Salvatore Giovanni De Simone

The Americas encompass an exceptional diversity of terrestrial, freshwater, and marine ecosystems, ranging from Arctic and boreal environments to temperate forests, deserts, tropical rainforests, savannas, wetlands, high-altitude ecosystems, and highly productive coastal and deep-sea environments. This ecological diversity has generated an equally remarkable chemical diversity of specialized metabolites produced by plants, fungi, bacteria, marine organisms, and plant-associated microorganisms. Natural products chemistry in the Americas has historically contributed important therapeutic agents and chemical prototypes, including quinine from Cinchona species, emetine from Carapichea ipecacuanha, paclitaxel from Taxus brevifolia, podophyllotoxin from Podophyllum species, and marine-derived nucleoside analogs that inspired cytarabine. Despite this scientific legacy, the enormous chemical potential of American biodiversity remains incompletely explored. In recent decades, natural products research in the Americas has evolved from conventional extraction, bioactivity-guided fractionation, and structure elucidation toward integrated workflows combining metabolomics, high-resolution mass spectrometry, nuclear magnetic resonance spectroscopy, molecular networking, genome mining, metagenomics, synthetic biology, chemoinformatic, artificial intelligence, and advanced bioassays. These approaches are particularly relevant to biodiverse regions of Latin America and the Caribbean, where traditional medicinal knowledge provides an important source of hypotheses for chemical investigation. At the same time, natural product research must increasingly incorporate biodiversity conservation, equitable benefit sharing, Indigenous rights, sustainable collection, reproducibility, and regulatory compliance. This review discusses the historical foundations, major ecosystems and chemical resources, representative plant, microbial, and marine natural products, methodological advances, translational challenges, and future opportunities in natural products chemistry across the Americas. We pay particular attention to Latin American biodiversity, where a substantial gap persists between biological richness and the number of natural products successfully translated into standardized medicines. We propose that the next generation of natural products research in the Americas should integrate biodiversity science, traditional knowledge, advanced analytical chemistry, computational biology, synthetic biology, and sustainable biomanufacturing to turn chemical diversity into scientifically validated, socially responsible innovation.

Vaccination remains one of the most successful interventions in medical history

Lyme borreliosis (LB) is a tick-borne disease caused by a diverse and expanding group of spirochetes characterized by complex biology and advanced immune evasion mechanisms. It presents a wide range of clinical symptoms affecting multiple organ systems and can lead to persistent complications. The pathogenesis of LB remains incompletely understood, and diagnosis typically relies on serologic assays to detect antibodies against LB. However, the standard two-tiered testing (STTT) algorithm is limited by cross-reactivity, low sensitivity, and delayed results, hindering timely and accurate diagnosis. Although molecular tests are considered the gold standard, their reliance on centralized laboratories can delay critical treatment decisions. This underscores the urgent need for rapid, reliable diagnostic tools, particularly for use at the point of hospital admission. Point-of-care serological and direct antigen testing can provide actionable information, supporting decentralized healthcare systems in diagnosing complex diseases, such as LB. In this study, we developed two fluorescent polarization immunoassays (FPIAs) using IgM and IgG LB-specific synthetic epitopes/peptides to evaluate their diagnostic potential. These FPIAs showed high sensitivity and specificity in detecting IgM or IgG anti-LB antibodies in patient sera within minutes. The fluorescently labeled synthetic peptides produced significant polarization differences between infected and healthy samples, allowing clear discrimination. The FPIA-LB functions as a one-step assay, eliminating the need for secondary antibodies or complex protocols. This work highlights the FPIA technique as a robust, rapid, and efficient tool for LB diagnosis, offering a promising advance in improving early detection and patient outcomes, which could save lives and reduce long-term health complications.

Viral hemorrhagic fevers (VHFs) are severe infectious diseases caused by RNA viruses of the families Arenaviridae, Filoviridae, Flaviviridae, and Hantaviridae, characterized by high morbidity, significant case fatality rates, and frequent diagnostic uncertainty in early disease stages. For military medical services, timely clinical recognition and laboratory confirmation are essential to guide patient management, prevent nosocomial transmission, and maintain operational continuity, particularly in endemic or resource-limited deployment settings. This review critically examines current diagnostic approaches to VHF-causative agents, emphasizing their use in clinical and field medical settings. The diagnostic process, from exposure through specimen collection, laboratory testing, and result interpretation is analyzed, including the use of molecular, serological, and antigen-based assays. Particular attention is given to deployable diagnostic platforms and their role in bridging the gap between frontline clinical suspicion and definitive laboratory confirmation. Biosafety requirements and infection prevention measures are discussed as integral components of clinical diagnostic workflows, aligned with guidance from the World Health Organization and the Centers for Disease Control and Prevention. Comparative analyses of virus-specific diagnostic timelines and laboratory requirements are presented to support differential diagnosis and clinical decision-making. Emerging technologies, including rapid molecular assays and genomic methods, are evaluated for their potential to improve early diagnosis and patient outcomes. This review highlights the central role of diagnostic readiness in clinical management of the VHFs and provides evidence-based considerations to support military clinicians facing high-risk febrile illnesses in operational environments.

