搜索结果：Toxicon : official journal of the International Society on Toxinology

Multiple System Atrophy (MSA) is a progressive neurodegenerative disorder characterized by autonomic dysfunction, parkinsonism, and cerebellar signs. Among its motor manifestations, camptocormia-defined as an abnormal forward flexion of the trunk-often emerges early in the disease course and significantly compromises patients' quality of life. Aside from physical therapy, the treatment of camptocormia remains particularly challenging, especially in MSA, where surgical approaches such as deep brain stimulation and spinal cord stimulation are generally contraindicated. This underscores the urgent need for effective, non-invasive therapeutic strategies. We report a case of MSA presenting with early-onset and severe camptocormia, who underwent sequential treatment with three non-invasive interventions: subcutaneous apomorphine injection, repetitive trans-spinal magnetic stimulation (rTMS), and botulinum neurotoxin (BoNT) injections targeting the abdominal muscles. We aimed to evaluate and compare the efficacy of each intervention using standardized clinical rating scales, instrumented gait analysis, and static posturography. All three interventions resulted in partial improvements, though with varying degrees of efficacy. Subcutaneous apomorphine injection improved walking velocity and step length but was limited by adverse effects (nausea and hypotension), which precluded the use of continuous subcutaneous infusion as a long-term therapeutic option. Spinal rTMS enhanced performance on the Timed Up and Go (TUG) test, particularly during the return phase, but had no significant effect on posture-related measures. BoNT treatment yielded the most substantial improvements in both walking velocity and step length. Moreover, it was the only intervention to improve the posture item of the Unified Multiple System Atrophy Rating Scale (UMSARS) part II. This case underscores the importance of an integrated therapeutic approach in managing camptocormia, emphasizing the role of neurophysiological and biomechanical assessments in identifying biomarkers of therapeutic response and evaluating the efficacy of treatments such as BoNT and neuromodulation.

Mycotoxins such as ochratoxin A (OTA) are hazardous chemicals produced from fungi that frequently contaminate feed ingredients, and this contamination can result in significant economic loss, disease and death. Given the persistent nature of OTA, this study was designed to provide effective and potentially scalable strategies to mitigate acute OTA toxicity using organoclay. The study explores the adsorption behavior of OTA on active binding surfaces of four organoclay composites that were synthesized by modifying sodium bentonite clay surfaces with different amounts of a hydrophobic quaternary ammonium compound (QAC). This modification serves to immobilize and stabilize the QAC. The study evaluates key thermodynamics, kinetics and binding markers including the Gibbs free energy (ΔG), adsorption capacity, enthalpy (ΔH), and binding affinity of these clays for OTA. Additionally, the protective role of these organoclays was evaluated using a sensitive ecotoxicology model. At gastric pH (pH 2), the organoclays demonstrated high adsorption capacities for OTA ranging between 0.34 and 0.51 mol kg-1 versus parent clay (0.20 mol kg-1). At intestinal pH (pH 6), the organoclay binding capacities were higher than the parent clay indicating that the modification of parent clay with QAC played a critical role in improving its adsorptive performance for OTA. Thermodynamic parameters indicated that the binding reaction was exothermic and thermodynamically favorable with ΔG values between -23.40 and -28.07 kJ mol-1 and ΔH values ranging from -13.76 kJ mol-1 to -24.31 kJ mol-1. Importantly, organoclay treatment protected hydra from OTA lethality by 95% to 100%. These findings established the proof-of-concept that OTA interactions with organoclay resulted in the prevention of OTA toxicity in hydra. Based on these findings, further studies are warranted to justify the applications and efficacy of these organoclays for short-term treatment during acute outbreaks of ochratoxicosis.

