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Conotoxins are small and highly potent neurotoxic peptides derived from the venom of marine cone snails which have captured the interest of the scientific community due to their pharmacological potential. These toxins display significant sequence and structure diversity, which results in a wide range of specificities for several different ion channels and receptors. Despite the recognized importance of these compounds, our ability to determine their binding targets and toxicities remains a significant challenge. Predicting the target receptors of conotoxins, based solely on their amino acid sequence, remains a challenge due to the intricate relationships between structure, function, target specificity, and the significant conformational heterogeneity observed in conotoxins with the same primary sequence. We have previously demonstrated that the inclusion of post-translational modifications, collisional cross sections values, and other structural features, when added to the standard primary sequence features, improves the prediction accuracy of conotoxins against non-toxic and other toxic peptides across varied datasets and several different commonly used machine learning classifiers. Here, we present the effects of these features on conotoxin class and molecular target predictions, in particular, predicting conotoxins that bind to nicotinic acetylcholine receptors (nAChRs). We also demonstrate the use of the Synthetic Minority Oversampling Technique (SMOTE)-Tomek in balancing the datasets while simultaneously making the different classes more distinct by reducing the number of ambiguous samples which nearly overlap between the classes. In predicting the alpha, mu, and omega conotoxin classes, the SMOTE-Tomek PCA PLR model, using the combination of the SS and P feature sets establishes the best performance with an overall accuracy (OA) of 95.95%, with an average accuracy (AA) of 93.04%, and an f1 score of 0.959. Using this model, we obtained sensitivities of 98.98%, 89.66%, and 90.48% when predicting alpha, mu, and omega conotoxin classes, respectively. Similarly, in predicting conotoxins that bind to nAChRs, the SMOTE-Tomek PCA SVM model, which used the collisional cross sections (CCSs) and the P feature sets, demonstrated the highest performance with 91.3% OA, 91.32% AA, and an f1 score of 0.9131. The sensitivity when predicting conotoxins that bind to nAChRs is 91.46% with a 91.18% sensitivity when predicting conotoxins that do not bind to nAChRs. [ABSTRACT FROM AUTHOR]

Mollusk-hunting (molluscivorous) cone snails belong to a monophyletic group in Conus , a genus of venomous marine snails. The molluscivorous lineage evolved from ancestral worm-hunting (vermivorous) snails ∼18 Ma. To enable the shift to a molluscivorous lifestyle, molluscivorous cone snails must solve biological problems encountered when hunting other gastropods, namely: (i) preventing prey escape and (ii) overcoming the formidable defense of the prey in the form of the molluscan shell, a problem unique to molluscivorous Conus. Here, we show that χ-conotoxins, peptides exclusively expressed in the venoms of molluscivorous Conus , provide solutions to the above problems. Injecting χ-conotoxins into the gastropod mollusk Aplysia californica results in impaired locomotion and uncoordinated hyperactivity. Impaired locomotion impedes escape, and a hyperactive snail will likely emerge from its shell, negating the protection the shell provides. Thus, χ-conotoxins are an evolutionary innovation that accompanied the emergence of molluscivory in Conus and provide solutions to problems posed by hunting other snails. [ABSTRACT FROM AUTHOR]

This article discusses the genome assembly and characterization of Chlorogloeopsis sp. ULAP02, a cyanobacterium from the Philippines, and the identification of biosynthetic gene clusters (BGCs) responsible for producing bioactive compounds. The study reveals the organism's genome and the potential therapeutic applications of its compounds. Another article explores the genome of Chlorogloeopsis sp. ULAP02 and identifies various BGCs associated with antimicrobial properties. Additionally, a document provides a list of references for scientific articles on microbiology and biochemistry, covering topics such as microorganism classification, protein engineering, and natural product biosynthesis. These resources offer valuable information for researchers in these fields. [Extracted from the article]

Genetic generalized epilepsies (GGE) exhibit widespread morphometric alterations in the subcortical structures. Subcortical structures are essential for understanding GGE pathophysiology, but their fine-grained morphological diversity has yet to be comprehensively investigated. Furthermore, the relationships between macroscale morphological disturbances and microscale molecular chemoarchitectures are unclear. High-resolution structural images were acquired from patients with GGE (n = 97) and sex- and age-matched healthy controls (HCs, n = 184). Individual measurements of surface shape features (thickness and surface area) of seven bilateral subcortical structures were quantified. The patients and HCs were then compared vertex-wise, and shape anomalies were co-located with brain neurotransmitter profiles. We found widespread morphological alterations in GGE and prominent disruptions in the thalamus, putamen, and hippocampus. Shape area dilations were observed in the bilateral ventral, medial, and right dorsal thalamus, as well as the bilateral lateral putamen. We found that the shape area deviation pattern was spatially correlated with the norepinephrine transporter and nicotinic acetylcholine (Ach) receptor (α4β2) profiles, but a distinct association was seen in the muscarinic Ach receptor (M1). The findings provided a comprehensive picture of subcortical morphological disruptions in GGE, and further characterized the associated molecular mechanisms. This information may increase our understanding of the pathophysiology of GGE. Genetic generalized epilepsies (GGE)exhibited widespread shape alterations in subcortical structures and these morphological anomalies linked to specific neurotransmitter profiles [ABSTRACT FROM AUTHOR]

