Comment on, "Single nucleotide polymorphisms as markers of genetic susceptibility for oral potentially malignant disorders risk: Review of evidence to date".

This study by Shridhar et al. 2016 reviews the genetic susceptibility to oral potentially malignant disorders (OPMD) through the analysis of single nucleotide polymorphisms (SNPs). By examining data from 47 studies conducted between 2000 and 2016, the research highlights genetic markers involved in carcinogen metabolism, DNA repair, cell cycle control, and immune-inflammatory responses. Despite the insights provided, the over-reliance on small sample sizes limits the statistical power and generalizability of the findings. Future research should focus on larger, more diverse populations and advanced genotyping technologies to enhance detection of significant genetic variants. Integrating multi-omics data and conducting longitudinal studies will further elucidate the molecular mechanisms underlying OPMD and its progression to oral cancer. Collaborative efforts are essential to validate these findings and develop biomarkers for early detection and prevention.

Comment on, "Characterization of oral microbiota in HPV and non-HPV head and neck squamous cell carcinoma and its association with patient outcomes".

The article "Characterization of oral microbiota in HPV and non-HPV head and neck squamous cell carcinoma and its association with patient outcomes" by Chan et al. investigates the relationship between oral microbiota, HPV infection, and patient outcomes in head and neck squamous cell carcinoma (HNSCC). This comprehensive study, involving 166 Chinese adults, utilized advanced sequencing techniques to profile bacterial and HPV regions in paired tumor and control tissues. The findings highlight the complex interplay between microbiota dysbiosis, HPV infection, and HNSCC progression. Despite the robustness of the study, limitations include potential biases in DNA extraction and PCR amplification, and unaccounted environmental factors. Recommendations for future research include increasing sequencing depth, comparing DNA extraction methods, using multiple bioinformatics pipelines, and controlling for environmental variables. Longitudinal studies and microbiota-targeted interventions are suggested to further elucidate the role of oral microbiota in HNSCC and improve patient outcomes.

Detection and Identification of Various Microplastics in Different Orthodontic Adhesives.

Background Microplastics are acknowledged as significant environmental contaminants. The clinical use of dental materials, particularly adhesives containing plastic polymers, can give rise to the production of plastic micro- and nanoparticles, which subsequently find their way into the environment. The aim of the study was to detect different microplastics and identify them in various orthodontic adhesives. Materials and methods Four different light cure orthodontic adhesives, including Transbond XT (3M Unitek, Monrovia, CA), Ormco Enlight (Ormco, Orange, CA), Orthofix SPA (Orthofix, Verona, Italy), and Aqualine LC (Tomy International Inc, Tokyo, Japan), were collected and placed in separate Eppendorf tubes. Microplastics present in each adhesive were identified using scanning electron microscopy. Subsequently, each specimen was suspended in hydrogen peroxide, placed within a shaking incubator, and analyzed using Fourier transform infrared spectroscopy (FTIR) to identify the type of polymer. Results The scanning electron microscope shows the surface morphology and the most predominant types of microplastics identified were fibers, fragments, and pellets. FTIR results showed the presence of several major functional groups, including hydroxyl, amine, ester, fluoro, and halo groups. Conclusion When contrasted with the quantity of microplastic waste generated by other sectors like the textile, cosmetic, and fishing industries, the microparticulate waste stemming from dental adhesives has a minimal effect on environmental deterioration. Strategies for addressing this concern should give precedence to reducing the use of these materials and adopting effective recovery methods, which could potentially involve recycling processes.

Harnessing the power of mollusc lectins as immuno-protective biomolecules.

The rapid advancement of molecular research on macromolecules has contributed to the discovery of 'Lectin', a carbohydrate-binding protein which specifically interacts with receptors on the surface of glycans and regulates various cellular activities thereby stimulating immunological functions. Considering the wide variety of sources and immunological significance, research has led to the discovery of lectins in invertebrate molluscs. Such lectins in molluscs mediate active immune response as they lack adaptive immunity. Phylum Mollusca is identified with different types of lectins such as C-lectin, Galectin, P-lectin, I-lectin, and H-lectin, along with other immunologically significant lectin molecules such as F- lectin, R-lectin, ficolins, chitinase like lectin etc., all of these with specific ligand binding and structural diversity. Molluscan C-type lectins are the most functional ones that increase the activity of phagocytic cells through specific carbohydrate binding of antigenic ligands and haemocyte adhesion thereby enhancing the immune response. Helix pomatia agglutinin and Helix aspersa agglutinin are the two H-lectins that were identified within molluscs that could even target cancer-progressing cells through specific binding. Also, these lectins identified in molluscs are proven to be efficient in antibacterial and immunomodulatory functions. These insights attract researchers to identify novel lectins in molluscs and their characterization that play a key role in protection against diseases. This review discusses the structural features of mollusc lectins, their specific binding, molecular interactions and their immunological applications.

