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Adjuvants enhance, prolong, and modulate immune responses by vaccine antigens to maximize protective immunity and enable more effective immunization in the young and elderly. Most adjuvants are formulated with injectable vaccines. However, an intranasal route of vaccination may induce mucosal and systemic immune responses for enhancing protective immunity in individuals and be easier to administer compared to injectable vaccines. In this study, a next generation of broadly-reactive influenza hemagglutinin (HA) vaccines were developed using the Computationally Optimized Broadly Reactive Antigen (COBRA) methodology. These HA vaccines were formulated with Mastoparan 7 (M7-NH2) mast cell degranulating peptide adjuvant and administered intranasally to determine vaccine-induced seroconversion of antibodies against a panel of influenza viruses and protection following infection with H1N1 and H3N2 viruses in mice. Mice vaccinated intranasally with M7-NH2-adjuvanted COBRA HA vaccines had high HAIs against a panel of H1N1 and H3N2 influenza viruses and were protected against both morbidity and mortality, with reduced viral lung titers, following challenge with an H1N1 influenza virus. Additionally, M7-NH2 adjuvanted COBRA HA vaccines induced Th2 skewed immune responses with robust IgG and isotype antibodies in the serum and mucosal lung lavages. Overall, this intranasally delivered M7-NH2 -adjuvanted COBRA HA vaccine provides effective protection against drifted H1N1 and H3N2 viruses. [ABSTRACT FROM AUTHOR]

Northern Fennoscandia entered Christian Dotremont's (1922–1979) imagination in 1956. The Belgian avant‐gardist was comfortable in Central‐European artistic milieus through his involvement in CoBrA (1948–1951), but a total of 12 journeys to Sápmi between 1956 and 1978 had a profound effect on his creative work, especially the logograms he is best known for. This article studies Dotremont's travel writings and logograms as site‐specific forms of writing, which can be seen as precursors to ecopoetic approaches. Dotremont's journeys took place at a cultural turning point, when ethnographers had made their field trips but mass tourism was still in its embryonic state in Sápmi. His Sápmi‐inspired travel writings reveal how the idealism related to a hyperborean north initially intrigued him while he sought to elude modernity. Dotremont's cultural exchanges in Sápmi were non‐artistic but manifested in his art and writing. Sápmi brought about an ecological awakening through awe that was not sublimated but a lived experience. Dotremont immersed himself in Sápmi, with fundamental repercussions to his creative exploits. [ABSTRACT FROM AUTHOR]

Snake venom L-amino acid oxidases (LAAOs) are flavoenzymes with diverse physiological and pharmacological effects. These enzymes are found to showcase anticoagulant, antiplatelet, cytotoxicity and other biological effects in bite victims. However, the exact mechanism through which they exhibit several biological properties is not yet fully understood. The current study focussed on the purification of cobra venom LAAO and the functional characterization of purified LAAO. A novel L-amino acid oxidase NNLAAO70 with a molecular weight ~70 kDa was purified from the venom of an Indian spectacled cobra (Naja naja). NNLAAO70 showed high substrate specificity for L-His, L-Leu, and L-Arg during its LAAO activity. It inhibited adenosine di-phosphate (ADP) and collagen-induced platelet aggregation process in a dose-dependent manner. About 60% inhibition of collagen-induced and 40% inhibition of ADP-induced platelet aggregation was observed with a 40 μg/ml dose of NNLAAO70. NNLAAO70 exhibited bactericidal activity on Bacillus subtilis, Escherichia coli, Bacillus megaterium, and Pseudomonas fluorescens. NNLAAO70 also showed cytotoxicity on A549 cells in vitro. It showed severe bactericidal activity on P. fluorescens and lysed 55% of cells. NNLAAO70 also exhibited drastic cytotoxicity on A549 cells. At 1 μg/ml dosage, it demonstrated a 60% reduction in A549 viability and induced apoptosis upon 24-h incubation. H2O2 released during oxidative deamination reactions played a major role in NNLAAO70-induced cytotoxicity. NNLAAO70 significantly increased intracellular reactive oxygen species (ROS) levels in A549 cells by six fold when compared to untreated cells. Oxidative stress-mediated cell injury is the primary cause of NNLAAO70-induced apoptosis in A549 cells and prolonged oxidative stress caused DNA fragmentation and activated cellular secondary necrosis. [ABSTRACT FROM AUTHOR]

Background: Snakebite envenomation inflicts a high burden of mortality and morbidity in sub-Saharan Africa. Antivenoms are the mainstay in the therapy of envenomation, and there is an urgent need to develop antivenoms of broad neutralizing efficacy for this region. The venoms used as immunogens to manufacture snake antivenoms are normally selected considering their medical importance and availability. Additionally, their ability to induce antibody responses with high neutralizing capability should be considered, an issue that involves the immunization scheme and the animal species being immunized. Methodology/Principal findings: Using the lethality neutralization assay in mice, we compared the intrageneric neutralization scope of antisera generated by immunization of horses with monospecific, bispecific/monogeneric, and polyspecific/monogeneric immunogens formulated with venoms of Bitis spp., Echis spp., Dendroaspis spp., spitting Naja spp. or non-spitting Naja spp. It was found that the antisera raised by all the immunogens were able to neutralize the homologous venoms and, with a single exception, the heterologous congeneric venoms (considering spitting and non-spitting Naja separately). In general, the polyspecific antisera of Bitis spp, Echis spp, and Dendroaspis spp gave the best neutralization profile against venoms of these genera. For spitting Naja venoms, there were no significant differences in the neutralizing ability between monospecific, bispecific and polyspecific antisera. A similar result was obtained in the case of non-spitting Naja venoms, except that polyspecific antiserum was more effective against the venoms of N. melanoleuca and N. nivea as compared to the monospecific antiserum. Conclusions/Significance: The use of polyspecific immunogens is the best alternative to produce monogeneric antivenoms with wide neutralizing coverage against venoms of sub-Saharan African snakes of the Bitis, Echis, Naja (non-spitting) and Dendroaspis genera. On the other hand, a monospecific immunogen composed of venom of Naja nigricollis is suitable to produce a monogeneric antivenom with wide neutralizing coverage against venoms of spitting Naja spp. These findings can be used in the design of antivenoms of wide neutralizing scope for sub-Saharan Africa. Author summary: Parenteral administration of antivenoms is the core of the current treatment of snakebite envenomations, and there is an urgent need to produce antivenoms of wide neutralizing efficacy for sub-Saharan Africa. The active substance of antivenoms are antibodies (or antibody fragments) purified from plasma of horses or sheep immunized by the repeated injection of snake venoms. Generally, these antibodies can neutralize the venoms used as immunogens and other related venoms. Normally, the venoms used as immunogens are selected considering their medical importance and availability. To complement these criteria with information regarding the immunogenicity of venoms, we compared monospecific, bispecific/monogeneric, and polyspecific/monogeneric antisera towards venoms of Bitis spp., Echis spp., Dendroaspis spp., spitting Naja spp. or non-spitting Naja spp, regarding their intrageneric neutralization scope, evaluated by the lethality neutralization assay in mice. We found that the polyspecific antisera against venoms of Bitis spp, Echis spp, Dendroaspis spp, or non-spitting Naja gave the best neutralization profile. On the other hand, the monospecific, bispecific and polyspecific antisera towards venoms of spitting Naja venoms showed a similar performance. This information suggests that polyspecific immunogens could be the best alternative to produce antivenoms with the widest neutralizing coverage against sub-Saharan African snake venoms. [ABSTRACT FROM AUTHOR]

