyme, was explored. Altogether, our study explores the possibility of using DNAzymes as transducers for glucometers as a democratized alternative to ELISA readers, multimode readers, or potentiostats towards equitable access to disease diagnosis.

biochemistry↗