Random mutagenesis as a tool for industrial strain improvement for enhanced production of antibiotics: a review.

Secondary metabolites are produced by microbes in minimal quantities in the natural environment out of necessity. However, in the pharmaceutical industry, their overproduction becomes essential. To achieve higher yields, genetic modifications are employed to create strains that surpass the productivity of the initially isolated strains. While rational screening and genetic engineering have emerged as valuable practices in recent years, the cost-effective technique of mutagenesis and selection, known as "random screening," remains a preferred method for efficient short-term strain development. This review aims to comprehensively explore all aspects of strain improvement, focusing on why random mutagenesis continues to be widely adopted.

Advancements in Composite Materials and Their Expanding Role in Biomedical Applications.

The synthesis of a Ni-doped ZnO nanocomposite incorporating chitosan (CS/Ni-doped ZnO) was achieved via a precipitation method, followed by annealing at 250 °C. This study comprehensively examined the nanocomposite's structural, functional, morphological, and porosity properties using various analytical techniques, including X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), high-resolution scanning electron microscopy (HR-SEM), transmission electron microscopy (TEM), and Brunauer-Emmett-Teller (BET) analysis. The presence of chitosan (CS) and nickel (Ni) within the nanocomposite, along with their influence on reducing the band gap of ZnO particles and enhancing the generation of electron-hole pairs, was confirmed using UV-visible near-infrared spectroscopy (UV-vis-NIR). The electrochemical properties of the CS/Ni-doped ZnO nanocomposite were investigated via electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) by utilizing a phosphate buffer solution with a pH of 6, which closely resembled the typical pH of bacterial cell walls. Finally, the prepared CS/Ni-doped ZnO nanocomposite was evaluated for its antibacterial and anticancer activities. The results demonstrated the highest inhibition of bacterial growth in P. vulgaris, whereas the lowest inhibition was found in S. aureus across various concentrations, thus highlighting its potential in antimicrobial applications. The cytotoxicity of CS/Ni-doped ZnO nanocomposites demonstrated remarkable effects with a half-maximum inhibitory concentration of approximately 80 ± 0.23 µg mL-1 against MCF-7 breast cancer cell lines, following a dose-dependent manner.

Recent Development and Application of "Nanozyme" Artificial Enzymes-A Review.

Nanozymes represent a category of nano-biomaterial artificial enzymes distinguished by their remarkable catalytic potency, stability, cost-effectiveness, biocompatibility, and degradability. These attributes position them as premier biomaterials with extensive applicability across medical, industrial, technological, and biological domains. Following the discovery of ferromagnetic nanoparticles with peroxidase-mimicking capabilities, extensive research endeavors have been dedicated to advancing nanozyme utilization. Their capacity to emulate the functions of natural enzymes has captivated researchers, prompting in-depth investigations into their attributes and potential applications. This exploration has yielded insights and innovations in various areas, including detection mechanisms, biosensing techniques, and device development. Nanozymes exhibit diverse compositions, sizes, and forms, resembling molecular entities such as proteins and tissue-based glucose. Their rapid impact on the body necessitates a comprehensive understanding of their intricate interplay. As each day witnesses the emergence of novel methodologies and technologies, the integration of nanozymes continues to surge, promising enhanced comprehension in the times ahead. This review centers on the expansive deployment and advancement of nanozyme materials, encompassing biomedical, biotechnological, and environmental contexts.

A Review on the Involvement of Heat Shock Proteins (Extrinsic Chaperones) in Response to Stress Conditions in Aquatic Organisms.