Background and Purpose: The analgesic action of paracetamol involves KV7 channels, and its metabolite N‐acetyl‐p‐benzo quinone imine (NAPQI), a cysteine modifying reagent, was shown to increase currents through such channels in nociceptors. Modification of cysteine residues by N‐ethylmaleimide, H2O2, or nitric oxide has been found to modulate currents through KV7 channels. The study aims to identify whether, and if so which, cysteine residues in neuronal KV7 channels might be responsible for the effects of NAPQI. Experimental Approach: To address this question, we used a combination of perforated patch‐clamp recordings, site‐directed mutagenesis, and mass spectrometry applied to recombinant KV7.1 to KV7.5 channels. Key Results: Currents through the cardiac subtype KV7.1 were reduced by NAPQI. Currents through all other subtypes were increased, either by an isolated shift of the channel voltage dependence to more negative values (KV7.3) or by such a shift combined with increased maximal current levels (KV7.2, KV7.4, KV7.5). A stretch of three cysteine residues in the S2‐S3 linker region of KV7.2 was necessary and sufficient to mediate these effects. Conclusion and Implication: The paracetamol metabolite N‐acetyl‐p‐benzo quinone imine (NAPQI) modifies cysteine residues of KV7 subunits and reinforces channel gating in homomeric and heteromeric KV7.2 to KV7.5, but not in KV7.1 channels. In KV7.2, a triple cysteine motif located within the S2‐S3 linker region mediates this reinforcement that can be expected to reduce the excitability of nociceptors and to mediate antinociceptive actions of paracetamol. [ABSTRACT FROM AUTHOR]

Sea anemones are venomous animals that rely on their venom for prey capture, defense against predators, and intraspecific competition. Currently, comprehensive molecular and evolutionary analyses of the toxin repertoire for sea anemones are limited by a lack of proteomic data for most species. In this study, proteo-transcriptomic analysis was used to expand our knowledge of the proteinaceous components of sea anemone venom by determining the secreted venom proteome of Calliactis polypus. Electromechanical stimulation was used to obtain the secreted venom of C. polypus. We identified a low complexity proteome that was dominated by toxins with similarity to known neurotoxins, as well as six novel toxin candidates. The novel putative toxin candidates were found to be taxonomically restricted to species from the superfamily Metridioidea. Furthermore, the secreted venom of C. polypus had only three putative toxins in common with the venom of acontia from the same species and little similarity with the secreted venom of closely related species. Overall, this demonstrates that regionalized and lineage-specific variability in toxin abundance is common among sea anemone species. Moreover, the limited complexity of the toxin repertoire found in C. polypus supports the idea that peptide neurotoxins make up the dominant toxin arsenal found in the venom of sea anemones. [ABSTRACT FROM AUTHOR]

Venom systems are complex traits that have independently emerged multiple times in diverse plant and animal phyla. Within each venomous lineage there typically exists interspecific variation in venom composition where several factors have been proposed as drivers of variation, including phylogeny and diet. Understanding these factors is of broad biological interest and has implications for the development of antivenom therapies and venom-based drug discovery. Because of their high species richness and the presence of several major evolutionary prey shifts, venomous marine cone snails (genus Conus) provide an ideal system to investigate drivers of interspecific venom variation. Here, by analyzing the venom gland expression profiles of ∼3,000 toxin genes from 42 species of cone snail, we elucidate the role of prey-specific selection pressures in shaping venom variation. By analyzing overall venom composition and individual toxin structures, we demonstrate that the shifts from vermivory to piscivory in Conus are complemented by distinct changes in venom composition independent of phylogeny. In vivo injections of venom from piscivorous cone snails in fish further showed a higher potency compared with venom of nonpiscivores demonstrating a selective advantage. Together, our findings provide compelling evidence for the role of prey shifts in directing the venom composition of cone snails and expand our understanding of the mechanisms of venom variation and diversification. [ABSTRACT FROM AUTHOR]

Wrinkles, one of the most common signs of aging, are primarily caused by the continuous contraction of muscles. Muscle contraction is induced by the binding of acetylcholine (ACh), released at the neuromuscular junction, to nicotinic acetylcholine receptor (nAChR) present on the muscle cell surface. In this study, we aimed to develop a wrinkle-improving peptide that inhibits the binding of ACh to nAChR using peptide phage display technology. Our peptide showed a remarkably high binding affinity to nAChR subunit α1, with a value below 1 µM, and was found to inhibit the action of ACh through its interaction with these receptors. Furthermore, it increased collagen synthesis in skin cells and upregulated the expression of the aquaporin-3 (AQP3) and hyaluronan synthase-2 (HAS2) genes. These results confirm that the peptide effectively inhibits muscle contraction and enhances skin elasticity and hydration, contributing to its wrinkle-reducing effects. Clinical studies on humans observed significant improvement in wrinkles after three weeks of use, with substantial reduction observed after six weeks. In conclusion, these findings demonstrate the efficacy of the peptide (named Medipep) in reducing wrinkles. [ABSTRACT FROM AUTHOR]