The current business climate has generated considerable uncertainty and disrupted supply chain processes. Suppliers have frequently been identified as the primary source of hazards responsible for supply chain disruptions. Using a strategic approach to supplier selection that prioritizes providers with resilience features, mitigating the risk exposure inherent in supply chains is possible. This study proposes a comprehensive gray multiple-criteria decision making (MCDM) method incorporating resilience attributes to supplier selection. To determine criteria weights, the gray PSI and gray BWM methodologies were used, and to evaluate and prioritize resilient providers, the gray MCRAT and gray COBRA methodologies were applied. According to the results obtained by the suggested methodology, the supplier that demonstrated the greatest degree of resilience was determined to be the provider categorized as SPIR 4. The sequential sequence of the SPIR numbers is as follows: SPIR 5, SPIR 1, SPIR 3, SPIR 2, and SPIR 6. The data demonstrate that the developed approach produced accurate results. [ABSTRACT FROM AUTHOR]

Snakebite envenoming is a neglected tropical disease that causes substantial mortality and morbidity globally. The venom of African spitting cobras often causes permanent injury via tissue-destructive dermonecrosis at the bite site, which is ineffectively treated by current antivenoms. To address this therapeutic gap, we identified the etiological venom toxins in Naja nigricollis venom responsible for causing local dermonecrosis. While cytotoxic three-finger toxins were primarily responsible for causing spitting cobra cytotoxicity in cultured keratinocytes, their potentiation by phospholipases A2 toxins was essential to cause dermonecrosis in vivo. This evidence of probable toxin synergism suggests that a single toxin-family inhibiting drug could prevent local envenoming. We show that local injection with the repurposed phospholipase A2-inhibiting drug varespladib significantly prevents local tissue damage caused by several spitting cobra venoms in murine models of envenoming. Our findings therefore provide a therapeutic strategy that may effectively prevent life-changing morbidity caused by snakebite in rural Africa. [ABSTRACT FROM AUTHOR]

The posterior salivary gland of Octopus vulgaris is apocrine and has a discrete character in the animal kingdom. In general, studying the evolution of any toxic organism typically starts with an examination of its defense system, including the minute structures with similar organisms across various phyla in the animal kingdom. The posterior salivary gland of Octopus vulgaris consists of a thick gland wall with a contraction function for pushing the toxin through the gland duct and into the pass tube, which opens in a buccal mass. The posterior salivary gland of Octopus vulgaris secretes toxic saliva, which is thermostable liquid resistant to heat and has a darkened blue appearance when stained with bromophenol blue, more pronounced than the venom. The present gland consists of nine types of cells associated with a canal that transports the toxin from the posterior to anterior salivary glands. These glands are located in the middle of the gland and consist of a mixture of circular and longitudinal muscle fibers, respectively. The secretion of the present gland is considered analogous to the evolutionary progression from octopus to the cobra snake. In the cobra snake, there are two types of cells with thick walls that aid in the contraction of the gland to push the toxin outside the mouth. In octopuses, two small cells represent an early evolutionary stage observed in glands across the animal kingdom, as recorded in the earliest records. [ABSTRACT FROM AUTHOR]

Animal colour is a highly adaptive phenotypic trait that can respond to several selection pressures, including those facilitated by variations in climate. The thermal melanism hypothesis predicts that for ectotherms, selection for darker phenotypes should be highest in cooler areas because darker pigmentation should provide thermoregulatory advantages. We tested whether intraspecific variation in colour distribution of Cape Cobras (Naja nivea) conforms to this hypothesis using a dataset of ∼800 georeferenced photographs of cobras available from community science platforms. We scored the dorsal colouration of snakes in each photograph and tested for associations between snake colours and climate variables at those locations. Our results provide strong evidence to suggest that temperature and solar radiation are important predictors of N. nivea colour occurrence, whereas elevation and precipitation are not. Overall, darker snakes have a significantly higher probability of occurrence in colder areas with low solar radiation than lighter snakes, which are more likely to occur in hotter areas with more solar radiation. Our study is the first to provide evidence for thermal melanism in a sub-Saharan African snake species, which was made possible by the availability of community science data. [ABSTRACT FROM AUTHOR]

The evolutionary interplay between predator and prey has significantly shaped the development of snake venom, a critical adaptation for subduing prey. This arms race has spurred the diversification of the components of venom and the corresponding emergence of resistance mechanisms in the prey and predators of venomous snakes. Our study investigates the molecular basis of venom resistance in pythons, focusing on electrostatic charge repulsion as a defense against α-neurotoxins binding to the alpha-1 subunit of the postsynaptic nicotinic acetylcholine receptor. Through phylogenetic and bioactivity analyses of orthosteric site sequences from various python species, we explore the prevalence and evolution of amino acid substitutions that confer resistance by electrostatic repulsion, which initially evolved in response to predatory pressure by Naja (cobra) species (which occurs across Africa and Asia). The small African species Python regius retains the two resistance-conferring lysines (positions 189 and 191) of the ancestral Python genus, conferring resistance to sympatric Naja venoms. This differed from the giant African species Python sebae, which has secondarily lost one of these lysines, potentially due to its rapid growth out of the prey size range of sympatric Naja species. In contrast, the two Asian species Python brongersmai (small) and Python bivittatus (giant) share an identical orthosteric site, which exhibits the highest degree of resistance, attributed to three lysine residues in the orthosteric sites. One of these lysines (at orthosteric position 195) evolved in the last common ancestor of these two species, which may reflect an adaptive response to increased predation pressures from the sympatric α-neurotoxic snake-eating genus Ophiophagus (King Cobras) in Asia. All these terrestrial Python species, however, were less neurotoxin-susceptible than pythons in other genera which have evolved under different predatory pressure as: the Asian species Malayopython reticulatus which is arboreal as neonates and juveniles before rapidly reaching sizes as terrestrial adults too large for sympatric Ophiophagus species to consider as prey; and the terrestrial Australian species Aspidites melanocephalus which occupies a niche, devoid of selection pressure from α-neurotoxic predatory snakes. Our findings underline the importance of positive selection in the evolution of venom resistance and suggest a complex evolutionary history involving both conserved traits and secondary evolution. This study enhances our understanding of the molecular adaptations that enable pythons to survive in environments laden with venomous threats and offers insights into the ongoing co-evolution between venomous snakes and their prey. [ABSTRACT FROM AUTHOR]