Heat shock proteins (HSPs) encompass both extrinsic chaperones and stress proteins. These proteins, with molecular weights ranging from 14 to 120 kDa, are conserved across all living organisms and are expressed in response to stress. The upregulation of specific genes triggers the synthesis of HSPs, facilitated by the interaction between heat shock factors and gene promoter regions. Notably, HSPs function as chaperones or helper molecules in various cellular processes involving lipids and proteins, and their upregulation is not limited to heat-induced stress but also occurs in response to anoxia, acidosis, hypoxia, toxins, ischemia, protein breakdown, and microbial infection. HSPs play a vital role in regulating protein synthesis in cells. They assist in the folding and assembly of other cellular proteins, primarily through HSP families such as HSP70 and HSP90. Additionally, the process of the folding, translocation, and aggregation of proteins is governed by the dynamic partitioning facilitated by HSPs throughout the cell. Beyond their involvement in protein metabolism, HSPs also exert a significant influence on apoptosis, the immune system, and various characteristics of inflammation. The immunity of aquatic organisms, including shrimp, fish, and shellfish, relies heavily on the development of inflammation, as well as non-specific and specific immune responses to viral and bacterial infections. Recent advancements in aquatic research have demonstrated that the HSP levels in populations of fish, shrimp, and shellfish can be increased through non-traumatic means such as water or oral administration of HSP stimulants, exogenous HSPs, and heat induction. These methods have proven useful in reducing physical stress and trauma, while also facilitating sustainable husbandry practices such as vaccination and transportation, thereby offering health benefits. Hence, the present review discusses the importance of HSPs in different tissues in aquatic organisms (fish, shrimp), and their expression levels during pathogen invasion; this gives new insights into the significance of HSPs in invertebrates.

Exploring the Antimicrobial Potential and Biofilm Inhibitory Properties of Hemocyanin from Hemifusus pugilinus (Born, 1778).

The seafood industry plays a huge role in the blue economy, exploiting the advantage of the enriched protein content of marine organisms such as shrimps and molluscs, which are cultured in aquafarms. Diseases greatly affect these aquatic organisms in culture and, hence, there is need to study, in detail, their innate immune mechanisms. Hemocyanin is a non-specific innate defense molecule present in the blood cells of several invertebrates, especially molluscs, arthropods, and annelids. It is concerned with oxygen transport, blood clotting, and immune enhancement. In the present study, this macromolecular metalloprotein was isolated from the hemolymph of the marine snail Hemifusus pugilinus (Born, 1778) using Sephadex G-100 gel filtration column chromatography. It occurred as a single band (MW 80 kDa) on SDS-PAGE. High-performance liquid chromatography (HPLC) of the purified hemocyanin showed a single peak with a retention time of 4.3 min. The secondary structure and stability of the protein were detected using circular dichroism (CD), and the spectra demonstrated negative ellipticity bands close to 208 nm and 225 nm, indicating ß-sheets. Further exploration of the purified hemocyanin revealed remarkable antimicrobial and antibiofilm activities against Gram-positive (Enterococcus faecalis and Staphylococcus aureus) and Gram-negative bacteria (Pseudomonas aeruginosa and Proteus vulgaris) at a concentration of 1-5 µg/mL. Spectrophotometric and in situ microscopic analyses (CLSM) unveiled the potential of the purified hemocyanin to inhibit biofilm formation in these bacteria with a minimal inhibitory concentration of 40 µg/mL. Furthermore, H. pugilinus hemocyanin (10 µg/mL concentration) displayed antifungal activity against Aspergillus niger. The purified hemocyanin was also assessed for cytotoxicity against human cancer cells using cell viability assays. Altogether, the present study shows that molluscan hemocyanin is a potential antimicrobial, antibiofilm, antifungal, anticancer, and immunomodulatory agent, with great scope for application in the enhancement of the immune system of molluscs, thereby facilitating their aquaculture.

Control and prevention of microbially influenced corrosion using cephalopod chitosan and its derivatives: A review.

Microbially influenced corrosion (MIC) of metals is an important industrial problem, causing 300-500 billion dollars of economic loss worldwide each year. It is very challenging to prevent or control the MIC in the marine environment. Eco-friendly coatings embedded with corrosion inhibitors developed from natural products may be a successful approach for MIC prevention or control. As a natural renewable resource, cephalopod chitosan has a number of unique biological properties, such as antibacterial, antifungal and non-toxicity effects, which attract scientific and industrial interests for potential applications. Chitosan is a positively charged molecule, and the negatively charged bacterial cell wall is the target of its antimicrobial action. Chitosan binds to the bacterial cell wall and disrupts the normal functions of the membrane by, for example, facilitating the leakage of intracellular components and impeding the transport of nutrients into the cells. Interestingly, chitosan is an excellent film-forming polymer. Chitosan may be applied as an antimicrobial coating substance for the prevention or control of MIC. Furthermore, the antimicrobial chitosan coating can serve as a basal matrix, in which other antimicrobial or anticorrosive substances like chitosan nanoparticles, chitosan silver nanoparticles, quorum sensing inhibitors (QSI) or the combination of these compounds, can be embedded to achieve synergistic anticorrosive effects. A combination of field and laboratory experiments will be conducted to test this hypothesis for preventing or controlling MIC in the marine environment. Thus, the proposed review will identify new eco-friendly MIC inhibitors and will assay their potential in future applications in the anti-corrosion industry.