As ecosystem disruptors and intermediate hosts for various parasites, freshwater snails have significant socioeconomic impacts on human health, livestock production, and aquaculture. Although traditional molluscicides have been widely used to mitigate these effects, their environmental impact has encouraged research into alternative, biologically based strategies to create safer, more effective molluscicides and diminish the susceptibility of snails to parasites. This review focuses on alterations in glucose metabolism in snails under the multifaceted stressors of parasitic infections, drug exposure, and environmental changes and proposes a novel approach for snail management. Key enzymes within the glycolytic pathway, such as hexokinase and pyruvate kinase; tricarboxylic acid (TCA) cycle; and electron transport chains, such as succinate dehydrogenase and cytochrome c oxidase, are innovative targets for molluscicide development. These targets can affect both snails and parasites and provide an important direction for parasitic disease prevention research. For the first time, this review summarises the reverse TCA cycle and alternative oxidase pathway, which are unique metabolic bypasses in invertebrates that have emerged as suitable targets for the formulation of low-toxicity molluscicides. Additionally, it highlights the importance of other metabolic pathways, including lactate, alanine, glycogenolysis, and pentose phosphate pathways, in snail energy supply, antioxidant stress responses, and drug evasion mechanisms. By analysing the alterations in key metabolic enzymes and their products in stressed snails, this review deepens our understanding of glucose metabolic alterations in snails and provides valuable insights for identifying new pharmacological targets. [ABSTRACT FROM AUTHOR]

The voltage-gated sodium (NaV) channel subtype NaV1.7 plays a critical role in pain signaling, making it an important drug target. Here we studied the molecular interactions between μ-Conotoxin KIIIA (KIIIA) and the human NaV1.7 channel (hNaV1.7). We developed a structural model of hNaV1.7 using Rosetta computational modeling and performed in silico docking of KIIIA using RosettaDock to predict residues forming specific pairwise contacts between KIIIA and hNaV1.7. We experimentally validated these contacts using mutant cycle analysis. Comparison between our KIIIA-hNaV1.7 model and the cryo-EM structure of KIIIA-hNaV1.2 revealed key similarities and differences between NaV channel subtypes with potential implications for the molecular mechanism of toxin block. The accuracy of our integrative approach, combining structural data with computational modeling, experimental validation, and molecular dynamics simulations, suggests that Rosetta structural predictions will be useful for rational design of novel biologics targeting specific NaV channels.

Methaqualone, a quinazolinone marketed commercially as Quaalude, is a central nervous system depressant that was used clinically as a sedative-hypnotic, then became a notorious recreational drug in the 1960s-80s. Due to its high abuse potential, medical use of methaqualone was eventually prohibited, yet it persists as a globally abused substance. Methaqualone principally targets GABAA receptors, which are the major inhibitory neurotransmitter-gated ion channels in the brain. The restricted status and limited accessibility of methaqualone have contributed to its pharmacology being understudied. Here, we use cryo-EM to localize the GABAA receptor binding sites of methaqualone and its more potent derivative, PPTQ, to the same intersubunit transmembrane sites targeted by the general anesthetics propofol and etomidate. Both methaqualone and PPTQ insert more deeply into subunit interfaces than the previously-characterized modulators. Binding of quinazolinones to this site results in widening of the extracellular half of the ion-conducting pore, following a trend among positive allosteric modulators in destabilizing the hydrophobic activation gate in the pore as a mechanism for receptor potentiation. These insights shed light on the underexplored pharmacology of quinazolinones and further elucidate the molecular mechanisms of allosteric GABAA receptor modulation through transmembrane binding sites. Methaqualone (Quaalude) is an infamous sedative-hypnotic, used in the past for treatment of insomnia. Here, the authors use structural biology and electrophysiology to elucidate its mechanism of action at its primary target, the GABAA receptor. [ABSTRACT FROM AUTHOR]

The vast biological and biochemical diversity of the global ocean is the driver behind marine bioprospecting for novel bioproducts. As Marine Biotechnology is gaining momentum as one of the main pillars of the ‘Brue Growth’ revolution, the ability to screen for novel compounds of interest in species with little or no genomic resources is paramount. With this respect, proteins, which are easily metabolised, can be synthetised using convenient DNA recombinant methods and can easily be modified to better meet the needs of human society, making them prized targets. Evidently, proteins that hold natural bioactivity and specificity such as toxins and other venom components, have long captured the focus of biotechnologists, leading to the merger between environmental omics and toxinology termed as ‘venomics’. Indeed, bioactive proteins such as conopeptides, conotoxins, turripeptides and others are long deemed important subjects of research. Even though current mainstream paradigms set the focus on secondary metabolites from marine organisms, transcriptomics and proteomics approaches and their combination are rising strategies for screening for thousands of proteins and peptides in non-conventional biological models, emphasising, but not limited to, marine invertebrate animals due to their abundance, biodiversity and uncanny biochemical strategies to cope with selective pressure in literally every known marine habitat. Untargeted approaches, such as RNA-Seq – based transcriptomics and tandem mass spectrometry – based proteomics, can circumvent limitations related with absent or reduced genomic annotation. The present review will outline the main contributions of ‘omics’ and computational approaches for bioprospecting for proteinaceous marine bioactives. Despite the relatively low number of ‘omics’ studies with the main purpose of discover novel compounds, there is already important literature showcasing pipelines and approaches for revolutionising the exploration of the ocean. [ABSTRACT FROM AUTHOR]

