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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]

Software development effort estimation (SDEE) is a critical activity in developing software. Accurate effort estimation in the early phases of software design life cycle has important effects on the success of software projects. COCOMO (Constructive Cost Model) is a parametric data‐driven SDEE model whose parameters must be calibrated with an organization's local data for accurate estimation. Such data are scarce for most organizations. On the other hand, CoBRA (Cost estimation, Benchmarking, and Risk Assessment) is one of the powerful hybrid methods that need a small number of local historical data for effort estimation. However, data gathering in CoBRA is time‐consuming and costly. To ease the use of CoBRA, in this paper, we design a methodology that extracts CoBRA‐required data from COCOMO datasets. By the proposed method, data collected for COCOMO would be used in CoBRA. Using CoBRA, a more accurate estimation of the required effort would be achieved with fewer number of historical data than what is required to calibrate the COCOMO model. We apply the proposed method on six well‐known public COCOMO datasets and use them in CoBRA. Obtained results depict an increase in the accuracy of estimations in comparison with other existing methods. [ABSTRACT FROM AUTHOR]

Prison violence is a major safety issue for people in prison and staff. In Aotearoa New Zealand, all prison incidents are recorded on the Department of Corrections (Ara Poutama Aotearoa) COBRA database. An analysis of prison violence at the unit level is applied using machine learning to provide a prediction model of prison violence as well as identify important factors associated with violence. The analysis identifies gang affiliation and security class as important predictors of violence in prison. The study also demonstrates the importance of the social environment and spatial scale when conducting an analysis of prison data. [ABSTRACT FROM AUTHOR]

Cardiotoxins, which belong to the family of three-finger toxins, are the main components of cobra venom. They exhibit various types of biological activity, including antimicrobial and cytotoxic against cancer cells. Data on the minimal structural differences between individual toxins are necessary for understanding the molecular mechanisms of their action. This information can be obtained by high-resolution X-ray diffraction analysis. The influence of microgravity on the crystal packing and diffraction quality of crystals of cardiotoxin from cobra Naja naja has been investigated. Cardiotoxin crystals, which were grown on the International Space Station, provided maximally high resolution for the structure of this toxin. Protein crystallized extremely in the hexagonal space group, whereas more than half of crystals grown under laboratory conditions belonged to the orthorhombic system. [ABSTRACT FROM AUTHOR]

Previously, it was shown that the non-conventional toxin WTX from the venom of the cobra Naja kaouthia, when administered intravenously, caused a decrease in blood pressure (BP) and an increase in heart rate (HR) in rats [13]. To identify the site of the toxin molecule responsible for these effects, we studied the influence of synthetic peptide fragments of the WTX on BP and HR in normotensive male Sprague–Dawley rats under general anesthesia induced by Telazol and Xylazine. It was found that peptides corresponding to the WTX central polypeptide loop, stabilized by a disulfide bond, at intravenous injection at concentrations from 0.1 to 1.0 mg/mL caused a dose-dependent decrease in BP, with the HR increasing only in the first 5–10 min after administration. Thus, WTX fragments corresponding to the central polypeptide loop reproduce the decrease in blood pressure caused by the toxin. [ABSTRACT FROM AUTHOR]

The cobra venom factor (CVF) is an unusual non-toxic complement system-activating protein reported in the venom of many family Elapidae snakes, and it is closely related to C3 complement. The archosaurs, including crocodilians, were traditionally considered a completely non-venomous group of animals; however, the identification of some related sequences similar to CVF proteins in the crocodilian genome may trigger important discussions and manage interesting hypotheses. The aim of this work was to identify VF-like protein in Caiman latirostris DNA samples. The results demonstrate the presence of VF-like in C. latirostris and showed a strong homology of VF-like and C3 complement, indicating a probable paralogy between both proteins in crocodilians. Also, these findings provide some potential explanations to justify presence of VF-like protein in crocodilians: one is based on the horizontal transfer of a transposable element from a virus as a vector, and another proposes the horizontal gene transfer by the predator–prey interaction between crocodilians and snakes. Our results are in accordance with investigations that propose the presence of snakes' venom components in non-venomous reptiles. However, more research will be needed to determine if VF-like in crocodilians has a similar function to CVF. [ABSTRACT FROM AUTHOR]

Secure environmental contexts are crucial for hominin interpretation and comparison. The discovery of a Denisovan individual and associated fauna at Tam Ngu Hao 2 (Cobra) Cave, Laos, dating back to 164–131 ka, allows for environmental comparisons between this (sub)tropical site and the Palearctic Denisovan sites of Denisova Cave (Russia) and Baishiya Karst Cave (China). Denisovans from northern latitudes foraged in a mix of forested and open landscapes, including tundra and steppe. Using stable isotope values from the Cobra Cave assemblage, we demonstrate that, despite the presence of nearby canopy forests, the Denisovan individual from Cobra Cave primarily consumed plants and/or animals from open forests and savannah. Using faunal evidence and proxy indicators of climates, results herein highlight a local expansion of rainforest at ~ 130 ka, raising questions about how Denisovans responded to this local climate change. Comparing the diet and habitat of the archaic hominin from Cobra Cave with those of early Homo sapiens from Tam Pà Ling Cave (46–43 ka), Laos, it appears that only our species was able to exploit rainforest resources. [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]