Delhi, the capital city of India is, highly urbanized and surrounded by remnant forest, farms, ridges, and other green areas experience regular snake encounters in and around residential, institutional, and industrial areas. A total of 41 months of sampling from January 2019 to May 2022 was conducted wherein we, studied the snake assemblage in Delhi to determine the species composition, encounter frequency, seasonal activity patterns, and probable encounter sites in an urban setup. We documented 372 individuals belonging to 15 species from seven families out of 23 species found in Delhi. Snakes were found inside forests, public parks, homes, drain networks, streets, office buildings, and even in school-college buildings. The most recorded species being Ptyas mucosa (37.37%, n = 139), Naja naja (19.62%, n = 73), and Lycodon aulicus (13.44%, n = 50). The highest numbers of incidents were reported in the month of July (22.04%, n = 82) and August (19.89%, n = 74) during the peak monsoon season, for identifying high encounter sites, we used a geostatistical modeling tool, Ordinary kriging to identify places having more snake occurrences. We further used a statistical spatial method called average nearest neighbor distance to detect the pattern distribution of snake species. Spatial interpolation done through Ordinary kriging highlighted two areas having concentrated snake encounters. The results of the average nearest neighbor distance analysis showed three species having clustered and two species having dispersed distribution. The incidence of snake encounters was found to be highly seasonal and appeared to be associated mainly with monthly rainfall, temperature, and humidity. The findings of this study on snakes' distribution patterns provide valuable insights into the conservation of these species. Understanding their habitat preferences and spatial distribution is crucial for the implementation of effective conservation strategies. [ABSTRACT FROM AUTHOR]

Background: Snakebite is an important occupational hazard in India as it is always been a land of poisonous snakes. The isssue which the physician confronts while treating a snake bite patient is assesment of degree of envenomation and requirement of ASV dose. Aims and Objectives: This study was taken to evaluate the optimum dose of anti-snake venom (ASV) required based on the severity of envenomation. Materials and Methods: Patients with a history of snakebite brought to the Department of General Medicine, Hassan, were included in this study. The study was conducted during the period from December 2014 to June 2016. The sample size of 80 patients was included in the study after fulfilling the inclusion criteria. Results: A total of 80 patients were included in the study. The majority of the victims were males (70%), age between 21 and 40 years (47.5%), and agriculture was the main occupation (67.3%). 51.25% did not identify the snake. The most poisonous were viper and cobra types which were 36.5% and 12.5%, respectively. A delay in lag time of 8.99±8.2 h was observed in severe envenomation. Overall, 51 cases (63.75%) had cellulitis, and 3 (3.75%) in the severe group required fasciotomy. Twenty percent had hematologic derangements. Ten percent of patients had developed renal failure and one required dialysis. 8.75% of patients developed respiratory failure and all required mechanical ventilation. The average dose of ASV vials used in mild, moderate, and severe envenomation was 9.04±3.51 vials, 18.5±5.27 vials, and 28.6±7.30 vials, respectively. The overall mortality rate was 5%. Conclusions: The optimum dose of ASV required in mild, moderate, and severe envenomation is 9.04±3.51 vials, 18.5±5.27 vials, and 28.6±7.30 vials, respectively to neutralize the circulating venom and lower the risk of development of serious complications. [ABSTRACT FROM AUTHOR]

Snakes contain three types of phospholipase A2 (PLA2)-inhibitory proteins in their blood, PLIα, β, and γ, which protect them from their own venom, PLA2. PLIβ is the snake ortholog of leucine-rich α2 glycoprotein (LRG). Since autologous cytochrome c (Cyt c) serves as an endogenous ligand for LRG, in this study, we purified snake LRGs from various snake serum samples using Cyt c affinity chromatography. All purified snake LRGs were found to be dimers linked by disulfide bonds. Laticauda semifasciata and Naja kaouthia LRGs showed no inhibitory activity against L. semifasciata PLA2 and weak inhibitory activity against Gloydius brevicauda basic PLA2. Elaphe climacophora PLIβ had weaker inhibitory activity against G. brevicauda basic PLA2 than G. brevicauda and Elaphe quadrivirgata PLIs, which are abundant in blood and known to neutralize G. brevicauda basic PLA2. Protobothrops flavoviridis LRG showed no inhibitory activity against basic venom PLA2, PL-X, or G. brevicauda basic PLA2. Binding analysis of P. flavoviridis LRG using surface plasmon resonance showed very strong binding to snake Cyt c, followed by that to horse Cyt c, weak binding to yeast Cyt c, and no binding to P. flavoviridis PL-X or BPI/II. We also deduced the amino acid sequences of L. semifasciata and P. flavoviridis LRG by means of cDNA sequencing and compared them with those of other known sequences of PLIs and LRGs. This study concluded that snake LRG can potentially inhibit basic PLA2, but, whether it actually functions as a PLA2-inhibitory protein, PLIβ, depends on the snake. [ABSTRACT FROM AUTHOR]

Toll-like receptor 4 (TLR4) is a crucial inflammatory signaling pathway that can serve as a potential treatment target for various disorders. A number of inhibitors have been developed for the TLR4 pathway, and although no inhibitors have been approved for clinical use, most have been screened against the TLR4-MD2 conformation. The venom gland is the organ of venomous snakes that secretes substances that are toxic to other animals. The level of gene transcription in venom glands is different from that in other tissues, includes a large number of biologically active ingredients, and is an important natural resource for the development of new drugs. We constructed a T7 phage display library using the cobra (Naja atra) venom gland from the Guangdong Snake Breeding Plant and performed three rounds of screening with TLR4 as the target, randomly selecting monoclonal phage spots for PCR followed by Sanger sequencing. The obtained sequences were subjected to length analysis, molecular docking, solubility prediction, and stability prediction, and a peptide containing 39 amino acids (NA39) was finally screened out. The BLAST results indicated that NA39 was a sequence in RPL19 (Ribosomal Protein L19). After peptide synthesis, the binding ability of NA39 to TLR4 was verified by the surface plasmon resonance (SPR) technique. In this study, a new peptide that can specifically bind TLR4 was successfully screened from the cobra venom gland cDNA library, further demonstrating the effectiveness of phage display technology in the field of drug discovery. [ABSTRACT FROM AUTHOR]

Naja nigricollis Venom (NnV) contains complex toxins that affects various vital systems functions after envenoming. The venom toxins have been reported to induce male reproductive disorders in envenomed rats. This present study explored the ameliorative potential of kaempferol on NnV-induced male reproductive toxicity. Fifty male wistar rats were sorted randomly into five groups (n = 10) for this study. Group 1 were noted as the control, while rats in groups 2 to 5 were injected with LD50 of NnV (1.0 mg/kg bw; i.p.). Group 2 was left untreated post envenomation while group 3 was treated with 0.2 ml of polyvalent antivenom. Groups 4 and 5 were treated with 4 and 8 mg/kg of kaempferol, respectively. NnV caused substantial reduction in concentrations of follicle stimulating hormone, testosterone and luteinizing hormone, while sperm motility, volume and counts significantly (p < 0.05) decreased in envenomed untreated rats. The venom enhanced malondialdehyde levels and substantially decreased glutathione levels, superoxide dismutase and glutathione peroxidase activities in the testes and epididymis of envenomed untreated rats. Additionally, epididymal and testicular myeloperoxidase activity and nitric oxide levels were elevated which substantiated severe morphological defects noticed in the reproductive organs. However, treatment of envenomed rats with kaempferol normalized the reproductive hormones with significant improvement on sperm functional parameters. Elevated inflammatory and oxidative stress biomarkers in testis and epididymis were suppressed post kaempferol treatment. Severe histopathological lesions in the epididymal and testicular tissues were ameliorated in the envenomed treated groups. Results highlights the significance of kaempferol in mitigating reproductive toxicity induced after snakebite envenoming. [ABSTRACT FROM AUTHOR]