The escalating resistance of agricultural pests to chemical insecticides necessitates the development of novel, efficient, and safe biological insecticides. Conus quercinus, a vermivorous cone snail, yields a crude venom rich in peptides for marine worm predation. This study screened six α-conotoxins with insecticidal potential from a previously constructed transcriptome database of C. quercinus, characterized by two disulfide bonds. These conotoxins were derived via solid-phase peptide synthesis (SPPS) and folded using two-step iodine oxidation for further insecticidal activity validation, such as CCK-8 assay and insect bioassay. The final results confirmed the insecticidal activities of the six α-conotoxins, with Qc1.15 and Qc1.18 exhibiting high insecticidal activity. In addition, structural analysis via homology modeling and functional insights from molecular docking offer a preliminary look into their potential insecticidal mechanisms. In summary, this study provides essential references and foundations for developing novel insecticides. [ABSTRACT FROM AUTHOR]

In this research, the MIL-47/ACET/Nafion/GCE electrochemical senser for the determination of diethylstilbestrol (DES) was prepared with vanadyl sulfate (VOSO4· n H2O) and terephthalic acid (H2BDC) as the main raw materials, compounded with acetylene black (ACET) and perfluorosulfonic acid polymer (Nafion). The compound DES belongs to the category of estrogens, and prolonged exposure to the environment can have detrimental effects on the physiological functioning of both humans and animals. Due to the strong DES enrichment performance of MIL-47(V-MOFs) with large specific surface area, in addition to the excellent conductivity and electrocatalysis of composite materials, this modified senser had good electrochemical response to DES. With differential pulse voltammetry, in optimum condition of 0.1 M NaH2PO4–Na2HPO4 at pH = 7.0, potential interval of −1.0 to 1.0 V, enrichment time of 120 s and enrichment potential of 0.2 V, there was a good linear relationship between peak current and the concentration of DES over the range of 0.1 and 50 μ M, and the limit of detection was 0.008 μ M. The sensor accurately detected DES in actual water samples, with recovery rates ranging from 89.21% to 105.3%. The electrochemical sensor was simple to prepare and had practical significance for the detection of DES in water. The research results of the sensor provide another alternative analytical means for the sensitive detection of DES in the environment, which is important for maintaining public health. [ABSTRACT FROM AUTHOR]

A relaxin‐like gonad‐stimulating peptide (RGP), Aso‐RGP, featuring six cysteine residues, was identified in the Crown‐of‐Thorns Starfish (COTS, Acanthaster cf. solaris) and initially produced through recombinant yeast expression. This method yielded a single‐chain peptide with an uncleaved C‐peptide (His Tag) and suboptimal purity. Our objective was to chemically synthesize Aso‐RGP in its mature form, comprising two chains (A and B) and three disulfide bridges, omitting the C‐peptide. Furthermore, we aimed to synthesize a newly identified relaxin‐like peptide, Aso‐RLP2, from COTS, which had not been previously synthesized. This paper reports the first total chemical synthesis of Aso‐RGP and Aso‐RLP2. Aso‐RGP synthesis proceeded without major issues, whereas the A‐chain of Aso‐RLP2, in its reduced and unfolded state with two free thiols, presented considerable challenges. These were initially marked by "messy" RP‐HPLC profiles, typically indicative of synthesis failure. Surprisingly, oxidizing the A‐chain significantly improved the RP‐HPLC profile, revealing the main issue was not synthesis failure but the peptide's aggregation tendency, which initially obscured analysis. This discovery highlights the critical need to account for aggregation in peptide synthesis and analysis. Ultimately, our efforts led to the successful synthesis of both peptides with purities exceeding 95 %. [ABSTRACT FROM AUTHOR]

Developing synthetic molecular devices for controlling ion transmembrane transport is a promising research field in supramolecular chemistry. These artificial ion channels provide models to study ion channel diseases and have huge potential for therapeutic applications. Compared with self‐assembled ion channels constructed by intermolecular weak interactions between smaller molecules or cyclic compounds, metallacage‐based ion channels have well‐defined structures and can exist as single components in the phospholipid bilayer. A naphthalene diimide‐based artificial chloride ion channel is constructed through efficient subcomponent self‐assembly and its selective ion transport activity in large unilamellar vesicles and the planar lipid bilayer membrane by fluorescence and ion‐current measurements is investigated. Molecular dynamics simulations and density functional theory calculations show that the metallacage spans the entire phospholipid bilayer as an unimolecular ion transport channel. This channel transports chloride ions across the cell membrane, which disturbs the ion balance of cancer cells and inhibits the growth of cancer cells at low concentrations. [ABSTRACT FROM AUTHOR]

