Search Results (202)

Bacterial toxins, such as Staphylococcal Enterotoxin B (SEB) and Shiga Toxins (STX1, STX2), pose severe public health risks and significant biological threats, demanding rapid and precise qPCR detection. This study reports the initial stages of standardization for uniplex qPCR components, serving as a foundational step toward a future multiplex detection system. The annealing temperature was successfully optimized for the stx1, seb, and 16S rRNA targets, showing high consistency in Cq values and fluorescence intensities at 58.4 °C. The optimized primer ratios ensure maximum amplification efficiency while minimizing potential molecular competition. These optimized assays provide a robust foundation for the multiplex qPCR platform. Once fully validated, this protocol will enhance timely and effective responses, as well as improve preparedness and readiness within the Brazilian biodefense system. Full article

Francisella tularensis, the causative agent of tularemia, is a highly infectious Category A biothreat agent characterized by an exceptionally low infectious dose and diverse transmission routes. Due to the pathogen’s fastidious growth requirements and the high risk of laboratory-acquired infections, traditional cultivation methods are often protracted and hazardous. Consequently, the development of rapid and sensitive diagnostic tools is paramount. This manuscript provides a comprehensive overview of the current landscape of immunoassays, with a specific focus on the evolution from standard laboratory techniques to advanced biosensors. We detail the critical phases of antigen preparation, including high-pressure homogenization and sonication, and the generation of high-affinity polyclonal and monoclonal antibodies. Furthermore, we evaluate the implementation of novel biosensor-like devices, such as electrochemiluminescence and Surface-Enhanced Raman Scattering platforms, designed for point-of-care and field-ready scenarios. By synthesizing recent advancements in nanomaterial-enhanced recognition and microfluidic integration, this review emphasizes the pivotal role of these technologies in achieving early detection and mitigating the impact of both natural outbreaks and potential deliberate misuse of F. tularensis. Full article

Yersinia pestis and Francisella tularensis are Tier-1 pathogens with high interest for biodefense and public health. Evaluating the antibacterial activity of repurposed drugs against these high-priority pathogens is a key element in the ongoing effort to develop diversified antimicrobial strategies. Drug repurposing offers a cost-effective and time-efficient approach to address antibiotic resistance by identifying new applications for existing therapeutics. In this study, we demonstrate in vitro antibacterial effect of the antifungal agent ciclopirox and offer this drug as a potential antibacterial treatment. Ciclopirox in vitro activity was previously reported against various Gram-negative bacteria, including resistant strains, primarily through iron chelation that disrupts key metabolic pathways and virulence mechanisms. Additionally, it exhibits antibiofilm activity and can potentiate the efficacy of certain antibiotics. Our findings reveal that ciclopirox effectively inhibits the in vitro growth of fully virulent strains of Y. pestis and F. tularensis, as well as avirulent isolates, including avirulent mutants that their wild-type susceptibility was reduced through selection to MIC levels defining them as “nonsusceptible” to ciprofloxacin (Y. pestis Kim53Δ70Δ10 and F. tularensis LVS) and doxycycline (LVS), or resistant to doxycycline (Kim53Δ70Δ10) according to CLSI interpretive criteria. Additionally, prolonged exposure of Y. pestis and F. tularensis to sub-MIC and MIC concentrations of ciclopirox did not lead to an increase in observed MIC during the study period. These results highlight ciclopirox as a potential candidate for treatment alternative, combined with other antibiotic substances or repurposed drugs against these bacterial threats. Full article

Cancer immunoprevention leverages the immune system’s surveillance mechanisms to mitigate tumor development. Vaccines that constitute a tumor antigen and an immune adjuvant are perceived as immunoprevention modalities. However, relevant tumor antigens are unknown for non-viral cancers, which constitute most human cancers. Our group has recently shown that SA–4–1BBL, a novel agonist of CD137 receptor, but not antibodies, shows immunoprevention efficacy against various tumors. Advanced bioinformatics analyses of bulk RNA-seq data were conducted to elucidate mechanisms underlying cancer immunoprevention. Mice received subcutaneous injections of SA–4–1BBL or agonistic 3H3 antibody, and the injection-site tissue (IS) and draining lymph nodes (LN) were analyzed for differential gene expression. SA–4–1BBL induced a compartmentalized and temporally dynamic immune program characterized by early effector activation at IS and sustained immune regulation in draining LN. K-means clustering of 4564 DEGs identified eight functionally distinct clusters. IS-enriched clusters contained activation genes for CD4⁺ T and NK cells, including Cd28, Klra1, Cd4, Cd40, and Cd40l, while LN clusters were enriched for regulatory genes (Tnfaip3, Irf5, Col1a2) that ensure immune priming and homeostatic restraint for a balanced response. SA–4–1BBL generated a more selective and durable activation of adaptive immunity, TCR signaling, Th1/Th2 differentiation, and NK cytotoxicity. 3H3 activated broader innate inflammatory programs, including Toll-like receptor and neurodegeneration-linked pathways. IMPRes analysis showed that SA–4–1BBL activates sequential immune-regulatory circuits centered on Stat1, Cd247, and Ifng and modulates the CD151–TGF-β axis. These findings demonstrate that SA–4–1BBL elicits a balanced immune response, ensuring both safety and efficacy in preventing cancer development. Full article