Innate Immune Response Assessment in Cyprinus carpio L. upon Experimental Administration with Artemia salina Bio-Encapsulated Aeromonas hydrophila Bacterin.

The present study aimed to analyze the enhancement of innate immune responses in juvenile-stage common carp (Cyprinus carpio L.), upon the administration of heat-killed Aeromonas hydrophila at a dosage of 1 × 107 CFU ml-1 through bio-encapsulation in the aquatic crustacean, Artemia salina. This work emphasizes the modulation of innate immune response when administered with the bio-encapsulated heat-killed antigen that acts as an inactivated vaccine against Motile Aeromonas Septicemia disease. Bio-encapsulated oral administration of antigens promotes innate immunity in juvenile-stage fishes. The optimization of effective bio-encapsulation of bacterin in Artemia salina nauplii was carried out and the best optimal conditions were chosen for immunization. The functional immune parameters such as myeloperoxidase, lysozyme, alkaline phosphatase, antiprotease and respiratory burst activity in serum, blood and intestinal tissue samples were analyzed along with blood differential leukocyte count and tissue histopathology studies. Both humoral and cellular immune responses analyzed were substantially induced or enhanced in the treatment groups in comparison with the control group. The results showed a significant variation in the bio-encapsulation group than the control group and also were comparable to the protection conferred with immersion route immunization under similar conditions. Thus, most of the innate non-specific immune responses are inducible, despite being constitutive of the fish immune system, to exhibit a basal level of protection and a road to better vaccination strategy in Cyprinus carpio L. aquaculture worldwide.

Back2Basics: animal lectins: an insight into a highly versatile recognition protein.

The rapid advancement of molecular research has contributed to the discovery of 'Lectin', a carbohydrate-binding protein which specifically interacts with receptors on surface glycan moieties that regulate various critical cellular activities. The first animal lectin reported was 'the asialoglycoprotein receptor' in mammalian cells which helped analyze how animal lectins differ in glycoconjugate binding. Animal lectins are classified into several families, depending on their diverse cellular localization, and the binding specificities of their Carbohydrate-Recognition Domain (CRD) modules. Earlier characterization of animal lectins classified them into two structural families, the C-type (Ca2+-dependent binding) and S-type galectins (sulfhydryl-dependent binding) lectins. The C-type lectin includes the most significant animal lectins, such as endocytic receptors, mannose receptors, selectins, and collectins. The recent developments in research based on the complexity of the carbohydrate ligands, the metabolic processes they perform, their expression levels, and their reliance on divalent cations have identified more than 100 animal lectins and classified them into around 13 different families, such as Calnexin, F-lectin, Intelectin, Chitinase-like lectin, F-box lectin, etc. Understanding their structure and expression patterns have aided in defining their significant functions including cell adhesion, antimicrobial activity, innate immunity, disease diagnostic biomarkers, and drug delivery through specific carbohydrate-protein interactions. Such extensive potential roles of animal lectins made it equally important to plant lectins among researchers. Hence, the review focuses on providing an overview of animal lectins, their taxonomy, structural characteristics, and functions in diverse aspects interconnected to their specific carbohydrate and glycoconjugate binding.

Oral vaccination for sustainable disease prevention in aquaculture-an encapsulation approach.