The pufferfish Takifugu rubripes accumulates the potent neurotoxin tetrodotoxin (TTX), raising significant concerns for food safety. Although TTX is known to be acquired through the food web, the molecular mechanisms underlying its accumulation remain poorly understood. In this study, we conducted a comparative proteomic analysis of the liver and skin-its primary accumulation sites-in T. rubripes following a sublethal intramuscular dose of TTX. LC/MS analysis confirmed that, after 24 h of exposure, the liver contained the highest concentration of TTX (11.80 ± 3.31 μg/g), followed by the skin (2.41 ± 0.60 μg/g). Proteomic profiling identified 361 and 282 differentially expressed proteins (DEPs) in the liver and skin, respectively. GO and KEGG pathway analyses revealed a systemic, stress-associated response. In the liver, upregulated DEPs were significantly enriched in cytoskeleton-related pathways (e.g., Focal adhesion) and inflammation/immune-related pathways (e.g., Salmonella infection), accompanied by a 1.58-fold increase in the TTX-binding protein PSTBP type 2. Downregulation of ribosome-associated proteins suggested an energy-conserving shift that reallocates resources toward defensive processes. The skin also exhibited upregulation in cytoskeleton-associated pathways (e.g., Tight junction) linked to cellular repair. However, the skin showed active upregulation of protein synthesis pathways (e.g., Aminoacyl-tRNA biosynthesis) and downregulation of the p53 signaling pathway, indicating a pro-survival, tissue-maintenance strategy. Collectively, these findings reveal tissue-specific metabolic reprogramming and highlight key candidate proteins (such as PSTBP2) that advance our understanding of TTX accumulation mechanisms and may contribute to future mitigation strategies.

The effects on human health from exposure to cyanotoxins during cyanobacterial blooms have been reported worldwide. Accumulation of toxins in fish has been documented as a potential route of transfer to humans through the ingestion of contaminated food. In this study, we determined the magnitude of microcystin (MCYSTs) concentrations in six reservoirs in central Mexico to assess the potential risk of toxin transfer to humans through the consumption of fish. Physicochemical parameters, phytoplankton composition, and MCYSTs concentrations were analyzed in water, seston, and tissue of nine fish species with varying trophic habits. We detected MCYSTs in all reservoirs, both dissolved and in seston, with the highest concentrations recorded in the Batán and Carmen reservoirs, where Microcystis was the dominant cyanobacterial genus. All fish species showed accumulation of MCYSTs, with Goodea atripinnis (phytoplanktivore) and Chirostoma jordani (zooplanktivore) exhibiting the highest concentrations in viscera and whole-body analyses. The estimated daily intake derived from consuming 100 g of contaminated fish frequently exceeded the WHO tolerable daily intake of 0.04 μg/kg body weight, indicating potential risks to human health. This finding evidences a widespread presence of hepatotoxic MCYSTs in reservoirs of central Mexico in water (regardless of the occurrence of visible blooms), seston, and accumulated in tissues of fish species. This study represents the first report of MCYSTs accumulation in fish tissue from different trophic habitats in reservoirs in central Mexico and emphasizes the need for ongoing cyanotoxin monitoring and management measures to mitigate risks to both aquatic ecosystems and human consumers.

The coconut crab, Birgus latro, is a large terrestrial decapod crustacean widely distributed throughout the tropical and subtropical islands of the Indo-West Pacific. It is consumed as food in some regions; however, cases of coconut crab poisoning have been reported. Case: On Ishigaki Island in Okinawa Prefecture, Japan, a man boiled frozen coconut crabs and consumed them for lunch and dinner. He visited our emergency department at night, on the same day, complaining of vomiting, soft feces, and dizziness. His serum digoxin level (measured using an immunoassay) was 1.8 ng/mL. Therefore, we suspected poisoning caused by coconut crab consumption and attempted to identify the causative substance using liquid chromatography-tandem mass spectrometry. Neriifolin, a cardiac glycoside, was detected in the patient's serum at a concentration of 12.8 ng/mL. Maillaud et al. previously reported an association between B. latro and Cerbera manghas, which contains neriifolin, in cases of coconut crab poisoning in New Caledonia. C. manghas is also native to Okinawa, and its distribution overlaps with areas where coconut crabs are found. As suggested by Maillaud et al., it is assumed that the coconut crab consumed in this case had accumulated neriifolin by ingesting C. manghas. Similar cases of coconut crab poisoning may occur in regions where the distributions of C. manghas and B. latro overlap. It is therefore cardinal to educate residents and tourists on the risk of poisoning from consuming coconut crabs in such regions.