Background: India suffers ~58,000 annual deaths due to snakebites. The 'Big Four' snakes (Russell's viper, Indian cobra, common krait, and saw-scaled viper) that are responsible for most bites cause diverse clinical effects. Delayed treatment increases the risk of serious complications and treatment costs. Although government hospitals offer free treatment for snakebites in India, most patients opt for private healthcare, which is an out-of-pocket expense as they often lack health insurance coverage. This study aims to determine snakebite treatment costs in private tertiary care hospitals in Tamil Nadu, India and identifies the key factors contributing to treatment costs. Methodology/Principal findings: The treatment cost details for 913 snakebite victims were collected from 10 private tertiary care hospitals across Tamil Nadu. The data were classified into hospital, pharmacy, investigation, and laboratory costs, and analysed to determine various factors that contribute to the costs. The results demonstrate that the average treatment costs vary widely for different snakes. The hospital and pharmacy costs are higher than investigation and laboratory costs for all snakebites. Notably, Russell's viper bites cost significantly more than the bites from other snakes. Overall, the type of snake, nature of complications, specialist treatments required, and arrival time to hospitals were identified as some of the key factors for higher treatment costs. Conclusions/Significance: These data demonstrate that ~80% of snakebite patients can be treated with INR 100,000 (~GBP 1000 or USD 1200) or less. This study emphasises the urgent need to improve rural medical care by providing appropriate training for healthcare professionals and essential resources to facilitate early assessment of patients, administer the initial dose of antivenom and refer the patients to tertiary care only when needed. Moreover, the outcome of this study forms a basis for developing appropriate policies to regulate snakebite treatment costs and provide affordable medical insurance for vulnerable communities. Author summary: Snakebite envenoming (SBE) predominantly affects poor communities living in rural areas of developing countries. As SBE induces a wide range of pathological effects in patients, they need a broad spectrum of treatment approaches to tackle those issues. Therefore, antivenom alone is not sufficient to treat SBE patients. In India, most people (including SBE patients) seek treatments from private healthcare settings, although government hospitals provide free treatments. Hence the treatment costs pose a significant burden on the victims and their families leading to severe socioeconomic impacts. To develop better policies to support the clinical management of SBE, it is critical to estimate the costs of SBE treatment in private healthcare settings. In this study, we analysed the treatment costs of 913 snakebite patients who were treated in 10 different private tertiary care hospitals in Tamil Nadu, India, and identified various factors that contribute to treatment costs. For example, snake type, specialist treatments and the time of arrival to the hospital following bites are some of the key factors leading to increased treatment costs. These results form the basis for developing policies to regulate SBE treatment costs, provide health insurance coverage and ensure timely treatment for SBE victims in rural healthcare settings. [ABSTRACT FROM AUTHOR]

Currently, there is an increasing amount of evidence indicating that exosomes and the miRNAs they contain are crucial players in various biological processes. However, the role of exosomes and miRNAs in snake venom during the envenomation process remains largely unknown. In this study, fresh venom from Naja atra of different ages (2-month-old, 1-year-old, and 5-year-old) was collected, and exosomes were isolated through ultracentrifugation. The study found that exosomes with inactivated proteins and enzymes can still cause symptoms similar to cobra envenomation, indicating that substances other than proteins and enzymes in exosomes may also play an essential role in cobra envenomation. Furthermore, the expression profiles of isolated exosome miRNAs were analyzed. The study showed that a large number of miRNAs were co-expressed and abundant in cobra venom exosomes (CV-exosomes) of different ages, including miR-2904, which had high expression abundance and specific sequences. The specific miR-2094 derived from CV-exosomes (CV-exo-miR-2904) was overexpressed both in vitro and in vivo. As a result, CV-exo-miR-2904 induced symptoms similar to cobra envenomation in mice and caused liver damage, demonstrating that it plays a crucial role in cobra envenomation. These results reveal that CV-exosomes and the miRNAs they contain play a significant regulatory role in cobra envenomation. Our findings provide new insights for the treatment of cobra bites and the development of snake venom-based medicines. [ABSTRACT FROM AUTHOR]

Secure environmental contexts are crucial for hominin interpretation and comparison. The discovery of a Denisovan individual and associated fauna at Tam Ngu Hao 2 (Cobra) Cave, Laos, dating back to 164–131 ka, allows for environmental comparisons between this (sub)tropical site and the Palearctic Denisovan sites of Denisova Cave (Russia) and Baishiya Karst Cave (China). Denisovans from northern latitudes foraged in a mix of forested and open landscapes, including tundra and steppe. Using stable isotope values from the Cobra Cave assemblage, we demonstrate that, despite the presence of nearby canopy forests, the Denisovan individual from Cobra Cave primarily consumed plants and/or animals from open forests and savannah. Using faunal evidence and proxy indicators of climates, results herein highlight a local expansion of rainforest at ~ 130 ka, raising questions about how Denisovans responded to this local climate change. Comparing the diet and habitat of the archaic hominin from Cobra Cave with those of early Homo sapiens from Tam Pà Ling Cave (46–43 ka), Laos, it appears that only our species was able to exploit rainforest resources. [ABSTRACT FROM AUTHOR]