The prevalence of infectious diseases in the aquaculture industry and a limited number of safe and effective oral vaccines has imposed a challenge not only for fish immunity but also a threat to human health. The availability of fish oral vaccines has expanded recently, but little is known about how well they work and how they affect the immune system. The unsatisfactory efficacy of existing oral vaccinations is partly attributable to the antigen degradation in the adverse gastrointestinal environment of fishes, the highly tolerogenic gut environment, and inferior vaccine formulation. To overcome such challenges in designing: an easier, cost-efficient, and effective vaccination method, several encapsulation methods are being adopted to safeguard antigens from the intestinal atmosphere for their immunogenic functions. Oral vaccination is easily degraded by gastric acids and enzymes before reaching the immunological site; however, this issue can be solved by encapsulating antigens in poly-biodegradable nanoparticles, transgenic designed bacteria, plant systems, and live feeds. To enhance the immunological impact, each antigen delivery method operates at a different level. Utilizing nanotechnology, it has been possible to regulate vaccination parameters, target particular cells, and lower the antigen dosage with potent nanomaterials such as chitosan, poly D,L-lactic-co-glycolic acid (PLGA) as vaccine carriers. Live feeds such as Artemia salina can be utilized as bio-carrier, owing to their appropriate size and non-filter feed system, through a process called bio-encapsulation. It ensures the protection of antigens over the fish intestine and ensures complete uptake by immune cells in the hindgut for increased immune response. This review comprises recent advances in oral vaccination in aquaculture in terms of an encapsulation approach that can aid in future research.

Green Synthesis and Characterization of Xanthium strumarium-Mediated Titanium Dioxide Nanoparticles.

Background Green synthesis of nanoparticles is a growing trend. The annual plant Xanthium strumarium L. (X. strumarium) belongs to the Asteraceae family. The herb has traditionally been used to treat a variety of ailments, including leucoderma, dangerous insect bites, epilepsy, salivation, allergic rhinitis, sinusitis, etc. Inorganic, biocompatible, and non-toxic titanium is a substance employed in the pharmaceutical and biomedical industries as well as in fields like bone tissue engineering. The aim of the study is to characterize titanium dioxide nanoparticles (TiO2NPs), which were synthesized from X.strumarium. Also, this study aims to assess the cytotoxic properties of the synthesized leaf extract and the TiO2NPs. Materials and methods In this study, the biosynthesis of TiO2NPs was made from X. strumarium leaf extract. The characterization of the green-synthesized TiO2NPs was done using the spectral analysis of an ultraviolet (UV)-visible spectrophotometer, scanning electron microscopy (SEM), and Fourier Transform Infrared Spectroscopy (FTIR). The advantage of using TiO2NPs is that they possess antimicrobial, antibacterial, chemical stability, and catalytic properties. The leaf extract and the biosynthesized nanoparticles were tested against human fibroblast cell lines for biocompatibility using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay.  Results SEM investigation showed that TiO2NPs were crystalline in nature. FTIR confirms the presence of alkyne and amine functional groups, and the pointed vertices in the X-ray diffraction (XRD) pattern show the crystalline nature of TiO2NPs. The study found that the cell viability of TiO2NPs was 110%. Conclusion TiO2NPs were synthesized from X. strumarium leaf extract and characterized using SEM, FTIR, and XRD. The TiO2NPs were found to be crystalline in nature with various functional groups. MTT assay shows that the synthesized nanoparticles are promising biocompatible agents that can be used in future research in the medical field.

Morphological and functional characterization of circulating hemocytes using microscopy techniques.

In the present study, Microscopy studies were performed to characterize the blood cells of the mangrove crab Episesarma tetragonum. Three types of hemocytes were observed: granulocytes, semi-granulocytes, and hyalinocytes or agranulocytes. Hyalinocytes have a distinguished nucleus surrounded by the cytoplasm, and a peculiar cell type was present throughout the cytosol, lysosomes with hemocyte types (granules) stained red (pink). Giemsa staining was used to differentiate between the large and small hemocytes. Ehrlich's staining was used to differentiate granule-containing cells in acidophils (55%), basophils (44%), and neutrophils (<1%). Periodic acid-Schiff staining was used to identify the sugar molecules in the cytoplasm. Cell-mediated immune reactions including phagocytosis, encapsulation, agglutination, and peroxidase-mediated cell adhesion are the functions of hemocytes. Agglutination reaction involves both kind of cells involved in yeast and heme-agglutination responses in invertebrates. The beta glucan outer layer of yeast cells was recognized by hemocyte receptors. Human RBC cells were agglutinated via granulocytes. E. tetragonum hemocytes are an important animal model for studying both ultrastructural and functional activity of circulating cells. In addition, E. tetragonum hemocytes exhibited excellent antibacterial and antibiofilm activities were studied through plating and microplate assays. Biofilm inhibition was also visualized through changes in biochemical assays and morphological variations were visualized through levels in in situ microscopy analysis.

In-vitro dissolution and microbial inhibition studies on anticancer drug etoposide with ß-cyclodextrin.