Snakebite in India is a severe problem as it causes a mortality rate of 58,000 and a disability rate of 140,000 every year which is the highest among any other country. Antivenom is the primary therapy for snakebite, and its manufacturing techniques have essentially stayed unaltered for over a century. Indian polyvalent antivenom, a scientifically validated medicine for treating the toxic effects of snakebites, is available against the venom of the so-called Big Four snakes namely Spectacled cobra (Naja naja), Saw-scaled viper (Echis carinatus), Russell's viper (Daboia russelli) and the Common krait (Bungarus caeruleus), responsible for majority of the deaths in India. India hosts many other species of snakes, including cobras, kraits, saw-scaled vipers, sea snakes, and pit vipers, responsible for clinically severe envenomation. Neutralization strategy has been applied to access the efficacy of antivenoms, crucial for reducing snake bite deaths and disabilities. This review aims to conduct a systematic review and meta-analysis on the neutralization efficiency of the Polyvalent Antivenom (PAV) and focus on the factors that may contribute to the poor recognition of the antivenom towards the venom toxins. Reports focusing on the investigation of antivenom efficacy were searched and collected from several databases. Preclinical studies that reported the neutralization efficacy of the commercial antivenom against the medically important snakes of India were included. The articles were screened based on the inclusion criteria and 8 studies were shortlisted for meta-analysis. Pooled proportion was calculated for the antivenom efficacy reported by the studies and was found to be statistically significant with a 95% confidence interval. The heterogenicity in the venom toxicity and neutralization potency of the antivenom was evident in the overall estimate (proportion) and individual data. We provide comprehensive evidence on antivenom efficacy against medically important snakes from various parts of India which may aid in identifying the gaps in snake envenomation therapy and the need for novel potentially improved treatment of snakebites. [ABSTRACT FROM AUTHOR]

Naja nivea (N. nivea) is classed as a category one snake by the World Health Organization since its envenomation causes high levels of mortality and disability annually. Despite this, there has been little research into the venom composition of N. nivea, with only one full venom proteome published to date. Our current study separated N. nivea venom using size exclusion chromatography before utilizing a traditional bottom-up proteomics approach to unravel the composition of the venom proteome. As expected by its clinical presentation, N. nivea venom was found to consist mainly of neurotoxins, with three-finger toxins (3FTx), making up 76.01% of the total venom proteome. Additionally, cysteine-rich secretory proteins (CRISPs), vespryns (VESPs), cobra venom factors (CVFs), 5′-nucleotidases (5′NUCs), nerve growth factors (NGFs), phospholipase A2s (PLA2), acetylcholinesterases (AChEs), Kunitz-type serine protease inhibitor (KUN), phosphodiesterases (PDEs), L-amino acid oxidases (LAAOs), hydrolases (HYDs), snake venom metalloproteinases (SVMPs), and snake venom serine protease (SVSP) toxins were also identified in decreasing order of abundance. Interestingly, contrary to previous reports, we find PLA2 toxins in N. nivea venom. This highlights the importance of repeatedly profiling the venom of the same species to account for intra-species variation. Additionally, we report the first evidence of covalent protein complexes in N. nivea venom, which likely contribute to the potency of this venom. [ABSTRACT FROM AUTHOR]

Introduction: Despite improved treatment options for plaque psoriasis within the last decades, some patients still have an inadequate response to treatment. Direct clinical evaluation between therapies used after biologic failure could facilitate physicians' choice of treatment. Methods: COBRA (NCT04533737) was a randomized (1:1), blinded (patient and assessor), 28-week, active-comparator trial conducted in Europe from December 2020 to December 2022. The objective was to compare the efficacy and safety of brodalumab versus guselkumab in adults with moderate-to-severe plaque psoriasis and inadequate response to ustekinumab. Patients received either brodalumab 210 mg or guselkumab 100 mg. The primary [having Psoriasis Area and Severity Index (PASI)-100 response at week 16] and key secondary (time to PASI-100 response) endpoints were tested in a fixed sequence. Results: Due to delays and enrollment challenges, recruitment was terminated with 113 patients enrolled of 240 planned. The proportion of patients having PASI-100 at week 16 for brodalumab was 53.4% compared with 35.9% for guselkumab [odds ratio (OR) 2.05; 95% confidence interval (CI) 0.95, 4.44; p = 0.069]. As this was not statistically significant, the hierarchical testing procedure was stopped. All other secondary PASI endpoints had nominal p-values below 0.05 in favor of brodalumab. In the time to PASI response analyses, brodalumab separated from guselkumab in estimated cumulative incidence of patients achieving a response from week 2 onward, suggesting fast onset of action with brodalumab. Quality of life measures improved in both treatment groups. The safety findings were consistent with the known safety profiles. Conclusions: Brodalumab showed a tendency toward better and earlier effect than guselkumab in patients who had failed ustekinumab. Thus, this trial provides important information in assisting physicians in their choice of therapy for patients who have failed their prior anti-interleukin (IL)-12/23 treatment. Trial Registration: ClinicalTrials.gov identifier NCT04533737. [ABSTRACT FROM AUTHOR]

Effective last-mile (LM) delivery is critical to the efficient functioning of supply chains. In addition to speed and the cost of delivery, environmental and social sustainability are increasingly important factors in last-mile logistics (LML), especially in urban areas. Sustainable solutions such as drones attract special attention from researchers due to their high potential. The future of drone logistics is uncertain due to many barriers. This study analyzes, evaluates and ranks barriers to identify those that most significantly hinder broader drone adoption in LML, and proposes and ranks strategies to overcome them. This type of issue requires the involvement of multiple stakeholders with conflicting goals and interests. Therefore, the study employs a novel hybrid multi-criteria decision-making (MCDM) model that combines fuzzy Delphi-based fuzzy factor relationship (Fuzzy D-FARE) and fuzzy comprehensive distance-based ranking (Fuzzy COBRA) methods. The results indicate that the main obstacle to drone implementation in LM is the lack of aviation regulations. The risks of unauthorized access, data misuse, privacy breaches, and data security represent significant challenges. They are followed by ambiguously defined or burdensome requirements for insurance and liability for drone owners. The main contributions of this study are the establishment of a novel hybrid model, identification and ranking of barriers for broader application of drones in LML, and strategies for overcoming them. [ABSTRACT FROM AUTHOR]

Snake bite is a significant public health concern, particularly in tropical regions. Individuals who are bitten by neurotoxin snake commonly present with ptosis, ophthalmoparesis, muscle weakness, and diminished or absent of deep tendon reflexes. However, accurately determining the occurrence of a snakebite can sometimes be challenging, potentially leading to misdiagnosis. We present the case of a 2-year-old boy with sudden cardiac arrest. Following a brief resuscitation, he had return of spontaneous circulation. Despite normal electroencephalography results, the patient continued to have absence of brainstem reflexes, spontaneous breathing, and movement. Cobra antivenom was promptly initiated based on suspicions of a neurotoxic snakebite, resulting in a rapid recovery of the patient's condition. We proposed that neurotoxin snake envenomation should be considered in patients with sudden cardiac arrest with uncertain cause, particularly in snake endemic areas. [ABSTRACT FROM AUTHOR]