The Quaternary vertebrate paleontology of the Brazilian Intertropical Region is well-known, but there are only a few publications on bone ichnology. In this paper, we analyzed trace fossils in 23 bones of the Gruna das Três Cobras, Complexo Suíço, and F3 caves, which are located in Bahia and Rio Grande do Norte states, in the Brazilian Intertropical Region. We discovered gnaw marks assigned to Machichnus and bite marks attributed to Nihilichnus, as well as insect modifications referring to single (Cuniculichnus, Gunnellichnus, Karethraichnus, and Osteocallis) and multiple behaviors (Munitusichnus, and Taotieichnus). Additionally, we proposed the new ichnospecies Nihilichnus sulcatus n. isp. (bite mark) and Cuniculichnus cascudoi n. isp. (insect feeding mark). Regarding the predation/scavenger marks, Protocyon troglodytes was the probable primary modification agent, but some traces may also be attributed to Smilodon populator and Caiman latirostris. Cuniculichnus, Karethraichnus, Munitusichnus, and Osteocallis are attributed to dermestids, whereas termites probably produced Gunnelichnus and Taotieichnus. [ABSTRACT FROM AUTHOR]

Simple Summary: There are millions of snake envenoming cases annually worldwide, which is an important but often neglected tropical disease. The morbidity and mortality from snake bites are not only caused by the venom toxins but also the secondary infection from bacteria from the snake itself or an external source. Therefore, if we do not understand the actual pathogens of infection, the expected treatment efficacy is reduced. In this study, we investigated the microbiota carried in the oral cavity of seven wild venomous snakes in Taiwan using full-length 16S rRNA next-generation sequencing. The results showed that non-pathogenic bacteria and pathogenic bacteria were similar in dominance in the oral cavity of snakes, Gram-negative bacteria were more common than Gram-positive bacteria, and the microbiota were significantly different between the Elapidae and Viperidae families of snakes. Our findings will assist in resolving the diversity of oral microbiomes in snakes and can be applied to the veterinary medicine and clinical therapy of venomous snakebites. A venomous snake's oral cavity may harbor pathogenic microorganisms that cause secondary infection at the wound site after being bitten. We collected oral samples from 37 individuals belonging to seven species of wild venomous snakes in Taiwan, including Naja atra (Na), Bungarus multicinctus (Bm), Protobothrops mucrosquamatus (Pm), Trimeresurus stejnegeri (Ts), Daboia siamensis (Ds), Deinagkistrodon acutus (Da), and alpine Trimeresurus gracilis (Tg). Bacterial species were identified using full-length 16S rRNA amplicon sequencing analysis, and this is the first study using this technique to investigate the oral microbiota of multiple Taiwanese snake species. Up to 1064 bacterial species were identified from the snake's oral cavities, with 24 pathogenic and 24 non-pathogenic species among the most abundant ones. The most abundant oral bacterial species detected in our study were different from those found in previous studies, which varied by snake species, collection sites, sampling tissues, culture dependence, and analysis methods. Multivariate analysis revealed that the oral bacterial species compositions in Na, Bm, and Pm each were significantly different from the other species, whereas those among Ts, Ds, Da, and Tg showed fewer differences. Herein, we reveal the microbial diversity in multiple species of wild snakes and provide potential therapeutic implications regarding empiric antibiotic selection for wildlife medicine and snakebite management. [ABSTRACT FROM AUTHOR]

We characterize the plasma flows generated during the ablation stage of an over-massed exploding planar wire array, fielded on the COBRA pulsed-power facility (1 MA peak current, 250 ns rise time). The planar wire array is designed to provide a driving magnetic field (80 – 100 T) and current per wire distribution (about 60 kA), similar to that in a 10 MA cylindrical exploding wire array fielded on the Z machine. Over-massing the arrays enables continuous plasma ablation over the duration of the experiment without implosion. The requirement to over-mass on the Z machine necessitates wires with diameters of 75 – 100 μ m , which are thicker than wires usually fielded on wire array experiments. To test ablation with thicker wires, we perform a parametric study by varying the initial wire diameter between 33 and 100 μm. The largest wire diameter (100 μm) array exhibits early closure of the cathode-wire gap, while the gap remains open over the duration of the experiment for wire diameters between 33 and 75 μm. Laser plasma interferometry and time-gated extreme-ultraviolet (XUV) imaging are used to probe the plasma flows ablating from the wires. The plasma flows from the wires converge to generate a pinch, which appears as a fast-moving (V ≈ 100 kms − 1 ) column of increased plasma density ( n ¯ e ≈ 2 × 10 18 cm − 3 ) and strong XUV emission. Finally, we compare the results with three-dimensional resistive-magnetohydrodynamic (MHD) simulations performed using the code GORGON, the results of which reproduce the dynamics of the experiment reasonably well. [ABSTRACT FROM AUTHOR]