Snake venom metalloproteases (SVMP) play a major role in various pathophysiological effects of snakebite envenomation, and marimastat (MMT), a selective metalloprotease inhibitor, has been investigated as a potential ancillary treatment for attenuating SVMP-mediated effects. In this work, the effect of MMT on the coagulant, hemorrhagic and neuromuscular activities of Lachesis muta (South American bushmaster) venom was assessed experimentally in vitro and in vivo and compared with therapeutic anti-Bothrops/Lachesis antivenom (AV) produced by the Instituto Butantan. MMT (0.03-3.0 mM) significantly reduced the proteolytic (caseinolytic) activity of the venom (20 μg), without affecting the esterolytic activity. The clotting activity (expressed as the minimum coagulant dose: 7.5 μg/ml) was unaffected by 0.03 mM MMT, whereas AV (antivenom:venom ratio of 1:3 v/w) completely prevented this effect. Thrombin-like activity was significantly attenuated only by AV, whereas both agents failed to prevent venom-induced factor X activation; the venom did not activate prothrombin. The venom showed α- and β-fibrinogenolytic activities. MMT partially reduced the α-chain fibrinogenolysis only at 5 min of incubation, whereas AV was ineffective; MMT abolished the β-chain fibrinogenolysis at 5 min of incubation and significantly delayed this alteration at 30 and 90 min of incubation, whereas AV produced greater inhibition of β-fibrinogenolysis after 30 and 90 min of incubation. Hemorrhagic activity was partially attenuated by MMT and abolished by AV. Venom (100 μg/ml) caused neuromuscular blockade that was attenuated by MMT alone or in combination with AV, and the resulting myonecrosis and cell vacuolization were attenuated by MMT and AV. MMT showed no additional protection when combined with AV in any of the activities tested. Together, these findings show that snake venom metalloproteases are involved to varying degrees in several activities of L. muta venom, some of which are also inhibited by therapeutic antivenom.

Brown spider envenomation (loxoscelism) represents a significant public health concern in South America, yet most studies focus on a few medically recognized species. Here, we provide the first molecular and functional characterization of the venom gland extract from Loxosceles aff. variegata (LafvVGE), a brown spider collected in synanthropic habitats in Ituiutaba, Minas Gerais, Brazil. SDS-PAGE and immunoblot analyses revealed prominent protein bands consistent with phospholipase D (PLD) toxins, the main agents of loxoscelism symptoms. ELISA assays demonstrated that LafvVGE is effectively recognized by Brazilian therapeutic antivenoms, indicating immunological cross-reactivity. Enzymatic assays confirmed sphingomyelinase and collagenase/gelatinase activities comparable to those of Loxosceles gaucho (a species of acknowledged medical relevance), although LafvVGE from female individuals showed higher activities than male derived pools under our experimental conditions. Neutralization assays showed complete inhibition of sphingomyelinase activity but only partial inhibition of gelatinase activity by the anti-loxoscelic antivenom, highlighting differential susceptibility of venom components to antivenom-mediated neutralization under in vitro conditions. Molecular analysis of venom gland transcripts identified eight distinct PLD isoforms (LafvPLD1-8), all containing conserved catalytic and metal-binding residues characteristic of class II Loxosceles PLDs. Structural modeling revealed isoform-specific variations in the aromatic cage motif and electrostatic surface, suggesting potential effects on membrane interactions and substrate specificity. Collectively, these findings place L. aff. variegata within the biochemical and structural spectrum of medically relevant Loxosceles species, expanding comparative knowledge of PLD diversity and function. While clinical relevance remains to be established in vivo, this study underscores the value of integrating biochemical, immunological, and structural analyses to identify emerging venom phenotypes with potential implications for surveillance and antivenom coverage.

The use of alternative methods for in vivo lethality neutralization assays in mice (potency tests) during antivenom production has been encouraged by the World Health Organization, based on the 3Rs principle (Replacement, Reduction, and Refinement). In addition to animal welfare-associated benefits, alternative methods can reduce the time, cost, and intrinsic variability linked with animal-based assays. Several studies have been conducted using different methodologies, including immunological assays and tests for inhibiting enzymatic and toxic activities of venom, to assess their level of agreement with the in vivo method, which is considered the gold standard. However, the results varied depending on the methodology applied, as well as the specificity and avidity of antibodies toward the toxic and non-toxic venom fractions, and the composition of each venom. This literature review examined these studies and their outcomes, revealing that the Enzyme-Linked Immunosorbent Assay (ELISA) has historically been the most evaluated alternative to the in vivo antivenom potency test. Despite promising results, the development and implementation of substitute methodologies face significant challenges, primarily in their validation, which requires substantial time and resources from the institutions that produce them. Efforts should focus on developing, validating, and applying alternative methods for potency assays in antivenom production, particularly in the early and intermediate stages, where regulatory requirements are less stringent.