Snake envenomation results in a range of clinical sequelae, and widely used animal-based conventional antivenoms exhibit several limitations including the adverse immunological effects in human snake bite victims. Therefore, human monoclonal anti-snake venom antibodies or fragments can be an alternate therapy for overcoming the existing limitations. We developed venom-neutralizing humanized scFv antibodies and analyzed biochemical mechanisms associated with the inhibition of toxicity. Tomlinson I and J human scFv antibody libraries were screened against Naja naja and Echis carinatus venoms, and seven unique scFv antibodies were obtained. Further, specific toxins of snake venom interacting with each of these scFvs were identified, and phospholipase A2 (PLA2) was found to be prominently captured by the phage-anchored scFv antibodies. Our study indicated PLA2 to be one of the abundant toxins in Naja naja and Echis carinatus venom samples. The scFvs binding to PLA2 were used to perform in vivo survival assay using the mouse model and in vitro toxin inhibition assays. scFv N194, which binds to acidic PLA2, protected 50% of mice treated with Naja naja venom. Significant prolongation of survival time and 16% survival were observed in Echis carinatus venom-challenged mice treated with scFv E113 and scFv E10, respectively. However, a combination comprised of an equal amount of two scFvs, E113 and E10, both interacting with basic PLA2, exhibited synergistically enhanced survival of 33% in Echis carinatus venom-challenged mice. No such synergistically enhanced survival was observed in the case of combinatorial treatment with anti- Naja naja scFvs, N194, and N248. These scFvs demonstrated partial inhibition of venom-induced myotoxicity, and E113 also inhibited hemolysis by 50%, which corroborates the enhanced survival during combinatorial treatment in Echis carinatus venom-challenged mice. • Conventional antivenoms exhibit adverse effects in snakebite victims. • Seven human scFvs antibody fragments against two prominent Indian snake venoms were isolated. • Majority of antibodies were specific to phospholipase A2, which is an abundant toxin in both venoms. • These antibodies partially inhibited venom-induced myotoxicity, and one of them also exhibited partial hemolysis inhibition. • Antibodies developed against PLA2 exhibited partial in vivo venom neutralization. [ABSTRACT FROM AUTHOR]

Standardized use of venous thromboembolism (VTE) risk assessment models (RAMs) in surgical patients has been limited, in part due to the cumbersome workflow addition required to use available models. The COBRA score—capturing cancer diagnosis, (old) age, body mass index, race, and American Society of Anesthesiologists Physical Status score—has been reported as a potentially automatable VTE RAM that circumvents the cumbersome workflow addition that most RAMs represent. We aimed to test the ability of the COBRA model to effectively risk-stratify patients across various populations. Patients were included from the 2014-2019 American College of Surgeons National Surgical Quality Improvement Program (NSQIP) Participant Use Data File for two hospitals, representing colorectal, endocrine, breast, transplant, plastic, and general surgery services. COBRA score was calculated for each patient using preoperative characteristics. We calculated negative predictive value (NPV) for VTE outcomes and compared the COBRA score to NSQIP's expected VTE rate for all patients, between the two hospitals, and between subspecialty service lines. Of the 10,711 patients included, those with COBRA <4 (31%) had projected median VTE rate of 0.21% (interquartile range, 0.09-0.68%; mean, 0.54%). Patients with higher scores (69%) had median rate of 0.88% (0.26-2.07%; 1.46%); relative rate 2.7. The median projected VTE rates for patients identified as low risk were 0.21% and 0.16% and as high risk were 0.87% and 0.89% at hospitals one and 2, respectively. The median projected VTE rates for patients identified as low risk were 0.17%, 0.61%, and 0.08% and as high risk were 0.52%, 1.43%, and 0.18% among general, colorectal, and endocrine surgery patients, respectively. COBRA had NPV of 0.995 and sensitivity of 0.871 as compared to NPV 0.997 and sensitivity 0.857 of the NSQIP model. The COBRA score is concordant with the traditional gold standard NSQIP VTE RAM and demonstrates interhospital and service-specific generalizability, although performance was limited in especially low-risk patients. The model adequately risk-stratifies surgical patients preoperatively, potentially providing clinical decision support for perioperative workflows. [ABSTRACT FROM AUTHOR]

Among the countless images that populate the human imagination, the snake is one of the most powerful. By composing different narratives around the world, it incorporates cosmogonic, eschatological plots, referring to the renewal of time and the transformation of life, as well as being a synthesis of death and the demonstration of human fragility. From the perspective of the snake image, as a multifaceted, ambivalent and plurisemantic symbol, we seek in this work to understand the different narratives about snakes, in a complex of quilombola communities, called "Complexo Quilombola de Mata Cavalo" or "Quilombo de Mata Cavalo", located in the municipality of Nossa Senhora do Livramento, Mato Grosso, Brazil. From a qualitative approach, based on the cartography of the Imaginary (Sato, 2011), interdisciplinary research on the imaginary and symbolism (Durand, 2002); Eliade (1969, 2008), Chevalier and Gheerbrant (2015), among other authors, we will highlight in this work how snakes enter the community's imagination and move from the natural field to the supernatural, which affect the narrators' actions and bring them closer to other universal symbolic perspectives. Whether in the practices of "benzedeiros" and "benzedeiras", in the Judeo-Christian beliefs disseminated in the communities of Mata Cavalo, in the manipulation of animal parts, in the making of medicines, ornaments, etc., snakes continue to be a synthesis of mysteries and stories, permeated by her characteristics and behaviors, which launch her into this borderless field between culture, nature and supernatural. [ABSTRACT FROM AUTHOR]

The document titled "Instrucciones Para la Preparación de Manuscritos en Español" provides guidelines for authors submitting manuscripts to the Southwestern Entomologist journal. The journal accepts manuscripts reporting entomological research conducted in the southwestern United States or Mexico, or studies on arthropods relevant to this region. Routine laboratory or field work reports are not accepted, but reports on experiments with insecticides and other related topics are acceptable. The document also provides instructions on formatting, including font size and style, alignment, and the inclusion of tables and figures. Authors must be members of the Society of Southwestern Entomologist and should submit their manuscripts electronically. The document also mentions the availability of a section for general interest contributions and the acceptance of supplements. The guidelines emphasize the use of proper citation and referencing formats. [Extracted from the article]

This document is a reply to a comment made on an article about optic neuritis caused by snakebites. The authors thank the commenter for their interest and acknowledge that antivenom could be a potential cause of optic neuritis in snakebite victims. The primary aim of the original article was to report the MRI findings in patients with snakebite-induced optic neuritis and emphasize the importance of early diagnosis and treatment. The authors also highlighted the need for further research to understand the underlying mechanisms of optic neuritis in snakebite victims. The authors declare no conflicts of interest. [Extracted from the article]

This document is a comment on a case report published in the journal Toxins. The case report describes two cases of severe optic neuropathy following snakebite envenomation. The patients were treated with antivenom but developed bilateral optic neuropathy several days after the bite. The author of the comment suggests that antivenom may be a cause of the reported symptoms, based on other case reports in the literature. However, the author acknowledges that more research is needed to confirm this relationship and encourages future case reports to provide more detailed information about the type and dosage of antivenom used. [Extracted from the article]