Snake venoms constitute a complex, rapidly evolving trait, whose composition varies between and within populations depending on geographical location, age and preys (diets). These factors have determined the adaptive evolution for predatory success and link venom heterogeneity with prey specificity. Moreover, understanding the evolutionary drivers of animal venoms has streamlined the biodiscovery of venom-derived compounds as drug candidates in biomedicine and biotechnology. The king cobra (Ophiophagus hannah; Cantor, 1836) is distributed in diverse habitats, forming independent populations, which confer differing scale markings, including between hatchlings and adults. Furthermore, king cobra venoms possess unique cytotoxic properties that are used as a defensive trait, but their toxins may also have utility as promising anticancer-agent candidates. However, the impact of geographical distribution and age on these potential venom applications has been typically neglected. In this study, we hypothesised that ontogenetic venom variation accompanies the morphological distinction between hatchlings and adults. We used non-transformed neonatal foreskin (NFF) fibroblasts to examine and compare the variability of venom cytotoxicity between adult captive breeding pairs from Malaysian and Chinese lineages, along with that of their progeny upon hatching. In parallel, we assessed the anticancer potential of these venoms in human-melanoma-patient-derived cells (MM96L). We found that in a geographical distribution and gender-independent manner, venoms from hatchlings were significantly less cytotoxic than those from adults (NFF; ~Log EC50: 0.5–0.6 vs. 0.2–0.35 mg/mL). This is consistent with neonates occupying a semifossorial habitat, while adults inhabit more above-ground habitats and are therefore more conspicuous to potential predators. We also observed that Malaysian venoms exhibited a slightly higher cytotoxicity than those from the Chinese cobra cohorts (NFF; Log EC50: 0.1–0.3 vs. 0.3–0.4 mg/mL), which is consistent with Malaysian king cobras being more strongly aposematically marked. These variations are therefore suggestive of differential anti-predator strategies associated with the occupation of distinct niches. However, all cobra venoms were similarly cytotoxic in both melanoma cells and fibroblasts, limiting their potential medical applications in their native forms. [ABSTRACT FROM AUTHOR]

Naja atra bite is one of the most common severe snakebites in emergency departments. Unfortunately, the pathophysiological changes caused by Naja atra bite are unclear due to the lack of good animal models. In this study, an animal model of Naja atra bite in Guangxi Bama miniature pigs was established by intramuscular injection at 2 mg/kg of Naja atra venom, and serum metabolites were systematically analyzed using untargeted metabolomic and targeted metabolomic approaches. Untargeted metabolomic analysis revealed that 5045 chromatographic peaks were obtained in ESI+ and 3871 chromatographic peaks were obtained in ESI-. Screening in ESI+ modes and ESI- modes identified 22 and 36 differential metabolites compared to controls. The presence of 8 core metabolites of glutamine, arginine, proline, leucine, phenylalanine, inosine, thymidine and hippuric acid in the process of Naja atra bite was verified by targeted metabolomics significant difference (P<0.05). At the same time, during the verification process of the serum clinical samples with Naja atra bite, we found that the contents of three metabolites of proline, phenylalanine and inosine in the serum of the patients were significantly different from those of the normal human serum (P<0.05). By conducting functional analysis of core and metabolic pathway analysis, we revealed a potential correlation between changes in key metabolites after the Naja atra bite and the resulting pathophysiological alterations, and our research aims to establish a theoretical foundation for the prompt diagnosis and treatment of Naja atra bite. Author summary: Naja atra bite is one of the most serious tropical and subtropical diseases, and the pathophysiological mechanism of injury caused by Naja atra bite is still not clear, meanwhile, large animal models of Naja atra bite are rarely reported. In this study, a Naja atra bite Bama miniature pig model was constructed and evaluated. In addition, 22 and 36 metabolites were found to be significantly altered in ESI+ and ESI- mode by untargeted metabolomics assay, respectively. More interestingly, 8 of them co-occurred in both ESI+ and ESI- modes. We quantified these 8 metabolites absolutely by targeted metabolomics and validated them in the serum of Naja atra bite patients, which showed the same trend as the untargeted metabolomics results. We also compared the differences in the levels of these 8 metabolites in the serum of the Naja atra bite patient group and the normal human group, we found decreased expression of inosine, proline and phenylalanine. Based on this, we compared the blood metabolic profiles with those of Bungarus multicinctus and Trimeresurus stejnegeri, and finally we confirmed that inosine, proline and phenylalanine are likely to be the key metabolites in Naja atra bite. [ABSTRACT FROM AUTHOR]