This systematic review article provides a comprehensive and critical analysis of the use of bioinputs in sustainable agriculture, focusing on biosurfactants and absorbent polymers, particularly bacterial cellulose. The article contextualises the growing challenges in agricultural production due to population growth, climate change, and environmental limitations, highlighting the need for alternatives to traditional synthetic inputs that exert negative environmental impacts. The article details functions, types, and benefits, emphasising the ability of bioinputs to improve soil fertility, increase the efficiency of nutrient use, enhance plant resistance to biotic and abiotic stress, and reduce the ecological footprint of agriculture. Emerging biotechnologies are discussed, such as the combined use of biosurfactants with natural polymers to ensure sustainability and efficiency. This article offers an updated description of recent scientific and technological evidence and addresses the potential and limitations of these biological inputs in the global context of modern agriculture. Full article

Background: Venezuelan equine encephalitis virus (VEEV) poses a significant public health and biodefense threat due to periodic epidemics of severe neurological disease in the Americas, yet no licensed human vaccines or specific antiviral therapies exist. Methods: We comprehensively reviewed the current literature across three core domains: structural biology, vaccine development, and therapeutic antibodies. Results: First, we detail the complex viral structural proteome (including E1/E2 glycoproteins and Capsid), focusing on the LDLRAD3 entry receptor interaction. Second, we overview vaccine strategies, covering live-attenuated, VLP, and nucleic acid platforms designed to induce robust neutralizing antibodies. Finally, we examine the evolution of therapeutic antibodies, highlighting that optimal protection often relies on both neutralization and Fc effector functions, particularly for antibodies targeting the fusion loop or receptor-binding sites. Conclusions: Integrating structural insights with advanced antibody and vaccine engineering is essential for establishing effective clinical interventions capable of preventing future outbreaks and treating infected individuals. Full article

The convergence of artificial intelligence and synthetic biology is innovating and accelerating the design of novel viral genomes, expanding both therapeutic opportunities and dual-use risk. This review articulates a countermeasure strategy for emerging and engineered viruses leveraging the programmable CRISPR modality. Building on mounting in vitro and in vivo evidence that Cas9 degrades DNA viruses (e.g., Orthopoxviruses, HSV-1, ASFV), while Cas13 targets RNA viral genomes (e.g., Influenza A, Dengue, RSV), both leading to reduced viremia, diminished disease burden, and alleviated symptoms. Here, we outline a rapid-response pipeline to position CRISPR-based countermeasures in translational and pandemic-response frameworks, linking real-time sequencing to AI-assisted gRNA selection and multiplexed cassette design to achieve viral targeting efficacy. To minimize resistance and off-target risk, we emphasize multi-gRNA cocktails, continuous genomic surveillance, and adaptive gRNA rotation. We also propose governance mechanisms, such as pre-cleared gRNA repositories, transparent design logs, standardized off-target/safety screening, and alignment with evolving nucleic-acid-synthesis screening frameworks to enable emergency deployment while preserving security. Furthermore, compressing the time from sequence to treatment and complementary to vaccines and small-molecule antivirals, CRISPR represents a technologically agile and strategically essential capability to combat both natural outbreaks and AI-enabled biothreats. Collectively, programmable CRISPR antivirals represent an auditable, rapidly adaptable foundation for next-generation biodefense preparedness. Full article