In this article, we have reported the inclusion complex behaviors and their pharmaceutical application of anticancer drug property of Etoposide with ß-cyclodextrin. The inclusion complex is used to improve the poor aqueous solubility of the anticancer drug Etoposide. The aqueous solubility and in-vitro dissolution studies support to the anticancer drug Etoposide with ß-cyclodextrin complex is significantly improves the aqueous solubility. Etoposide:ß-cyclodextrin solid-state complexes were prepared by Physical mixture, kneading and solvent evaporation methods, and were characterized by FT-IR, 1HNMR, XRD, DSC and SEM techniques. In addition, the in-vitro antimicrobial and antibiofilm study of Etoposide drug is a sensible microorganism was significantly increased by an inclusion complexation process. The antibiofilm of anticancer drug Etoposide with ß-cyclodextrin studies have been analyzed by confocal laser scanning microscopy.

Selective recognition and stabilization of new ligands targeting the potassium form of the human telomeric G-quadruplex DNA.

The development of a ligand that is capable of distinguishing among the wide variety of G-quadruplex structures and targeting telomeres to treat cancer is particularly challenging. In this study, the ability of two anthraquinone telomerase inhibitors (NSC749235 and NSC764638) to target telomeric G-quadruplex DNA was probed. We found that these ligands specifically target the potassium form of telomeric G-quadruplex DNA over the DNA counterpart. The characteristic interaction with the telomeric G-quadruplex DNA and the anticancer activities of these ligands were also explored. The results of this present work emphasize our understanding of the binding selectivity of anthraquinone derivatives to G-quadruplex DNA and assists in future drug development for G-quadruplex-specific ligands.

Tamarind seed coat ameliorates fluoride induced cytotoxicity, oxidative stress, mitochondrial dysfunction and apoptosis in A549 cells.

Fluoride (F) is an environmental contaminant and industrial pollutant. Molecular mechanisms remain unclear in F induced pulmonary toxicity even after numerous studies. Tamarind fruits act as defluoridating agents, but no study was conducted in in vitro systems. Hence, we aimed to assess the ameliorative impact of the tamarind seed coat extract (TSCE) against F toxicity utilizing lung epithelial cells, A549. Cells were exposed to sodium fluoride (NaF-5 mM) alone and in combination with TSCE (750 ng/ml) or Vitamin C (positive control) for 24 h and analyzed for F content, intracellular calcium ([Ca(2+)]i) level, oxidative stress, mitochondrial integrity and apoptotic markers. TSCE treatment prevented the F induced alterations in [Ca(2+)]i overload, F content, oxidant (reactive oxygen species generation, lipid peroxidation, protein carbonyl content and nitric oxide) and antioxidant (superoxide dismutase, catalase, glutathione peroxidase and glutathione) parameters. Further, TSCE modulates F activated changes in mitochondrial membrane potential, permeability transition pore opening, cytochrome-C release, Bax/Bcl-2 ratio, caspase-3 and PARP-1 expressions. In conclusion, our study demonstrated that TSCE as a potential protective agent against F toxicity, which can be utilized as a neutraceutical.

Structure-based virtual screening and experimental validation of the discovery of inhibitors targeted towards the human coronavirus nucleocapsid protein.

Nucleocapsid protein (NP), an essential RNA-binding viral protein in human coronavirus (CoV)-infected cells, is required for the replication and transcription of viral RNA. Recent studies suggested that human CoV NP is a valid target for antiviral drug development. Based on this aspect, structure-based virtual screening targeting nucleocapsid protein (NP) was performed to identify good chemical starting points for medicinal chemistry. The present study utilized structure-based virtual screening against human CoV-OC43 using the Zinc database, which is performed through docking with varying precisions and computational intensities to identify eight potential compounds. The chosen potential leads were further validated experimentally using biophysical means. Surface plasmon resonance (SPR) analysis indicated that one among the potential leads, 6-chloro-7-(2-morpholin-4-yl-ethylamino) quinoxaline-5,8-dione (small-compound H3), exhibited a significant decrease of RNA-binding capacity of NP by more than 20%. The loss of binding activity was manifested as a 20% decrease in the minimum on-rate accompanied with a 70% increase in the maximum off-rate. Fluorescence titration and X-ray crystallography studies indicated that H3 antagonizes the binding between HCoV-OC43 NP and RNA by interacting with the N-terminal domain of the NP. Our findings provide insight into the development of new therapeutics that disrupt the interaction between RNA and viral NP in the HCoV. The discovery of the new compound would be an impetus to design novel NP inhibitors against human CoV.