Background: Snakebite envenoming is a well-known medical emergency in the Terai of Nepal in particular. However, there is an epidemiological knowledge gap. The news media data available online provide substantial information on envenomings. Assessing this information can be a pristine approach for understanding snakebite epidemiology and conducting knowledge-based interventions. We firstly analyzed news media-reported quantitative information on conditions under which bites occur, treatment-seeking behavior of victims, and outcomes of snakebite envenomings in Nepal. Methodology/Principal findings: We analyzed 308 Nepalese snakebite envenomed cases reported in 199 news media articles published between 2010 and 2022 using descriptive statistics, Wilcoxon, and Chi-square tests to know why and how victims were bitten, their treatment-seeking behavior, and the outcomes. These envenomated cases known with substantial information represented 48 districts (mostly located in the Terai region) of Nepal. These envenomings mostly occurred in residential areas affecting children. Generally, envenomings among males and females were not significantly different. But, in residential areas, females were more envenomed than males. Further, victims' extremities were often exposed to venomous snakebites while their active status and these episodes often occurred at night while victims were passive during snakebites indoors and immediate surroundings of houses. Snakebite deaths were less among referred than non-referred cases, males than females, and while active than passive conditions of victims. Conclusion/Significance: The most of reported envenomed patients were children, and most envenomings were due to cobra bites. Consultation with traditional healers complicated snakebite management. In most cases, deaths that occur without medical interventions are a severe snakebite consequence in Nepal. Further, several deaths in urban areas and mountains and higher hills of Nepal suggest immediate need of snakebite management interventions in the most affected districts. Therefore, there is an urgent need to immediately admit Nepalese snakebite victims to nearby snakebite treatment centers without adopting non-recommended prehospital interventions. The strategies for preventing snakebite and controlling venom effects should also include hilly and mountain districts where snakebite-associated deaths are reported. Author summary: Worldwide, 1.8 to 2.7 million snakebite envenomings occur annually resulting in 81,000 to 138,000 deaths. Annually, 1,000 to 3,225 deaths occur in Nepal. However, there is an epidemiological knowledge gap. Since news media data are typically available in near real-time, the news media data can be analyzed to understand the epidemiological situations in Nepal like poor resource country. Therefore, we analyzed news media-reported snakebite envenomings that occurred in the last 13 years. The median age of envenomed patients was 19 y. People were envenomated mainly during natural human activities such as sleeping. Cobras, kraits, Russell's Viper, Mountain Pitviper, and Green Pitvipers were involved in snakebites. Overall, three-fourth of victims who consulted traditional healers died. Forty-four cases being referred to higher centers were died on the way to higher healthcare centers or in healthcare institutions during treatment. Dependency on traditional snakebite healers, the long distance between snakebite localities and snakebite treatment centers, and snakebite occurred at night were the major barriers to accessing healthcare facilities by people bitten by snakes in Nepal. In this country, people inhabiting 48 districts are at risk of venomous snakebites. [ABSTRACT FROM AUTHOR]

Background: Envenomations by African snakes represent a high burden in the sub-Sahara region. The design and fabrication of polyspecific antivenoms with a broader effectiveness, specially tailored for its use in sub-Saharan Africa, require a better understanding of the immunological features of different Naja spp. venoms of highest medical impact in Africa; and to select the most appropriate antigen combinations to generate antivenoms of wider neutralizing scope. Methodology/principal findings: Rabbit-derived monospecific antisera were raised against the venoms of five spitting cobras and six non-spitting cobras. The effects of immunization in the animal model were assessed, as well as the development of antibody titers, as proved by immunochemical assays and neutralization of lethal, phospholipase A2 and dermonecrotic activities. By the end of the immunization schedule, the immunized rabbits showed normal values of all hematological parameters, and no muscle tissue damage was evidenced, although alterations in aspartate aminotransferase (AST) and alkaline phosphatase (ALP) suggested a degree of hepatic damage caused mainly by spitting cobra venoms. Immunologic analyses revealed a considerable extent of cross-reactivity of monospecific antisera against heterologous venoms within the spitting and no-spitting cobras, yet some antisera showed more extensive cross-reactivity than others. The antisera with the widest coverage were those of anti-Naja ashei and anti-N. nigricollis for the spitting cobras, and anti-N. haje and anti-N. senegalensis for the non-spitting cobras. Conclusions/significance: The methods and study design followed provide a rationale for the selection of the best combination of venoms for generating antivenoms of high cross-reactivity against cobra venoms in sub-Saharan Africa. Results suggest that venoms from N. ashei, N. nigricollis within the spitting cobras, and N. haje and N. senegalensis within the non-spitting cobras, generate antisera with a broader cross-reactivity. These experimental results should be translated to larger animal models used in antivenom elaboration to assess whether these predictions are reproduced. Author summary: The African elapids pose a major potential to induce clinically important envenomations and severe sequelae after a snakebite accident. Within the genus Naja, different radiation events have occurred rendering the African cobras with diverse features in their venom composition. Commonly divided into spitting and non-spitting cobras, their venom is characterized as either cytotoxic or neurotoxic, respectively, based on the pathophysiology of envenomation. In this study, an experimental protocol was developed based on the generation of monospecific antisera in rabbits immunized with venoms of spitting and non-spitting African cobras. Cross-reactivity of monospecific antisera was assessed by immunochemical analyses and by neutralization of the lethal, phospholipase A2 (PLA2) and dermonecrotic activities. Results showed a large extent of intrageneric cross-reactivity by all antisera, yet few exceptions were noticed. The venoms that generated antisera with the highest cross-neutralization ability among the spitting cobras were Naja nigricollis and N. ashei, whereas for the non-spitting cobras were N. haje and N. senegalensis. These results provide evidence for the selection of the best combination of immunogens for antivenom production with the widest neutralization scope within the African cobras. Additionally, the results obtained in the rabbit model should be translated to a larger animal model used for antivenom fabrication to assess whether these predictions are confirmed. [ABSTRACT FROM AUTHOR]