Rattusin, an α-defensin-related antimicrobial peptide isolated from the small intestine of rats, has been previously characterized through NMR spectroscopy to elucidate its three-dimensional structure, revealing a C2 homodimeric scaffold stabilized by five disulfide bonds. This study aimed to identify the functional region of rattusin by designing and synthesizing various short analogs, subsequently leading to the development of novel peptide-based antibiotics. The analogs, designated as F1, F2, F3, and F4, were constructed based on the three-dimensional configuration of rattusin, among which F2 is the shortest peptide and exhibited superior antimicrobial efficacy compared to the wild-type peptide. The central cysteine residue of F2 prompted an investigation into its potential to form a dimer at neutral pH, which is critical for its antimicrobial function. This activity was abolished upon the substitution of the cysteine residue with serine, indicating the necessity of dimerization for antimicrobial action. Further, we synthesized β-hairpin-like analogs, both parallel and antiparallel, based on the dimeric structure of F2, which maintained comparable antimicrobial potency. In contrast to rattusin, which acts by disrupting bacterial membranes, the F2 dimer binds directly to DNA, as evidenced by fluorescence assays and DNA retardation experiments. Importantly, F2 exhibited negligible cytotoxicity up to 515 μg/mL, assessed via hemolysis and MTT assays, underscoring its potential as a lead compound for novel peptide-based antibiotic development. [ABSTRACT FROM AUTHOR]

Bacteria contain conserved mechanisms to control the intracellular levels of metal ions. Metalloregulatory transcription factors bind metal cations and play a central role in regulating gene expression of metal transporters. Often, these transcription factors regulate transcription by binding to a specific DNA sequence in the promoter region of target genes. Understanding the preferred DNA‐binding sequence for transcriptional regulators can help uncover novel gene targets and provide insight into the biological role of the transcription factor in the host organism. Here, we identify consensus DNA‐binding sequences and subsequent transcription regulatory networks for two metalloregulators from the ferric uptake regulator (FUR) and diphtheria toxin repressor (DtxR) superfamilies in Thermus thermophilus HB8. By homology search, we classify the DtxR homolog as a manganese‐specific, MntR (TtMntR), and the FUR homolog as a peroxide‐sensing, PerR (TtPerR). Both transcription factors repress separate ZIP transporter genes in vivo, and TtPerR acts as a bifunctional transcription regulator by activating the expression of ferric and hemin transport systems. We show TtPerR and TtMntR bind DNA in the presence of manganese in vitro and in vivo; however, TtPerR is unable to bind DNA in the presence of iron, likely due to iron‐mediated histidine oxidation. Unlike canonical PerR homologs, TtPerR does not appear to contribute to peroxide detoxification. Instead, the TtPerR regulon and DNA binding sequence are more reminiscent of Fur or Mur homologs. Collectively, these results highlight the similarities and differences between two metalloregulatory superfamilies and underscore the interplay of manganese and iron in transcription factor regulation. [ABSTRACT FROM AUTHOR]

The intricate relationships between parasites and hosts encompass a wide range of levels, from molecular interactions to population dynamics. Parasites influence not only the physiological processes in the host organism, but also the entire ecosystem, affecting mortality of individuals, the number of offspring through parasitic castration, and matter and energy cycles. Understanding the molecular mechanisms that govern host-parasite relationships and their impact on host physiology and environment remains challenging. In this study, we analyzed how infection with Microphallus trematodes affects the metabolome of two Littorina snail species inhabiting different intertidal zone shore levels. We applied non-targeted GC–MS-based metabolomics to analyze biochemical shifts induced by trematode infection in a host organism. We have identified changes in energy, amino acid, sugar, and lipid metabolism. In particular, we observed intensified amino acid catabolism and nitrogenous catabolites (glutamine, urea) production. These changes primarily correlated with infection and interspecies differences of the hosts rather than shore level. The changes detected in the host metabolism indicate that other aspects of life may have been affected, both within the host organism and at a supra-organismal level. Therefore, we explored changes in microbiota composition, deviations in the host molluscs behavior, and acetylcholinesterase activity (ACE, an enzyme involved in neuromuscular transmission) in relation to infection. Infected snails displayed changes in their microbiome composition. Decreased ACE activity in snails was associated with reduced mobility, but whether it is associated with trematode infection remains unclear. The authors suggest a connection between the identified biochemical changes and the deformation of the shell of molluscs, changes in their behavior, and the associated microbiome. The role of parasitic systems formed by microphallid trematodes and Littorina snails in the nitrogen cycle at the ecosystem level is also assumed. [ABSTRACT FROM AUTHOR]

Background: The potential molluscicidal extracts, obtained from indigenous plants Cannabis sativa, Acacia nilotica, and Tinospora cordifolia, were tested for toxicity against freshwater pulmonate snail Lymnaea acuminata, an intermediate host of Fasciola hepatica. The organic extracts had a significant effect on young snails. Materials and Methods: All organic extracts and column-purified fractions gave median lethal concentrations (19–100.05 mg/L; 24 h) that fell well within the threshold level of 100 mg/L, set for a potential molluscicide by the World Health Organization. Results: The toxicity of T. cordifolia stem acetone extract (96 h LC50: 16.08 mg/L) was more pronounced compared with C. sativa leaf ethanol extract (96 h LC50: 16.32 mg/L) and A. nilotica leaf ethanol extract (96 h LC50: 24.78 mg/L). β-caryophyllene, gallic acid, and berberine were characterized and identified as active molluscicidal components. Co-migration of β-caryophyllene (retardation factor [Rf] 0.95), gallic acid (Rf 0.30), and berberine (Rf 0.23) with column-purified parts of Cannabis sativa, Acacia nilotica, and Tinospora cordifolia on thin-layer chromatography demonstrates same Rf value, that is, 0.95, 0.30, and 0.23, respectively. Conclusion: This study indicates that these extracts thus represent potential plant-derived molluscicides that are worthy of further investigations. [ABSTRACT FROM AUTHOR]