Angiostrongyliasis, the primary cause of eosinophilic meningitis, represents an emerging disease caused by Angiostrongylus cantonensis larvae, inadvertently transmitted to humans. The diagnosis of human angiostrongyliasis relies on epidemiological features, clinical symptoms, medical history, and laboratory findings, notably hyper eosinophilia in blood and cerebrospinal fluid. Consequently, accurate diagnosis is challenging and prone to confusion with other parasitic diseases. The quest for an early, rapid, and specific diagnostic test for angiostrongyliasis persists, driven by the imperative for enhanced test specificity. This study focused on mapping IgM epitopes on galectin-1 (Gal-1) and galectin-2 (Gal-2) proteins derived from A. cantonensis. The specificity of the epitopes was assessed using database homology analysis. After selecting specific epitopes, researchers chemically synthesized 12 individual multi-antigen peptides (MAPs4) and one chimeric polypeptide that is 65 amino acids long. The effectiveness of these synthesized peptides was subsequently evaluated using enzyme-linked immunoassay (ELISA). A total of twelve unique IgM epitopes were discovered; five were linked to Gal-1, while seven were linked to Gal-2. An ELISA-peptide method confirmed the twelve epitopes, and then the chimeric polypeptide was employed as an antigen to coat ELISA plates. This setup was evaluated with patients’ sera to diagnose strongyloidiasis in vitro. This study provides a comprehensive representation of the IgM epitopes of Gal-1 and Gal-2 from A. cantonensis. ELISA data utilizing the chimeric polypeptide demonstrate that the selected sequences hold promise for the development of a specific immunological assay tailored for the acute diagnosis of angiostrongyliasis infections.

Brucellosis is a globally prevalent zoonotic disease affecting both humans and animals. Its nonspecific clinical manifestations often complicate diagnosis, underscoring the need for reliable laboratory confirmation. Traditional serological assays, though widely used, suffer from limitations such as inconsistent sensitivity and false-positive results. To address these challenges, this study mapped IgM and IgG epitopes of the Brucella Omp-2a protein using sera from infected patients. Epitope identification was performed through SPOT synthesis on cellulose membranes, followed by assessment of potential cross-reactivity using peptide database analysis and ELISA validation. Three major IgM and seven IgG linear B-cell epitopes were identified, six of which demonstrated strong reactivity in peptide-ELISA. Importantly, no significant cross-reactivity with proteins from other human pathogens was detected. Two chimeric multi-epitope peptides, composed of 50 and 60 amino acids and integrating Brucella-specific IgM and IgG epitopes, exhibited excellent diagnostic performance in ELISA, achieving near 100% sensitivity and specificity. These findings support the potential of synthetic peptides as reliable and cost-effective alternatives to native antigens in serological assays. Further validation in larger, geographically diverse cohorts will be essential to confirm their diagnostic robustness and facilitate their integration into routine brucellosis diagnostics.

Background: Pertussis toxin (Ptx) is a major virulence factor and protective antigen of Bordetella pertussis. Understanding its antigenic landscape is essential for improving vaccine design. This study aimed to compare the linear epitope profiles of Ptx recognized by antibodies from vaccinated children and mice, identifying conserved and species-specific immune targets across subunits S1–S5. Methods: Two libraries of overlapping 14-mer peptides spanning the full-length Ptx sequence were synthesized. Sera from children and mice immunized with the whole-cell pertussis vaccine were analyzed to map antibody-binding regions. Comparative and structural analyses were performed to evaluate epitope distribution and recognition patterns. Results: Murine sera recognized 12 major epitopes, whereas children’s sera identified 24. Eleven epitopes were shared between species, mainly in subunits S1 (Ep3–5, 7, 9, 10), S3 (Ep20, 21, 25, 26), and S5 (Ep32), although minor positional shifts were observed. Eight epitopes were unique to children’s sera, located in S1 (Ep1, 6, 8), S3 (Ep22–24), and S4 (Ep27, 29–30). In the S2 subunit, four distinct epitopes were identified for each species, while only one mouse-specific epitope was detected in S4 (Ep28). Structural analysis revealed non-uniform antibody recognition, with dominant targeting of S3 and conserved antigenic hotspots, as well as selective recognition of the catalytic S1 subunit. Fourteen novel epitopes were identified. Conclusions: These findings highlight both shared and species-specific Ptx epitopes, revealing differences between murine and human immune responses. The identified conserved regions and novel epitopes provide a basis for improved pertussis vaccine design.