Approximate 70% of cobra venom is composed of cytotoxin (CTX), which is responsible for the dermonecrotic symptoms of cobra envenomation. However, CTX is generally low in immunogenicity, and the antivenom is ineffective in attenuating its in vivo toxicity. Furthermore, little is known about its epitope properties for empirical antivenom therapy. This study aimed to determine the epitope sequences of CTX using the immunoinformatic analyses and epitope-omics profiling. A conserved CTX was used in this study to determine its T-cell and B-cell epitope sequences using immunoinformatic tools and molecular docking simulation with different Human Leukocyte Antigens (HLAs). The potential T-cell and B-cell epitopes were 'KLVPLFY,' 'CPAGKNLCY,' 'MFMVSTPTK,' and 'DVCPKNSLL.' Molecular docking simulations disclosed that the HLA-B62 supertype exhibited the greatest binding affinity towards cobra venom cytotoxin. The namely L7, G18, K19, N20, M25, K33, V43, C44, K46, N47, and S48 of CTX exhibited prominent intermolecular interactions with HLA-B62. The multi-enzymatic-limited-digestion/liquid chromatography-mass spectrometry (MELD/LC–MS) also revealed three potential epitope sequences as 'LVPLFYK,' 'MFMVS,' and 'TVPVKR'. From different epitope mapping approaches, we concluded four potential epitope sites of CTX as 'KLVPLFYK', 'AGKNL', 'MFMVSTPKVPV' and 'DVCPKNSLL'. Site-directed mutagenesis of these epitopes confirmed their locations at the functional loops of CTX. These epitope sequences are crucial to CTX's structural folding and cytotoxicity. The results concluded the epitopes that resided within the functional loops constituted potential targets to fabricate synthetic epitopes for CTX-targeted antivenom production. [ABSTRACT FROM AUTHOR]

The world-famous markets of Marrakech, also known in Arabic as souks, harbor a vast diversity of reptiles that are sold for medicinal/magic/pet purposes or used for snake charming. This unique epidemiological context has never been studied considering the interactions of humans, reptiles, and zoonotic pathogens. Thus, the aim of this study was to identify the parasites and pathogens present in blood and feces associated with handled reptiles in the markets of Marrakech to assess the risk of zoonotic transmission within the reptile-human interface. Privately owned reptiles (n = 118), coming from vendors or snake charmers, were examined and blood and feces sampled. DNA was extracted and molecular screening (cPCR, nPCR, qPCR, dqPCR) was performed aiming to identify potentially zoonotic pathogens (i.e., Anaplasma/Ehrlichia spp., Rickettsia spp., Borrelia burgdorferi sensu lato, Coxiella burnetii, Babesia/Theileria spp., Cryptosporidium spp., Giardia spp., Leishmania spp., Cestoda). Overall, 28.9% (34/118) of reptiles were positive for at least one pathogen. In blood, Anaplasma spp. were detected in four snakes, with two Montpellier snakes positive for Anaplasma phagocytophilum, while Rickettsia spp. were detected in one Mediterranean chameleon and four puff adders. Leishmania tarentolae was molecularly detected in a Mediterranean chameleon and a Montpellier snake. In feces, the cox1 gene generated a myriad of sequences for nematodes, cestodes, fungi and bacteria. Importantly, Proteus vulgaris was identified from a Mediterranean chameleon. Cryptosporidium spp. nPCR yielded a positive sample (i.e., Cryptosporidium sp. apodemus genotype I) from a Moroccan worm lizard, as well as for bacteria such as Pseudomonas aeruginosa in an Egyptian cobra, and Morganella morganii from a puff adder. Results from this study demonstrated the risk of zoonotic transmission of microorganisms and parasites present in blood and feces from reptiles that are brought to the souks in Marrakech, Morocco, to be sold for medicinal purposes or used for snake charming, being in direct and straight contact with humans. Author summary: The world-famous Marrakech souks (markets) are a unique and scarcely studied context where reptiles are in constant contact with humans and used for traditional medicine or for the ancient craft of snake charming. Hence, we aimed to identify the pathogens present in blood and feces of reptiles kept in the markets of Marrakech to assess the zoonotic risk. Animals from vendors or snake charmers were sampled. DNA was extracted and molecularly screened to identify potentially zoonotic pathogens. Overall, 28.9% of reptiles were positive for at least one pathogen. Importantly, we detected Anaplasma spp. from four snakes, Rickettsia spp. from Mediterranean chameleons and puff adders, and Leishmania tarentolae from snakes and chameleons. On the other hand, zoonotic bacteria such Proteus vulgaris were identified from a Mediterranean chameleon, Pseudomonas aeruginosa from Egyptian cobra, and Morganella morganii from puff adder. Thus, reptiles that are kept in the souks of Marrakech may play a role in the zoonotic transmission of pathogens and parasites, given the constant contact with humans. [ABSTRACT FROM AUTHOR]