The recent clinical development of botulinum neurotoxin serotype E (BoNT/E), valued for its rapid onset, introduces a novel therapeutic protein with an unknown long-term immunogenic risk profile. To generate a method-agnostic risk assessment, we applied a consensus computational immunogenicity framework, rigorously calibrated against the clinically observed 1-3% neutralizing antibody (NAb) incidence for BoNT/A. Our multi-platform strategy integrated an ensemble of four independent HLA class II epitope prediction algorithms, triplicate molecular dynamics simulations using distinct force fields, and three independent systems immunology models. This triangulated approach consistently identified BoNT/E as possessing a significantly enriched epitope landscape, with 73-83% more predicted strong HLA binders than BoNT/A. Biophysical simulations confirmed that BoNT/E-derived peptides form more stable complexes with HLA molecules, exhibiting a mean ΔΔG ∼ binding ∼ advantage of approximately -13 kcal/mol. Systems-level models projected a consensus threefold increase in the hazard for NAb development (HR = 3.03) and an accelerated risk for concomitant BoNT/A + E therapy. Control analyses confirmed the specificity of the signal to native BoNT/E epitope architecture, and Bayesian modelling quantified a >99% posterior probability that BoNT/E confers higher relative immunogenic risk. These predictions remain subject to the inherent simplifications of computational models relative to the complexity of human immune responses. Nonetheless, this convergent, cross-platform evidence establishes a robust risk hypothesis, underscoring the need for enhanced clinical immunogenicity monitoring for BoNT/E.

The alkaloids swainsonine and calystegines are found in some Ipomoea species. Swainsonine is a potent cellular glycosidase inhibitor that often poisons livestock while the calystegines inhibit cellular glycosides and can cause cellular glycoside dysfunction. The role of the calystegines in a livestock model is lacking as no studies to date have experimentally dosed calystegines alone in livestock. The objective of this study was to directly compare goats dosed with Ipomoea carnea containing both swainsonine and calystegines and I. carnea containing only the calystegines. Three groups of four goats were treated with ground alfalfa (control), I. carnea containing both swainsonine and calystegines or I. carnea containing only the calystegines, for 45 days. Animals were observed daily; body weights, serum biochemistries, and serum swainsonine concentrations were determined weekly. On day 45, all animals were euthanized and necropsied. Goats treated with I. carnea containing only the calystegines showed no changes in clinical disease, serum biochemistries, and histologic lesions while goats treated with swainsonine and the calystegines developed clinical disease and showed changes in serum biochemistries and histologic lesions characteristic of animals poisoned by other plants containing swainsonine. The data suggests that the calystegines play no role in field cases or experimental studies for plants containing swainsonine and the calystegines. Swainsonine is the principal driver of toxicity in plants containing both these toxins.

Oxyrhopus guibei is a conspicuous colubrid snake popular in the exotic pet trade that has been implicated in envenomation cases involving local and systemic clinical manifestations. This study aims to characterize the venom apparatus of O. guibei, including the composition and immunochemical reactivity of its secretion. The maxilla and fangs were examined using scanning electron microscopy, while the histology and histochemistry of the glands were analyzed via hematoxylin-eosin and periodic acid-Schiff (PAS), Alcian Blue pH 2.5, and Gomori's trichrome staining. Additionally, the protein profile was evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Tricine-SDS-PAGE, and cross-reactivity with bivalent and tetravalent bothropic antivenoms was assessed by Western blot. Anatomically, recurved teeth were observed in the anterior maxilla, followed by enlarged, deeply grooved posterior teeth (opisthoglyphous dentition). Histologically, Duvernoy's venom glands exhibited distinct structural and chemical characteristics compared to the supralabial glands; their secretory units are organized into lobules of predominantly serous acini composed of columnar cells with acidophilic cytoplasm and PAS-negative basal nuclei. The venom protein profile displayed bands ranging from ≈8.5 to 65 kDa, with a predominance of proteins migrating within molecular mass ranges consistent with snake venom metalloproteinases, which showed immunoreactivity with both antivenoms tested. In conclusion, O. guibei possesses a specialized venom delivery system consisting of modified anatomical structures designed to deliver a venom containing components reminiscent of bothropic venoms, primarily involved in prey subjugation and with the potential to cause envenomation in humans. This study provides preliminary insights into the venom of this species and underscores the need for further functional characterization to determine its potential toxicological relevance to humans.