African cobras (Naja species) represent one of the most encountered medically important snakes in Africa. They are classified as African spitting (Afronaja subgenus) and non-spitting cobras (Uraeus and Boulengerina subgenera) with similar and different characteristics. Snake venom toxins including three-finger toxin (3FTx), phospholipase A2 (PLA 2), and snake venom metalloproteinase (SVMP) cause snakebite envenomation leading to morbidity and mortality. The profile of the proteome of African cobra venoms will help to develop safer and more effective antivenoms. The approval of Captopril by the US Food and Drug Administration (FDA) for the treatment of cardiovascular diseases, has led to intensified research towards possible use of venom toxins as therapeutics. In this review, we compare the venom proteome profile of 3 African Naja subgenera. In both Afronaja and Boulengerina subgenera, 3FTx (Afronaja –69.79%; Boulengerina –60.56%) followed by PLA 2 (Afronaja –21.15%; Boulengerina –20.21%) dominated the venoms compared to the Uraeus subgenus dominated by 3FTx (84.55%) with little to no PLA 2 abundance (0.8%). The venom of subgenus Uraeus was distinct from the other two subgenera by the almost total absence of PLA 2 , thus indicating little or no contribution of PLA 2 in the envenomation caused by Uraeus compared to Afronaja and Boulengerina. Furthermore, we report studies on the experimental testing of African cobra venoms and toxins against diseases including anti-cancer properties. [Display omitted] • African spitting and non-spitting cobra venoms cause severe envenomation. • 3FTxs are the most dominant toxin of spitting and non-spitting cobras venoms. • African non-spiting cobra venom of the Uraeus genus constitutes little or no PLA 2. • African cobras have potential therapeutic applications against diseases including cancer. [ABSTRACT FROM AUTHOR]

Patients occasionally present with reports of ocular exposure to fluids from rattlesnakes, claiming or suspecting the substance to be venom. This study set out to evaluate and characterize reported cases of suspected venom-induced ophthalmia in humans. A retrospective review of rattlesnake exposures reported to the Arizona Poison and Drug Information Center over a 24-year period was conducted for ocular exposures. Recorded information included patient demographics, clinical course, laboratory results, and treatments. Documentation regarding interactions between patients and snakes was reviewed by Arizona Poison and Drug Information Center herpetologists to evaluate what substance was expelled from the snake resulting in ocular exposure. Our review of rattlesnake encounters found a total of 26 ocular exposure cases. Patient demographics were largely intentional interactions and involved the male sex. Symptoms ranged from asymptomatic to minor effects with 46.2% managed from home and treated with fluid irrigation. A review of cases by herpetologists concluded the exposure patients commonly experienced was to snake musk. Kinematics of venom expulsion by rattlesnakes conclude the venom gland must be compressed, fangs erected to ≥60o, and fang sheath compressed against the roof of the mouth for venom expulsion. Evidence suggests the chance of venom "spitting" by rattlesnakes is close to zero. Rattlesnakes are documented to forcefully expel airborne malodorous "musk" defensively. An important distinction to remember is musk has a foul odor and is usually colorless, while venom is comparatively odorless and yellow. Rattlesnake venom-induced ophthalmia is a rare event as venom expulsion requires the kinematics of feeding or defensive bites. If the rattlesnake is not in the process of biting or otherwise contacting some other object with its mouth, it is more biologically plausible patients are being exposed to snake musk as a deterrent. Whether it's venom or musk, topical exposure to the eyes should prompt immediate irrigation. [Display omitted] • Rattlesnakes do not "spit" venom as documented in some cobras. • Rattlesnakes may spray venom into the air in the process of biting something, such as striking a screened cage vent. • Rattlesnakes frequently spray malodorous musk a meter or more at antagonists. • Musk has a prominent foul odor while venom is virtually odorless. • Whether it's rattlesnake venom or musk, non-penetrating topical exposure to the eyes should prompt immediate irrigation. [ABSTRACT FROM AUTHOR]

Naja naja snake bite causes thousands of deaths worldwide in a year. N. naja envenomed victims exhibit both local and systemic reactions that potentially lead to death. In clinical practice, pulmonary complications in N. naja envenomation are commonly encountered. However, the molecular mechanisms underlying N. naja venom-induced lung toxicity remain unknown. Here, we reasoned that N. naja venom-induced lung toxicity is prompted by NLRP3 inflammasome and MAPKs activation in mice. Treatment with dimethyl ester of bilirubin (BD1), significantly inhibited the N. naja venom-induced activation of NLRP3 inflammasome and MAPKs both in vivo and in vitro (p < 0.05). Further, BD1 reduced N. naja venom-induced recruitment of inflammatory cells, and hemorrhage in the lung toxicity examined by histopathology. BD1 also diminished N. naja venom-induced local toxicities in paw edema and myotoxicity in mice. Furthermore, BD1 was able to enhance the survival time against N. naja venom-induced mortality in mice. In conclusion, the present data showed that BD1 alleviated N. naja venom-induced lung toxicity by inhibiting NLRP3 inflammasome and MAPKs activation. Small molecule inhibitors that intervene in venom-induced toxicities may have therapeutic applications complementing anti-snake venom. [Display omitted] • Dimethyl ester of bilirubin reversed Naja naja venom induced NLRP3 inflammasome and MAPKs activation. • Dimethyl ester of bilirubin ameliorated Naja naja venom-induced local and systemic toxicities in mice. • Dimethyl ester of bilirubin improved the survival time of Naja naja venom-induced mortality in mice. [ABSTRACT FROM AUTHOR]

This report details a documented case of fatal King cobra (Ophiophagus hannah) envenomation in the Philippines. A 46-year-old woman from a mountainous town in Leyte was bitten on her left thigh by a snake. Despite receiving prompt medical attention, including administration of fluids and oxygen, she went into arrest and succumbed within 2.5 hours of the bite. Inadequate pre-hospital care, including endotracheal intubation and assisted ventilation, highlights a notable gap in emergency medical services. Photographic evidence, verified by a herpetologist, confirmed the involvement of a King cobra, with venom presenting with a swift and lethal systemic effect that led to the patient's demise, despite minimal local manifestations. This incident accentuates the urgent need for accessible, effective antivenom and improved snakebite management protocols in the Philippines. It also calls for heightened awareness and preparedness among pre-hospital healthcare providers and the public, alongside advocating for more research into snakebite envenomation. [Display omitted] • The report introduces a case of fatal King cobra envenomation in the Philippines. • A 46-year-old woman from Leyte experienced a cardiopulmonary arrest within 2.5 hours after being bitten on her left thigh. • Antivenom was unavailable at the time of the incident. • Photographic evidence later confirmed the snake as a King cobra. • This incident points to the need for more research on King cobra bite and improving antivenom access in the Philippines. [ABSTRACT FROM AUTHOR]