Because of the high incidence of Pseudomonas aeruginosa biofilms-related nosocomial infections, venoms from common Thai snakes were tested. Although venoms from king cobra (Ophiophagus hannah; OH) and green pit viper (Trimeresurus albolabris) showed the broadest antibacterial spectrum, OH venom demonstrated more profound anti-biofilm activities against P. aeruginosa. Additionally, purified l-amino acid oxidase from OH venom (OH-LAAO), using a three-step chromatography and protein identification, reduced biofilm mass as indicated by the downregulation of several genes, including the genes for biofilm synthesis (algD and pslB) and biofilm regulators (algU, gacA, and siaD). Moreover, OH-LAAO disrupted Pseudomonas-preformed biofilms via upregulation of several genes for biofilm dispersion (nbdA, bdlA, and dipA) and biofilm degradation (endA and pslG), resulting in a reduction of the biofilm biomass. Due to the antimicrobial effects and anti-biofilm activities (reduced production plus increased dispersion) neutralized by catalase, a hydrogen peroxide (H2O2)-degrading enzyme, the enhanced H2O2 by OH venom might be one of the anti-biofilm mechanisms. Hence, OH-LAAO was proposed as a novel agent against Pseudomonas biofilms for either treatment or prevention. More studies are interesting. [ABSTRACT FROM AUTHOR]

Background: Snakebite envenoming is a global health problem, mostly in tropical and subtropical countries. Bangladesh is a subtropical country facing thousands of snakebite envenoming death every year. Knowledge of the distribution of venomous snakes is necessary to identify snakebite-prone areas, develop strategies for prevention and management, and reduce venom-induced mortalities and morbidities. Methods: An integrated effort of direct observations of snakes, qualitative analysis of museum specimen records, clinical records of snakebites, and scholarly literature records were used to understand the pattern of distribution of venomous snakes in Bangladesh. Results: We enlist 65 venomous snake species from Bangladesh and present detailed documentation on their distribution pattern. However, only nine species were considered medically relevant species because of their venom potentiality to kill humans and available clinical records of envenomation. The distribution pattern of those species divides the country into two major portions. A portion consists of northern, northwestern, and western parts of the country, which are habitats of Naja naja, Bungarus caeruleus, and B. lividus. Another portion comprised of the northeastern, southeastern, and southern parts of the country provides habitats for N. kaouthia, B. niger, and Trimeresurus erythrurus. However, Daboia. russelii had a different distribution pattern along the bank of the Padma and Meghna, and some coastal districts. Moreover, B. walli had a scattered distribution over the country. The knowledge of this ecological distribution of venomous snakes across the country bears a significant practical effect on clinical management of snakebite. Treating physicians can have a better understanding of possible offending snake species using this knowledge and the clinical syndromes produced by venoms. Conclusion: Pattern of distribution of medically relevant venomous snakes in Bangladesh approaches to initiate concise and specific bite management strategies for two distinct distributional regions of the country. Moreover, the distribution of D. russelii and B. walli demands specific strategies for bite management. [ABSTRACT FROM AUTHOR]

This study investigated the clinical characteristics, treatments, and outcomes of envenomation involving cobra species in Thailand (Naja kaouthia, Naja siamensis, and Naja sumatrana). Data of patients who had been bitten by a cobra or inoculated via the eyes/skin in 2018–2021 were obtained from the Ramathibodi Poison Center. There were 1045 patients admitted during the 4-year study period (bite, n = 539; ocular/dermal inoculation, n = 506). Almost all patients with ocular/dermal inoculation had eye involvement and ocular injuries, but none had neurological effects. Most of the patients bitten by a cobra had local effects (69.0%) and neurological signs and symptoms (55.7%). The median interval between the bite and the onset of neurological symptoms was 1 h (range, 10 min to 24 h). Accordingly, patients should be observed closely in hospitals for at least 24 h after a bite. Intubation with ventilator support was required in 45.5% of patients and for a median duration of 1.1 days. Antivenom was administered in 63.5% of cases. There were nine deaths, most of which resulted from severe infection. Neurological effects and intubation were significantly more common after a monocled cobra bite than after a spitting cobra bite. The administration of antivenom with good supportive care, including the appropriate management of complications, especially wound infection, might decrease fatality. [ABSTRACT FROM AUTHOR]