Legal use of herbal health products must comply with quality and safety standards required by health regulatory authorities in order to protect consumers from risks associated with misidentification, falsification and undesirable toxic effects. Some plant-derived compounds can act as CYP450 inhibitors, potentially leading to herb-drug interactions and, in some cases, direct hepatotoxicity, particularly when CYP450-mediated bioactivation is involved. There is a critical need to develop reliable and efficient screening assays to identify potential CYP450 inhibitors in such complex mixtures and hepatotoxicity risks associated to CYP450 metabolization. In this context, we propose associating a HepaRG cell-based test with suitable qualitative and quantitative phytochemical analyses. A case study was conducted using Tinospora crispa (T. crispa) stems harvested in Laos ("boraphet"). This herb is traditionally used for its antidiabetic effects and is known to cause hepatotoxicity, most likely due to the presence of furanoditerpenoids bioactivated by cytochromes. A Molecular Networking-based UHPLC-(HRMS)2 fingerprint of furanoditerpenoids was established by analysing methanolic extracts, enriched fractions of a representative sample and standards, by applying the bioinformatic Feature-Based Molecular Networking workflow. The fingerprinting model enabled identification and comparison between three samples harvested from nearby geographical locations (Hinboun, Yommalath and Nakaiy) through the detection of fourteen furanoditerpenoids. Four of them were identified as borapetosides A-D, eight had not yet been isolated, and were tentatively assigned. Borapetoside C content, determined by UHPLC-UV is proposed as an assay for the quality control of T. crispa stems. The CYP3A4 inhibitory activities of the methanol extracts and of borapetoside C were measured in a HepaRG cell model, which is a validated model for studying CYP450 inhibitory effects. All extracts and borapetoside C exhibited CYP3A4 inhibition. This work highlights the interest in including a HepaRG cell-based test in the T. crispa stem specification to assess the toxicity of herbal constituents associated with CYP450 inhibition.

Traditional practices such as the application of black stone (BS) following snakebite are common in rural settings of Asia, Africa and Latin America. However, the use of black stone remains controversial. While proponents of BS claim that it absorbs venom to mitigate systemic envenoming, critics raise concerns regarding its safety and efficacy. The World Health Organisation guidelines advise against its use, yet reliance on this practice persists. This narrative review aims to synthesise and provide comprehensive evidence on BS, emphasising its identity, proposed mechanism, efficacy, associated risks, and the potential drivers of its continued use. The review involved peer-reviewed biomedical literature retrieved from PubMed and Google Scholar using appropriate search terms. It focused on experimental studies, reviews, and observational studies. The precise composition of BS remains unclear. However, studies suggest that it is derived from burned animal horn, bone, or stone. Its purported mechanism involves venom absorption at the bite site, although this remains unproven. Animal studies and WHO guidelines do not support its clinical use in the management of snakebite. Its persistent use is potentially driven by socioeconomic, cultural, and health system factors. In conclusion, BS currently lacks validated therapeutic benefits and may contribute to delays in seeking appropriate hospital treatment, as well as cause infections and tissue injuries. Snakebite-affected areas should improve community and school education to nullify dependence on traditional post-snakebite practices, including the use of black stone, and engage with traditional healers in a culturally sensitive manner to ensure timely access to antivenom.