Catuneragam nilotica has been used in ethnomedicine to treat snakebite, inflammation, and diarrhea among others. The aim of this research is to isolate, and characterize potential potential phospholipase A 2 (PLA 2) inhibitors from the roots of C. nilotica. The plant material was collected, authenticated, and sequentially extracted using solvents of increasing polarity starting from n-hexane, ethyl acetate, and methanol. The extracts as reported in our previous work, were screened in vitro for their inhibitory activity against PLA 2 enzyme from N. nigricollis venom using acidimetric assay. In line with the bio-activity guided isolation, methanol extract (being the most active) was subjected to chromatographic separation using silica gel and sephadex LH-20 which resulted in the isolation and characterization of scopoletin, and scopolin; the compounds were able to inhibit the hydrolytic actions of PLA2 enzyme with percentage inhibition ranging from 67.82 to 100.00 % and 65.76–93.15 %, respectively while the standard Antisnake Venom (ASV) had 74.96–85.04 % after 10 min incubation at 37 °C. The molecular docking of the compounds against PLA 2 enzyme was performed using Auto Dock Vina while ADME-Tox analysis was evaluated using swissADME and ProTox-II online servers; The findings indicated that both compounds were able to bind to the active site of PLA 2 enzyme with high affinity (−6.5 to −6.2 kcal/mol) and they exhibited favorable drug-likeness and pharmacokinetic properties, and according to toxicity predictions, scopolin was found to be non-toxic (LD 50 of 5000 mg/kg) while scopoletin has a slight chance of being toxic (LD 50 of 3800 mg/kg). In conclusion, the findings of the research revealed that the roots of C. nilotica contains phytoconstituents with anti-PLA 2 enzyme activity and thus, validates the ethnomedicinal claim of the use of the plant as herbal therapy against N. nigricollis envenomation. [Display omitted] • Scopoletin and Scopolin were identified from the roots of Catuneragam nilotica. • Both compounds were able to inhibit the hydrolytic actions of PLA 2 enzyme from Naja nigricollis venom. • The compounds interacted with key amino acid residues such as HIS47 and ASP48 at the active site of PLA 2 with high affinity. • Scopoletin and Scopolin also exhibited favorable drug-like and pharmacokinetics properties. • The roots of C. nilotica contain active phytoconstituents with proven antisnake properties. [ABSTRACT FROM AUTHOR]

Hepatocellular carcinoma and bacterial resistance are major health burdens nowadays. Thus, providing new therapies that overcome that resistance is of great interest, particularly those derived from nature rather than chemotherapeutics to avoid cytotoxicity on normal cells. Venomous animals are among the natural sources that assisted in the discovery of novel therapeutic regimens. L-amino acid oxidase Nh-LAAO (140 kDa), purified from Egyptian Naja haje venom by a successive two-step chromatography protocol, has an optimal pH and temperature of 8 and 37 °C. Under standard assay conditions, Nh-LAAO exhibited the highest specificity toward L-Arg, L-Met and L-Leu, with K m and V max values of 3.5 mM and 10.4 μmol/min/ml, respectively. Among the metal ions, Ca+2, Na+, and K+ ions are activators, whereas Fe+2 inhibited LAAO activity. PMSF and EDTA slightly inhibited the Nh-LAAO activity. In addition, Nh-LAAO showed antibacterial and antifungal activities, particularly against Gentamicin-resistant P. aeruginosa and E. coli strains with MIC of 18 ± 2 μg/ml, as well as F. proliferatum and A. parasiticus among the selected human pathogenic strains. Furthermore, Nh-LAAO exhibited anti-proliferative activity against cancer HepG2 and Huh7 cells with IC50 of 79.37 and 60.11 μg/ml, respectively, with no detectable effect on normal WI-38 cells. Consequently, the apoptosis % of the HepG2 and Huh7 cells were 12 ± 1 and 34.5 ± 2.5 %, respectively, upon Nh-LAAO treatment. Further, the Nh-LAAO arrested the HepG2 and Huh7 cell cycles in the G0/G1 phase. Thus, the powerful selective cytotoxicity of L-amino acid oxidase opens up the possibility as a good candidate for clinical cancer therapy. [Display omitted] • A novel homodimeric L-amino acid oxidase with two subunits of 70 kDa was purified from Egyptian Cobra (Naja haje) venom. • The purified L-amino acid oxidase is optimally active at pH 8 and 37 °C with L-Leucine as a substrate. • Naja haje - L-amino acid oxidase displays anti-bacterial effect against several bacterial strains. • The purified L-amino acid oxidase induces apoptotic activity against hepatic cancer cell lines. • This study offers insight into L-amino acid oxidase purified from Naja haje venom as a promising cancer drug candidate. [ABSTRACT FROM AUTHOR]

Snakebite envenomation often leads to severe visceral injuries, including acute liver injury (ALI). However, the toxicity mechanism remains unclear. Moreover, varespladib can directly inhibit phospholipase A 2 (PLA 2) in snake venom, but its protective effect on snakebite-induced ALI and the mechanism have not been clarified. Previous studies have shown that snake venom PLA 2 leads to neuron cell death via reactive oxygen species (ROS), one of the initial factors related to the mitophagy pathway. The present study group also found that ROS accumulation occurred after Naja atra envenoming. Hematoxylin and eosin (H/E) staining and immunohistochemistry (IHC) were performed to identify the expression of inflammatory factors in the liver tissue, and flow cytometry and immunofluorescence were used to detect ROS levels and mitochondrial function. Immunofluorescence and western blotting were also used for detecting mitophagy pathway-related proteins. The results showed that N. atra bite induced ALI by activating mitophagy and inducing inflammation and that varespladib had a protective effect. Collectively, these results showed the pathological mechanism of ALI caused by N. atra bite and revealed the protective effect of varespladib. [Display omitted] • N.atra venom causes acute liver injury(ALI), which accompanies with ROS generation. • Excess ROS could induce mitophagy. • Varespladib could effectively allevitate ALI caused by N.atra venom. [ABSTRACT FROM AUTHOR]

The frequency of basal cell carcinoma (BCC) cases is putting an increasing strain on dermatopathologists. BCC is the most common type of skin cancer, and its incidence is increasing rapidly worldwide. AI can play a significant role in reducing the time and effort required for BCC diagnostics and thus improve the overall efficiency of the process. To train such an AI system in a fully-supervised fashion however, would require a large amount of pixel-level annotation by already strained dermatopathologists. Therefore, in this study, our primary objective was to develop a weakly-supervised for the identification of basal cell carcinoma (BCC) and the stratification of BCC into low-risk and high-risk categories within histopathology whole-slide images (WSI). We compared Clustering-constrained Attention Multiple instance learning (CLAM) with StreamingCLAM and hypothesized that the latter would be the superior approach. A total of 5147 images were used to train and validate the models, which were subsequently tested on an internal set of 949 images and an external set of 183 images. The labels for training were automatically extracted from free-text pathology reports using a rule-based approach. All data has been made available through the COBRA dataset. The results showed that both the CLAM and StreamingCLAM models achieved high performance for the detection of BCC, with an area under the ROC curve (AUC) of 0.994 and 0.997, respectively, on the internal test set and 0.983 and 0.993 on the external dataset. Furthermore, the models performed well on risk stratification, with AUC values of 0.912 and 0.931, respectively, on the internal set, and 0.851 and 0.883 on the external set. In every single metric the StreamingCLAM model outperformed the CLAM model or is on par. The performance of both models was comparable to that of two pathologists who scored 240 BCC positive slides. Additionally, in the public test set, StreamingCLAM demonstrated a comparable AUC of 0.958, markedly superior to CLAM's 0.803. This difference was statistically significant and emphasized the strength and better adaptability of the StreamingCLAM approach. • Introduces a weakly-supervised, end-to-end BCC detection algorithm. • Algorithm leverages streaming and attention mechanisms. • End-to-end CNN streaming allows efficient learning. • Compared against a state-of-the-art baseline. • Performance benchmarked with two expert pathologists. • Robustness validated on two distinct external datasets. • Achieves top-tier performance in BCC detection and subtyping. • Releases largest public dataset of whole-slide skin images. • Dataset includes 5147 images and publicly available through COBRA. [ABSTRACT FROM AUTHOR]