The giant triton snail, Charonia tritonis (Linnaeus, 1758), crucial for coral reef ecosystems as a primary predator of the crown-of-thorns sea star, is experiencing a significant decline due to overfishing for its ornamental shell, underscoring the urgent need for conservation and deeper understanding of its role within marine biodiversity. This study presents the first complete mitogenome sequence of C. tritonis. Spanning 15,346 bp, the C. tritonis mitogenome comprises 13 protein-coding genes (PCGs), 22 tRNA genes, and two rRNA genes. Phylogenetic analysis of 88 Littorinimorpha mitogenomes confirms C. tritonis and C. lampas are grouped together within the family Charoniidae as a sister group to the remaining Tonnoidea families. This research not only enhances the taxonomic classification and conservation efforts for marine gastropods but also serves as a vital reference for future evolutionary and genetic studies within the Caenogastropoda. [ABSTRACT FROM AUTHOR]

Fasciola hepatica has a complex lifecycle with multiple intermediate and definitive hosts and influenced by environmental factors. The disease causes significant morbidity in children and its prevalent worldwide. There is lack of data about distribution and burden of the disease in endemic regions, owing to poor efficacy of the different diagnostic methods used. A novel PCR-based test was developed by using a portable mini-PCR® platform to detect Fasciola sp. DNA and interpret the results via a fluorescence viewer and smartphone image analyzer application. Human stool, snail tissue, and water samples were used to extract DNA. Primers targeting the ITS-1 of the 18S rDNA gene of Fasciola sp. were used. The limit of detection of the mini-PCR test was 1 fg/μL for DNA samples diluted in water, 10 fg/μL for Fasciola/snail DNA scramble, and 100 fg/μL for Fasciola/stool DNA scramble. The product detection by agarose gel, direct visualization, and image analyses showed the same sensitivity. The Fh mini-PCR had a sensitivity and specificity equivalent to real-time PCR using the same specimens. Testing was also done on infected human stool and snail tissue successfully. These experiments demonstrated that Fh mini-PCR is as sensitive and specific as real time PCR but without the use of expensive equipment and laboratory facilities. Further testing of multiple specimens with natural infection will provide evidence for feasibility of deployment to resource constrained laboratories. [ABSTRACT FROM AUTHOR]

The human voltage-gated sodium channel Nav1.7 is a widely proven target for analgesic drug studies. ProTx2, a 30-residue polypeptide from Peruvian green tarantula venom, shows high specificity to activity against human Nav1.7, suggesting its potential to become a non-addictive analgesic. However, its high sensitivity to human Nav1.4 raises concerns about muscle side effects. Here, we engineered three mutants (R13A, R13D, and K27Y) of ProTx2 to evaluate their pharmacological activities toward Nav1.7 and Nav1.4. It is demonstrated that the mutant R13D maintained the analgesic effect in mice while dramatically reducing its muscle toxicity compared with ProTx2. The main reason is the formation of a strong electrostatic interaction between R13D and the negatively charged amino acid residues in DII/S3-S4 of Nav1.7, which is absent in Nav1.4. This study advances our understanding and insights on peptide toxins, paving the way for safer, effective non-addictive analgesic development. [ABSTRACT FROM AUTHOR]

NaV1.4 is a voltage-gated sodium channel subtype that is predominantly expressed in skeletal muscle cells. It is essential for producing action potentials and stimulating muscle contraction, and mutations in NaV1.4 can cause various muscle disorders. The discovery of the cryo-EM structure of NaV1.4 in complex with β1 has opened new possibilities for designing drugs and toxins that target NaV1.4. In this review, we summarize the current understanding of channelopathies, the binding sites and functions of chemicals including medicine and toxins that interact with NaV1.4. These substances could be considered novel candidate compounds or tools to develop more potent and selective drugs targeting NaV1.4. Therefore, studying NaV1.4 pharmacology is both theoretically and practically meaningful. [ABSTRACT FROM AUTHOR]