Scorpion stings remain a significant public health problem in tropical regions, particularly in Brazil, where an estimatied 117,185 cases are reported annually, reflecting a persistent and growing burden on the healthcare system. Globally, more than two million cases occur each year, especially in areas undergoing environmental disturbances such as deforestation and unplanned urbanization. This study investigates the ecological and social contexts of scorpion stings in Manaus, the largest metropolis of the Brazilian Amazon. This qualitative study was conducted at the Dr. Heitor Vieira Dourado Tropical Medicine Foundation (FMT-HVD), a reference center for scorpion stings, between January 2020 and March 2025. Data were collected through clinical questionnaires, in-depth interviews, and household field observations. Thematic analysis was performed using Atlas.ti. Thirty participants from different age groups were included. Most cases were clinically mild. Five themes emerged from the qualitative analysis that address perceptions and reactions to the sting, environmental risk factors, post-accident behaviors, emotional impacts, and patient care pathways. Participants reported overlap between domestic spaces and forest fragments, limited knowledge about prevention, emotional distress, and barriers to accessing care, including late referrals and transportation difficulties. Scorpion stings in the Amazon are shaped by environmental, social, and structural factors. Integrated strategies involving environmental management, health education, urban planning, and improved healthcare access are essential to reduce their burden.

Actinoporins are a protein family of α-pore-forming proteins (PFPs), initially isolated from sea anemones. These proteins are key components of the venom system used by sea anemones and some other cnidarians, the most ancient extant lineage of venomous animals, where they play a role in predation and predator deterrence through rapid membrane disruption. The scientific journey to understand these potent molecules began more than five decades ago with their initial isolation from crude anemone extracts. In recent years structural studies enabled the determination of multiple structures of monomeric actinoporins in solution and in complexes with ligands, elucidating their mechanism of membrane binding and pore formation, highlighting in the fragaceatoxin C (FraC) pore structure. Recently, two groups independently produced multiple pore structures of actinoporins and a close homologue, further elucidating the mechanism of pore formation and investigating the role of lipids in the pore structure, which is even more prominent than initially thought. Hereby, we highlight the achievements of the two studies and discuss some remaining open questions.

From the monotypic family Superstitioniidae, the scorpion Superstitionia donensis presents a high diversity of transcripts encoding non-disulfide-bridged peptides (NDBPs). In this study, five peptide sequences inferred from a previous venom gland transcriptomic analysis were selected based on sequence similarity and physicochemical properties associated with previously antimicrobial peptides. An additional peptide was designed to evaluate the effect of changes on selected physicochemical properties on its hemolytic and antimicrobial activity. The selected peptides were synthesized using Merrifield Solid-Phase peptide synthesis and evaluated form antimicrobial activity against Gram-negative and Gram-positive bacteria, belonging to the ESKAPE group and yeasts of the Candida and Cryptococcus genera, as well as for hemolytic and cytotoxic activity in mammalian cells. Some peptides showed antimicrobial activity against selected microbial strains, including Escherichia coli, Enterococcus faecium, Staphylococcus aureus, Candida krusei, and Candida tropicalis, while limited or no activity was observed against Klebsiella pneumoniae, Pseudomonas aeruginosa, and Candida glabrata. Notably, treatment with some selected peptides improved the survival of Galleria mellonella larvae infected with S. aureus, supporting their functionality in vivo. Peptide toxicity was also evaluated in human erythrocytes and two human cell lines, showing that some peptides with antimicrobial activity also exhibited hemolytic effects; however, they showed low cytotoxicity to cell lines at antimicrobial concentrations. Despite observed hemolytic activity, the relatively low cytotoxicity against human cell lines suggests potential for improvement through structural refinement. These results demonstrate that S. donensis NDBPs exhibit antimicrobial activity and hemolytic effect, and their sequences may serve as templates for further modification. Furthermore, the findings reveal that antimicrobial potency and cytotoxicity are influenced by sequence similarity, net charge, and hydrophobicity. Nevertheless, the physicochemical property values required to identify an optimal antimicrobial peptide remain poorly defined, as several parameters exhibit overlap between high- and low-performing candidates, which highlights a key challenge for their therapeutic development.