Here we describe the acute myocardial effects of an elapid (red spitting cobra, Naja pallida) and a viper (western diamondback rattlesnake, Crotalus atrox) venom using an ex vivo heart model. Our results reveal two different pathophysiological trajectories that influence heart function and morphology. While cobra venom causes a drop in contractile force, rattlesnake venom causes enhanced contractility and frequency that coincides with differences in myocellular morphology. This highlights the medical complexity of snake venom-induced cardiotoxicity. [Display omitted] • Highlights_Edit: • Two distinct pathways underlie cardiac (dys)function caused by the venoms of a spitting cobra and a rattlesnake. • Spitting cobra venom decreases contractile force, while rattlesnake venom enhances contractility and frequency. • Venom induced differences in heart cell morphologies coincide with the observed distinct cardiac functions. • Medical complexity underlies snake venom-induced cardiotoxicity. [ABSTRACT FROM AUTHOR]

Three-finger toxins (3FTxs) have traditionally been obtained via venom fractionation of whole venoms from snakes. This method often yields functional toxins, but it can be difficult to obtain pure isoforms, as it is challenging to separate the many different toxins with similar physicochemical properties that generally exist in many venoms. This issue can be circumvented via the use of recombinant expression. However, achieving the correct disulfide bond formation in recombinant toxins is challenging and requires extensive optimization of expression and purification methods to enhance stability and functionality. In this study, we investigated the expression of α-cobratoxin, a well-characterized 3FTx from the monocled cobra (Naja kaouthia), in three different expression systems, namely Escherichia coli BL21 (DE3) cells with the csCyDisCo plasmid, Escherichia coli SHuffle cells, and Komagataella phaffii (formerly known as Pichia pastoris). While none of the tested systems yielded α-cobratoxin identical to the variant isolated from whole venom, the His 6 -tagged α-cobratoxin expressed in K. phaffii exhibited a comparable secondary structure according to circular dichroism spectra and similar binding properties to the α7 subunit of the nicotinic acetylcholine receptor. The findings presented here illustrate the advantages and limitations of the different expression systems and can help guide researchers who wish to express 3FTxs. [Display omitted] • α-cobratoxin produced in bacteria or yeast is not completely identical to the native toxin structure • α-cobratoxin produced in yeast has a similar circular dichroism spectrum as the native toxin and binds the target receptor • Solubilization tags aid in enhancing the expression of soluble protein, but removal negatively affects net yield • E. coli and K. phaffii -expressed α-cobratoxin retains functional binding to its target receptor. • Recombinant toxins can be used to advance antivenom research and the development of toxin-derived therapeutics. [ABSTRACT FROM AUTHOR]

Snake bite envenomation causes tissue damage resulting in acute and chronic inflammatory responses. Inflammasome activation is one of the factors involved in tissue damage in a mouse model of snake envenomation. The present study examines the potency of Indian Big Four snake venoms in the activation of inflammasome and its role in local and systemic tissue toxicity. Among Indian Big Four snake venoms, Naja naja venom activated NLRP3 inflammasome in mouse macrophages. Activation of NLRP3 inflammasome was also observed in mouse foot paw and thigh muscle upon administration of N. naja venom. Intraperitoneal administration of N. naja venom cause systemic lung damage showed activation of NLRP3 inflammasome. Treatment with MCC950, a selective NLRP3 inflammasome inhibitor effectively inhibited N. naja venom-induced activation of caspase-1 and liberation of IL-1β in macrophages. In mice, MCC950 partially inhibited the activation of NLRP3 inflammasome in N. naja venom administered foot paw and thigh muscle. In conclusion, the present data showed that inflammasome is one of the host responses involved in N. naja snake venom-induced toxicities. The inhibition of inflammasome activation will provide new insight into better management of snake bite-induced local tissue damage. Proposed mechanism of NLRP3 inflammasome activation by N. naja venom. LPS priming as a first signal triggers the activation of NLRP3, which leads to the expression of the pro-IL-1β gene and the production of the pro-IL-1β protein. N. naja venom and DAMPs released from muscle tissue due to N. naja venom-induced edema and myotoxicity serve as the second signal for inflammasome activation. These two signals trigger the assembly and activation of the NLRP3 inflammasome complex and the liberate IL-1β cytokine. MCC950 inhibits N. naja venom-induced NLRP3 inflammasome activation by blocking pro-Caspase-1 cleavage into active Caspase-1. [Display omitted] • Naja naja snake venom activates NLRP3 inflammasome complex in macrophages. • Inflammasome pathway is activated in vivo in Naja naja venom-induced edema and myotoxicity in mice. • MCC950 an inhibitor of NLRP3 inflammasome pathway neutralizes Naja naja venom-induced inflammasome activation. [ABSTRACT FROM AUTHOR]

Snake venoms consist of highly biologically active proteins and peptides that are responsible for the lethal physiological effects of snakebite envenomation. In order to guide the development of targeted antivenom strategies, comprehensive understanding of venom compositions and in-depth characterisation of various proteoforms, often not captured by traditional bottom-up proteomic workflows, is necessary. Here, we employ an integrated 'omics' and intact mass spectrometry (MS)-based approach to profile the heterogeneity within the venom of the forest cobra (Naja melanoleuca), adopting different analytical strategies to accommodate for the dynamic molecular mass range of venom proteins present. The venom proteome of N. melanoleuca was catalogued using a venom gland transcriptome-guided bottom-up proteomics approach, revealing a venom consisting of six toxin superfamilies. The subtle diversity present in the venom components was further explored using reversed phase-ultra performance liquid chromatography (RP-UPLC) coupled to intact MS. This approach showed a significant increase in the number of venom proteoforms within various toxin families that were not captured in previous studies. Furthermore, we probed at the higher-order structures of the larger venom proteins using a combination of native MS and mass photometry and revealed significant structural heterogeneity along with extensive post-translational modifications in the form of glycosylation in these larger toxins. Here, we show the diverse structural heterogeneity of snake venom proteins in the venom of N. melanoleuca using an integrated workflow that incorporates analytical strategies that profile snake venom at the proteoform level, complementing traditional venom characterisation approaches. • Forest cobra venom was found to consist of six toxin families, using a transcriptome-guided bottom-up proteomics approach. • Venom proteoform diversity was revealed within various toxin families using RP-UPLC coupled to intact mass spectrometry. • Higher-order structures of larger toxins were profiled using native mass spectrometry and mass photometry. [ABSTRACT FROM AUTHOR]

This article, titled "Expression of Concern: Split inactivated COBRA vaccine elicits protective antibodies against H1N1 and H3N2 influenza viruses," raises concerns about the accuracy and availability of data. The Competing Interests statement has been updated to disclose funding from Sanofi Pasteur, and the involvement of an author affiliated with Sanofi-Pasteur in producing the vaccines. Additionally, the article's Data Availability statement is contradicted by the inability to locate the original raw data files supporting the results. As a result, the article does not comply with the PLOS Data Availability Policy, and an Expression of Concern has been issued by the PLOS ONE Editors. [Extracted from the article]