The growing threat of airborne biological agents necessitates rapid, sensitive, and portable detection systems to mitigate risks to public health and national security. We present a comprehensive overview of biosensor technologies developed for airborne biothreat detection, with a focus on aptamer-based electrochemical sensors. These sensors offer key advantages in portability, chemical stability, and adaptability for multiplexed detection in field settings. The urgency for real-time surveillance tools capable of identifying viral, bacterial, and toxin-based agents is discussed, particularly in the context of biodefense. Aerosolized particle capture strategies are reviewed, focusing on microfluidics for micron-sized particles and condensation-based systems for submicron-sized particles, which are preferred for their small-volume operation and seamless integration with biosensors. Key biosensor components are described, including recognition elements—such as aptamers—and transduction mechanisms like electrochemical impedance spectroscopy. EIS is highlighted for its label-free, miniaturizable, and real-time readout capabilities, making it well-suited for portable biosensors. Advances in sensing strategies for both viral and bacterial targets are explored, featuring innovations in nanoporous membrane platforms, nanomaterials, and multiplexed assay formats. Recent developments demonstrate improved sensitivity through nanopore-based signal amplification and enhanced selectivity using engineered aptamer libraries. The review concludes by addressing current limitations, including environmental stability, system integration, and the need for validation with complex real-world samples. Future directions point toward the development of fully integrated, field-deployable biosensing platforms that combine effective aerosol capture with robust and selective biosensing technologies. Full article

Historically, saxitoxin (STX) and its analogs, like neosaxitoxin (NeoSTX), have been restricted to scientific research due to their potent toxicity, despite their potential as therapeutic agents. This study aimed to evaluate the pharmacological versatility of STX and NeoSTX through molecular docking. Using the SwissTargetPrediction server, 18 potential human therapeutic targets were identified, with the 5-HT6 receptor being the primary target. Molecular docking assays were performed with AutoDock Vina version 1.1.2, and the lowest-energy pose for NeoSTX (−5.7 kcal/mol) suggested a stronger binding affinity compared to STX (−5.5 kcal/mol). The analysis of the binding mode revealed key interactions, including hydrogen bonds with Thr199 and hydrophobic contacts with Phe285. These findings provide robust hypotheses regarding the therapeutic potential of NeoSTX as a 5-HT6 receptor antagonist, supporting the development of novel bioactive agents derived from these marine toxins. Full article

Background: The rapid and reliable identification of unknown or highly variable biological toxin proteins, such as the potent Ricin toxin, remains a critical challenge in biodefense and public security. Methods: To address this, we developed a Heuristic De Novo Sequencing (HDPS) strategy, which combines multiple enzymatic and microwave-assisted acid hydrolysis to generate diverse peptides, followed by a two-stage assembly process integrating de novo sequencing with homology-based database searching for robust error correction. Results: When applied to Ricin, this approach achieved 100% sequence coverage for both its A and B chains, with amino acid-level accuracies of 98.13% and 98.47%, respectively, and successfully corrected potential sequencing ambiguities. Conclusions: These results demonstrate that HDPS is a highly accurate and effective method for the de novo sequencing of full-length proteins, making it particularly valuable for characterizing unknown or mutated toxins in the absence of comprehensive reference databases. Full article

Plant-derived toxins such as ricin and abrin represent some of the most potent biological agents known, posing significant threats to public health and security due to their high toxicity, relative ease of extraction, and widespread availability. These ribosome-inactivating proteins (RIPs) have been implicated in politically and criminally motivated events, underscoring their critical importance in the context of biodefense. Public safety agencies, including law enforcement, customs, and emergency response units, require rapid, sensitive, and portable detection methods to effectively counteract these threats. However, many existing screening technologies lack the capability to detect biotoxins unless specifically designed for this purpose, revealing a critical gap in current biodefense preparedness. Consequently, there is an urgent need for robust, field-deployable detection platforms that operate reliably under real-world conditions. End-users in the security and public health sectors demand analytical tools that combine high specificity and sensitivity with operational ease and adaptability. This review provides a comprehensive overview of the biochemical characteristics of ricin and abrin, their documented misuse, and the challenges associated with their detection. Furthermore, it critically assesses key detection platforms—including immunoassays, mass spectrometry, biosensors, and lateral flow assays—focusing on their applicability in operational environments. Advancing detection capabilities within frontline services is imperative for effective prevention, timely intervention, and the strengthening of biosecurity measures. Full article