Introduction: In Taiwan, six venomous snake species with medical importance have been found; however, long-term epidemiological data of snakebite envenomation (SBE) is lacking. This study aimed to explore the epidemiology of SBE based on the distribution and use of different antivenoms in different parts of Taiwan to facilitate the development of prevention strategies and resource allocation. Methods and results: This retrospective study was conducted using the Taiwan National Health Insurance Research Database from 2002 to 2014. A total of 12,542 patients were treated with antivenoms. The directly standardized cumulative incidence was 3.6 cases per 100,000 individuals based on the 2000 World Standard Population. The incidence of SBEs peaked in the summer (35.9%). The relative risk (RR) of male patients versus female patients was 2.5 (p < 0.0001). The RRs of patients aged 18–64 and ≥65 years versus those aged <18 years were 6.0 (p < 0.0001) and 14.3 (p < 0.0001), respectively. Furthermore, the RR of eastern Taiwan versus northern Taiwan was 6.8 (p < 0.0001). The RR of agricultural workers versus laborers was 5.5 (p < 0.0001). Compared with patients envenomed by Trimeresurus stejnegeri stejnegeri or Protobothrops mucrosquamatus, those envenomed by Naja atra or Bungarus multicinctus multicinctus were more likely to occur in central (adjusted odds ratio [aOR] = 2.6, p < 0.0001) or southern (aOR = 3.2, p < 0.0001) Taiwan, but less frequently among agricultural workers (aOR = 0.6, p < 0.0001). The overall case-fatality rate was 0.11%. Conclusions: Among Asian countries, Taiwan had low incidence and case-fatality rates of SBE. Risk factors included male gender, old age, summer season, being in eastern Taiwan, and being an agricultural worker. Differences of the epidemiological findings between snake species should be focused on when developing strategies for snakebite prevention. Author summary: SBE is a neglected tropical disease of highest priority to be aware of, and its epidemiology varies widely worldwide. This study explored the nationwide and long-term epidemiology of SBE based on the use of snake antivenom in Taiwan, including its incidence, risk factors, complications, and management. This could facilitate the development of preventive strategies and resource allocation. In Taiwan, six medically important venomous snake species were found; however, majority (73.3%) of the envenomation events were associated with T. s. stejnegeri (bamboo pit viper) and P. mucrosquamatus (Taiwan habu) and some (17.1%) with N. atra (Chinese cobra) and B. m. multicinctus (many-banded krait). The risks varied depending on the season, sex, age, geographic region, and occupation. Compared with patients envenomed by crotalines, those envenomed by elapids were more likely to undergo surgery due to serious wound complications caused by N. atra or endotracheal intubation due to respiratory failure caused by B. m. multicinctus. The low case-fatality rate observed might be due to easy access to modern medicine, immediate availability of antivenoms in healthcare facilities, few adverse effects of antivenoms, and prompt surgical intervention. The lack of highly venomous snakes with highly potent venom is a leading factor in low case fatality. [ABSTRACT FROM AUTHOR]

The socioeconomic impact of snakebites in India is largely attributed to a subset of snake species commonly known as the 'big four'. However, envenoming by a range of other clinically important yet neglected snakes, a.k.a. the 'neglected many', also adds to this burden. The current approach of treating bites from these snakes with the 'big four' polyvalent antivenom is ineffective. While the medical significance of various species of cobras, saw-scaled vipers, and kraits is well-established, the clinical impact of pit vipers from regions such as the Western Ghats, northeastern India, and the Andaman and Nicobar Islands remains poorly understood. Amongst the many species of snakes found in the Western Ghats, the hump-nosed (Hypnale hypnale), Malabar (Craspedocephalus malabaricus), and bamboo (Craspedocephalus gramineus) pit vipers can potentially inflict severe envenoming. To evaluate the severity of toxicity inflicted by these snakes, we characterised their venom composition, biochemical and pharmacological activities, and toxicity- and morbidity-inducing potentials, including their ability to damage kidneys. Our findings highlight the therapeutic inadequacies of the Indian and Sri Lankan polyvalent antivenoms in neutralising the local and systemic toxicity resulting from pit viper envenomings. [ABSTRACT FROM AUTHOR]

Cardiotoxins (CaTx) of the three-finger toxin family are one of the main components of cobra venoms. Depending on the structure of the N-terminal or the central polypeptide loop, they are classified into either group I and II or P- and S-types, respectively, and toxins of different groups or types interact with lipid membranes variably. While their main target in the organism is the cardiovascular system, there is no data on the effects of CaTxs from different groups or types on cardiomyocytes. To evaluate these effects, a fluorescence measurement of intracellular Ca2+ concentration and an assessment of the rat cardiomyocytes' shape were used. The obtained results showed that CaTxs of group I containing two adjacent proline residues in the N-terminal loop were less toxic to cardiomyocytes than group II toxins and that CaTxs of S-type were less active than P-type ones. The highest activity was observed for Naja oxiana cobra cardiotoxin 2, which is of P-type and belongs to group II. For the first time, the effects of CaTxs of different groups and types on the cardiomyocytes were studied, and the data obtained showed that the CaTx toxicity to cardiomyocytes depends on the structures both of the N-terminal and central polypeptide loops. [ABSTRACT FROM AUTHOR]