This study presents a structured process design for a polyvalent veterinary clostridial vaccine along with a comprehensive techno-economic analysis. Four scenarios were evaluated by examining the primary factors: (i) cultivation equipment (stainless-steel (SS) versus single-use bioreactor (SUB)), (ii) options for the inactivation and formulation processes (1500-L and 250-L SS vessels, or 750-L SUB bags), (iii) in process quality control strategies (conventional 14-day sterility testing versus rapid 1-day testing), and (iv) a closed-system design for formulation and filling. The base scenario (characterized by high demand for 250-L SS equipment, long sterility assays, and an open system design) indicated a production capacity of 32 million doses with unfavorable economics (ROI -65%, gross margin -47%, and NPV -5.0 million USD over 15-year lifetime). Introduction of rapid sterility testing across three alternative scenarios yielded identical process durations and an annual output of 51.2 million doses, but with distinct capital and operating cost profiles. Notably, the use of single-use components reduced capital costs and water consumption, but totally resulted in negative economic indicators. Another scenario (scenario 3), featuring fewer SS equipment and a closed-system design, yielded a positive NPV (∼3.66 million USD), a payback of 1.4 years, a gross margin of 35%, and ROI of 23%. Sensitivity analysis revealed that NPV is most influenced by the product selling price, followed by labor, material, and utility costs. With a ±25% variation in various factors, only the optimized scenario (scenario 3) maintained profitability. Collectively, scenario 3 emerged as the most viable option for sustained industrial clostridial vaccine production.

Botulinum neurotoxin type A (BoNT/A) is a widely used biologic therapeutic, yet the influence of natural toxin sequence variation on the stability of neutralizing antibody binding remains poorly understood. In this study, we performed a comprehensive computational analysis to quantitatively assess antibody binding robustness across naturally occurring BoNT/A variants. Using a curated dataset spanning all major subtypes, we integrated sequence variability mapping, ensemble antibody docking, explicit-solvent molecular dynamics simulations (totalling >30 &#x3bc;s), Bayesian hierarchical modelling, and interface network analysis. Sequence analysis revealed that amino acid variability is strongly enriched in solvent-exposed regions, with mean Shannon entropy increasing from 0.07 in the light-chain core to 0.48 in surface loops of the heavy chain, and a significant association between entropy and solvent accessibility (Wilcoxon p&#x202f;<&#x202f;10-6). Docking showed modest variant-dependent energetic dispersion (Cohen's d&#x202f;<&#x202f;0.5) that poorly correlated with dynamic stability metrics (&#x3c1;&#x202f;=&#x202f;0.18-0.31). In contrast, molecular dynamics simulations identified large variant-specific effects on interface stability, with multiple variants exhibiting >1 standard deviation increases in interface root-mean-square deviation (RMSD, a measure of average atomic displacement between structures) and reductions in contact persistence and hydrogen-bond occupancy. A composite Binding Robustness Index (BRI) separated high- and low-robustness variants by 2.1 to 2.7 standard deviations within antibody systems. Bayesian modelling indicated that 31-46% of variants had high posterior probability (>0.95) for reduced binding robustness. Interface network analysis revealed that low-robustness variants consistently disrupted cooperative interface topology, with 35-61% loss of high-betweenness residues and increased modularity, strongly correlating with Bayesian effect estimates (&#x3c1;&#x202f;=&#x202f;0.74). These results demonstrate that natural BoNT/A sequence variation leads to substantial, antibody-specific differences in binding robustness, driven by localized disruption of cooperative interface networks rather than uniform binding weakening. This study provides a quantitative computational framework for assessing antibody binding stability across toxin variants, with implications for therapeutic durability and resistance mechanisms.

Centruroides mascota scorpion was described in 2011 by Ponce Saavedra's group as a species of medical importance in the state of Jalisco. In this study, we characterized two populations of scorpions collected in the states of Jalisco and Nayarit. Through a morphometric analysis, the determination of the LD50 of the respective venoms, electrophoresis, separation of their components by HPLC, mass spectrometry, and neutralization tests, it has been possible to establish that they correspond to the same species, distributed in different habitats. The scFv 10FG2 antibody fragment, which shows broad cross-neutralization of various scorpion toxins, was evaluated for the first time in the recognition of toxins from this venom and in its neutralization. The recombinant antibody fragment 10FG2 neutralizes 2 LD50 of venom of Centruroides mascota from both localities.