Optimal sensing devices exhibit a combination of key performance attributes, including an extensive detection limit, exceptional selectivity, high sensitivity, consistent repeatability, precise measurement, and rapid response times with efficient analyte flow. In recent years, biosensing platforms incorporating nanoscale materials have garnered considerable attention due to their diverse applications across various scientific and technological domains. The integration of nanoparticles (NPs) in biosensor design primarily bridges the dimensional gap between the signal transduction element and the biological recognition component, both of which operate at nanometer scales. The synergistic combination of NPs with electrochemical techniques has facilitated the development of biosensors characterized by enhanced sensitivity and superior analyte discrimination capabilities. This comprehensive analysis examines the evolution and recent advancements in nanomaterial (NM)-based biosensors, encompassing an extensive array of nanostructures. These consists of one-dimensional nanostructures including carbon nanotubes (CNTs), nanowires (NWs), nanorods (NRs), and quantum dots (QDs), as well as noble metal and metal and metal oxide nanoparticles (NPs). The article examines how advancements in biosensing techniques across a range of applications have been fueled by the growth of nanotechnology. Researchers have significantly improved biosensor performance parameters by utilizing the distinct physiochemical properties of these NMs. The developments have increased the potential uses of nanobiosensors in a wide range of fields, from food safety and biodefense to medical diagnostics and environmental monitoring. The continuous developments in NM-based biosensors are the result of the integration of several scientific areas, such as analytical chemistry, materials science, and biotechnology. This interdisciplinary approach continues to drive innovations in sensor design, signal amplification strategies, and data analysis techniques, ultimately leading to more sophisticated and capable biosensing platforms. As the field progresses, challenges related to the scalability, reproducibility, and long-term stability of nanobiosensors are being addressed through innovative fabrication methods and surface modification techniques. These efforts aim to translate the promising results observed in laboratory settings into practical, commercially viable biosensing devices that can address real-world analytical challenges across various sectors. Full article

Since the emergence of SARS-CoV-2, the interplay of human behavior, environmental factors, viral evolution, and public health interventions has resulted in unexpected changes in the timing, intensity, and geography of respiratory virus outbreaks. For example, respiratory syncytial viruses (RSV) exhibited a surge during atypical summer months in several countries. Influenza, on the other hand, nearly vanished in the early years of the pandemic, but returned with unusual strength and altered seasonal patterns. Concurrently, new variants of concern in coronaviruses have demonstrated increased airborne transmissibility, greater resilience to environmental conditions, and the ability to evade both natural and vaccine-induced immunity. In this review article, we have synthesized the current understanding of the aerobiology of respiratory infectious viruses, with a particular emphasis on the paradoxical trends observed in recent years. We examined various aspects, including viral morphology and environmental survivability, shifts in seasonality, the drivers of mutation and resistance, and the impact of environmental and climatic factors. Key issues we explored include viral morphology adaptation in response to airborne selective pressures and climate variability influence on the ecology of airborne viruses. Lastly, we investigated future risks and proposed an interdisciplinary framework for monitoring and mitigating airborne viral threats in an ever-changing world. Full article

Fasciola hepatica remains a global health and economic concern, and treatment still relies heavily on triclabendazole. At the parasite–host interface, F. hepatica calcium-binding proteins (FhCaBPs) have a unique EF-hand/DLC-like domain fusion found only in trematodes. This makes it a parasite-specific target for small compounds and vaccinations. To enable novel therapeutic strategies, we report the first elevated-resolution structure of a full-length FhCaBP4. The apo structure was determined at 1.93 Å resolution, revealing a homodimer architecture that integrates an N-terminal, calmodulin-like, EF-hand pair with a C-terminal dynein light chain (DLC)-like domain. Structure-guided in silico mutagenesis identified a flexible, 16-residue β4–β5 loop (LTGSYWMKFSHEPFMS) with an FSHEPF core that demonstrates greater energetic variability than its FhCaBP2 counterpart, likely explaining the distinct ligand-binding profiles of these paralogs. Molecular dynamics simulations and AlphaFold3 modeling suggest that EF-hand 2 acts as the primary calcium-binding site, with calcium coordination inducing partial rigidification and modest expansion of the protein structure. Microscale thermophoresis confirmed calcium as the major ligand, while calmodulin antagonists bound with lower affinity and praziquantel demonstrated no interaction. Thermal shift assays revealed calcium-dependent stabilization and a merger of biphasic unfolding transitions. These results suggest that FhCaBP4 functions as a calcium-responsive signaling hub, with an allosterically coupled EF-hand–DLC interface that could serve as a structurally tractable platform for drug targeting in trematodes. Full article