[Display omitted] • Wider distribution of invasive snails compared with native snails in the Limpopo River system. • cox 1 genetic data shows multiple lineages of invasive snails outside their native ranges. • More digenean species occurred in Pseudosuccinea columella than in the other snails. • The present xiphidiocercaria represents an unknown species of Orientocreadium. Invasive snails are associated with ecological problems in freshwater bodies worldwide. However, their impact on the transmission of digenean infections remain underreported. In the present study, 1708 specimens representing four snail species were sampled from four impoundments in the Limpopo River system in South Africa. Gyraulus chinensis (Planorbidae), Physella acuta (Physidae) and Pseudosuccinea columella (Lymnaeidae), which are invasive, were found in all the sampling sites. In contrast, the native lymnaeid Radix natalensis occurred at only one study site. Digeneans were observed only from R. natalensis (prevalence = 49%) and Ps. columella (prevalence = 23%). Morphological and genetic analyses revealed four digeneans: Fasciola nyanzae , Orientocreadium sp., Petasiger sp. and Patagifer vioscai. Pseudosuccinea columella was infected by the four digeneans while R. natalensis harboured only Orientocreadium sp. and Petasiger sp. Partial sequences of Orientocreadium sp. from the current study differed from congeners whose DNA data are available on GenBank, by p-distances of at least 1.84 and 2.2% for 28S and the internal transcribed spacer (ITS) rDNA, respectively. Phylogenetic analyses demonstrated that the present species is sister to Orientocreadium batrachoides. Genetic and phylogenetic data based on 28S and ITS rDNA suggested that Petasiger sp. from the present study and isolates of three unidentified Petasiger spp. from Kenya, Hungary and Australia, were representatives of the same species. This is the first known report of Orientocreadium , Petasiger and Patagifer from Ps. columella. The occurrence of F. nyanzae in Ps. columella indicates spillback from R. natalensis. These findings echo the concerns raised in previous studies about the potential role of Ps. columella in the amplification of digenean diseases in its introduced range. Phylogenetic analyses of partial sequences of the cytochrome c oxidase subunit 1 mitochondrial gene (cox 1) showed multiple lineages of Ps. columella in North and South America. Pseudosuccinea columella specimens from the present study belong to an invasive genotype that has spread globally and has been reported from Zimbabwe, Egypt, Portugal, Australia, Argentina, Colombia and New Mexico (USA). Physella acuta from the current study had a stronger genetic relationship with isolates from Canada and Iceland, than with isolates from other parts of Africa, suggesting several invasion routes into Africa. This is the first known DNA characterisation of G. chinensis from Africa. Phylogenetic reconstruction indicated multiple exit events of G. chinensis from Asia into Europe and Africa. South African isolates clustered in a recent branch containing isolates from the Czech Republic and Hong Kong, China. Considering the presence of invasive snails in all the sampling sites in the present study, it is necessary to investigate the factors that enhance their establishment and to monitor their effects on the native snail populations. [ABSTRACT FROM AUTHOR]

Cataract disease is strongly associated with progressively accumulating oxidative damage to the extremely long-lived crystallin proteins of the lens. Cysteine oxidation affects crystallin folding, interactions, and light-scattering aggregation especially strongly due to the formation of disulfide bridges. Minimizing crystallin aggregation is crucial for lifelong lens transparency, so one might expect the ubiquitous lens crystallin superfamilies (α and βγ) to contain little cysteine. Yet, the Cys content of γ-crystallins is well above the average for human proteins. We review literature relevant to this longstanding puzzle and take advantage of expanding genomic databases and improved machine learning tools for protein structure prediction to investigate it further. We observe remarkably low Cys conservation in the βγ-crystallin superfamily; however, in γ-crystallin, the spatial positioning of Cys residues is clearly fine-tuned by evolution. We propose that the requirements of long-term lens transparency and high lens optical power impose competing evolutionary pressures on lens βγ-crystallins, leading to distinct adaptations: high Cys content in γ-crystallins but low in βB-crystallins. Aquatic species need more powerful lenses than terrestrial ones, which explains the high methionine content of many fish γ- (and even β-) crystallins. Finally, we discuss synergies between sulfur-containing and aromatic residues in crystallins and suggest future experimental directions. [ABSTRACT FROM AUTHOR]

Transposable elements (TEs) are repetitive DNA that can create genome structure and regulation variability. The genome of Rhizophagus irregularis , a widely studied arbuscular mycorrhizal fungus (AMF), comprises ∼50% repetitive sequences that include TEs. Despite their abundance, two-thirds of TEs remain unclassified, and their regulation among AMF life stages remains unknown. Here, we aimed to improve our understanding of TE diversity and regulation in this model species by curating repeat datasets obtained from chromosome-level assemblies and by investigating their expression across multiple conditions. Our analyses uncovered new TE superfamilies and families in this model symbiont and revealed significant differences in how these sequences evolve both within and between R. irregularis strains. With this curated TE annotation, we also found that the number of upregulated TE families in colonized roots is 4 times higher than in the extraradical mycelium, and their overall expression differs depending on the plant host. This work provides a fine-scale view of TE diversity and evolution in model plant symbionts and highlights their transcriptional dynamism and specificity during host–microbe interactions. We also provide Hidden Markov Model profiles of TE domains for future manual curation of uncharacterized sequences (https://github.com/jordana-olive/TE-manual-curation/tree/main). [ABSTRACT FROM AUTHOR]