The onset of the COVID-19 pandemic prompted the rapid development and deployment of novel strategies and methodologies to manage the dissemination of microorganisms. Understanding the crucial role that contaminated surfaces play in the spread of viruses highlights the importance of having effective cleaning and disinfection protocols in place for inanimate objects. A variety of antimicrobial agents have shown strong effectiveness against the SARS-CoV-2 virus. Various factors can impact on the performance of these agents. As a result, technologies utilizing ozone’s microbicidal effects have been developed or improved for cleaning indoor areas, surfaces, and materials, despite ozone’s diverse uses being known for years. Ozone offers the advantage of adaptability for both gaseous and aqueous use, depending on the nature of the decontaminated surfaces. Moreover, ozone-infused water is ecologically benign, possesses microbial-fighting capabilities, and synergistically reinforces the biocidal action of other chemical disinfectants. This review aims to summarize the efforts dedicated to harnessing gaseous and aqueous ozone as a valuable means to eliminate the SARS-CoV-2 virus from environments, surfaces, clinical equipment, and office supplies. This review sourced evidence-based articles from electronic databases, including MEDLINE (via PubMed), EMBASE, the Cochrane Library (CENTRAL), and preprint repositories. The findings illustrated that ozone could serve as an additional tool for curbing the proliferation of COVID-19 and other viral infections. Additionally, we elucidated the operational attributes of ozone, the variables that influence its disinfection potency, and the mechanisms of its virucidal action. Notably, this review does not encompass the disinfection of the COVID-19 virus in wastewater.

Ozone (O₃) is a potent disinfectant, yet its efficacy in environmentally complex waters can be inconsistent. This study investigated the effectiveness of ozonation across 14 water samples (wells, rainwater, ponds) and identified key success factors. Ozone exposure (1-20 min) significantly reduced colony-forming units (CFU) in most samples, achieving complete elimination in four. However, two high-organic-load samples showed minimal change. Physicochemical analyses revealed a strong correlation between increased Oxidation-Reduction Potential (ORP) and disinfection success. We observed that the efficient generation of ozone microbubbles was the critical factor in raising the ORP to lethal levels (> ~450 mV). A paradigmatic case was sample 2, where the elimination of total coliforms (98.7%) only occurred in replicates where microbubbles formed (ORP >520 mV), failing in the replicate without microbubbles (ORP 122 mV). Mass spectrometry (MALDI-TOF) identified genera such as Pseudomonas and Bacillus, indicating the presence of microbial diversity. We conclude that ozo-nation is highly effective, but its success depends on optimizing mass transfer via microbubbles, with ORP as a fundamental real-time indicator to ensure process reliability. This study offers a practical guideline for implementing more robust and safer ozone systems, overcoming limitations observed in waters with complex contamination.

The increasing demand for advanced diagnostic technologies has positioned biosensor platforms as powerful alternatives to conventional analytical methods. Among them, lateral flow platforms (LFPs) are widely used for their speed, simplicity, and low cost. However, their limited sensitivity and lack of quantitative precision have spurred the development of enhanced systems incorporating electrochemical detection. Electrochemical biosensors offer significant advantages, including high sensitivity, excellent selectivity, and ease of miniaturization, which make them especially suitable for point-of-care testing (POCT). To address the limitations of traditional colorimetric LFPs, several strategies have been employed, such as the incorporation of nanomaterials, enzymatic amplification, and signal-enhancing labels. A particularly promising innovation is the direct integration of electrodes into LFPs, enabling real-time electrochemical readouts and enhanced analytical accuracy. Despite their potential, challenges persist, including manufacturing complexity, a lack of standardized protocols, and difficulties in scaling production for widespread adoption. Continued progress in developing hybrid platforms that combine lateral flow technology with electrochemical detection is crucial for expanding diagnostic applications in healthcare, environmental monitoring, and food safety. This work explores recent advances in electrochemical LFPs, reviewing current methodologies while discussing their advantages, limitations, and the future directions necessary to facilitate broader implementation and improve global diagnostic accessibility.