Aim: We aimed to understand how biogeographical processes and moisture niche ecology contributed to the spatio‐temporal diversification dynamics in the land snail genus Vertigo. Location: Global (North America, Europe, Asia, Africa). Time Period: Cenozoic era. Major Taxa Studied: Minute terrestrial snails of the genus Vertigo. Methods: We reconstructed a time‐calibrated phylogeny of 94 Vertigo taxa (~85% of all known extant species) based on mitochondrial and nuclear DNA data. Leveraging this phylogeny with distributional and ecological data from >7000 populations, we performed biogeographic and ecological modelling to investigate evolutionary mechanisms of global Vertigo diversification. Results: Vertigo has diversified since the Early Eocene, ca. 47.6 Ma (95% HPD = 46.0–52.7), with its six subgenera originating from the Late Eocene (30.2–48.7 Ma) to the Early Miocene (13.3–23.0 Ma). Species diversity accumulated linearly, with a slight increase 35–30 and 25–20 Ma, coinciding with the emergence of most subgenera and northern hemisphere cooling, respectively. Biogeographic modelling indicated that most diversification events occurred in sympatry (no range modification), but that rare founder events drove global diversification. Soil moisture conditions, a major variable defining Vertigo niches, displayed significant phylogenetic signal, but varied less among subgenera relative to within. Shifts in biogeographical ranges and moisture niches (or the absence thereof) were significantly associated at macroevolutionary scales, with most niche shifts upon sympatric cladogenesis and hardly any upon founder‐event speciation. Main Conclusions: Our results indicate that ecological shifts in soil moisture niches occasionally drove cladogenesis in sympatry and anagenetic range extensions, but that long‐distance dispersal was mainly successful in the absence of such shifts. A combination of neutral (founder events and drift) and selective mechanisms (adaptive habitat shifts) has determined the macroevolutionary success of Vertigo. Our results imply that species with high niche fidelity or fewer opportunities for long‐distance dispersal will be more vulnerable to future anthropogenic stressors. [ABSTRACT FROM AUTHOR]

Pathogenic variants in the Crumbs homolog 1 (CRB1) gene lead to severe, childhood-onset retinal degeneration leading to blindness in early adulthood. There are no approved therapies, and traditional adeno-associated viral vector-based gene therapy approaches are challenged by the existence of multiple CRB1 isoforms. Here, we describe three CRB1 variants, including a novel, previously unreported variant that led to retinal degeneration. We offer a CRISPR-Cas-mediated DNA base editing strategy as a potential future therapeutic approach. This study is a retrospective case series. Clinical and genetic assessments were performed, including deep phenotyping by retinal imaging. In silico analyses were used to predict the pathogenicity of the novel variant and to determine whether the variants are amenable to DNA base editing strategies. Case 1 was a 24-year-old male with cone–rod dystrophy and retinal thickening typical of CRB1 retinopathy. He had a relatively preserved central outer retinal structure and a best corrected visual acuity (BCVA) of 60 ETDRS letters in both eyes. Genetic testing revealed compound heterozygous variants in exon 9: c.2843G>A, p.(Cys948Tyr) and a novel variant, c.2833G>A, p.(Gly945Arg), which was predicted to likely be pathogenic by an in silico analysis. Cases 2 and 3 were two brothers, aged 20 and 24, who presented with severe cone–rod dystrophy and a significant disruption of the outer nuclear layers. The BCVA was reduced to hand movements in both eyes in Case 2 and to 42 ETDRS letters in both eyes in Case 3. Case 2 was also affected with marked cystoid macular lesions, which are common in CRB1 retinopathy, but responded well to treatment with oral acetazolamide. Genetic testing revealed two c.2234C>T, p.(Thr745Met) variants in both brothers. As G-to-A and C-to-T variants, all three variants are amenable to adenine base editors (ABEs) targeting the forward strand in the Case 1 variants and the reverse strand in Cases 2 and 3. Available PAM sites were detected for KKH-nSaCas9-ABE8e for the c.2843G>A variant, nSaCas9-ABE8e and KKH-nSaCas9-ABE8e for the c.2833G>A variant, and nSpCas9-ABE8e for the c.2234C>T variant. In this case series, we report three pathogenic CRB1 variants, including a novel c.2833G>A variant associated with early-onset cone–rod dystrophy. We highlight the severity and rapid progression of the disease and offer ABEs as a potential future therapeutic approach for this devastating blinding condition. [ABSTRACT FROM AUTHOR]

This study uses the optimal (custom) design to focus on the supermasscolloider wet milling process for synthesizing calcium carbonate nanoparticles from the Achatina Fulica using the optimal (custom) design. The snail shell particle size of 150 μm from dry milling was wet milled to get smaller particle sizes. The response surface design method was used to investigate the effect of independent parameters such as consistency (amount of microparticle loading in calcium carbonate suspension) and the number of runs on the particle size of calcium carbonate produced using a supermasscolloider. Results show that the microparticle loading in calcium carbonate suspension and the number of runs significantly affected the synthesized particles. The increase in microparticle loading reduced the effectiveness of the supermasscolloider blade, resulting in the production of particle sizes ranging from 21- to 40 nm. A higher number of runs with a smaller loading of microparticles in calcium carbonate suspension offered a fine particle size of 21.30 nm. The experimental data quartic polynomial models gave a coefficient of determination (R2) of 0.92. The optimum milling runs of 750 and 1% consistency microparticle loading produced a calcium carbonate nanoparticle size of 21 nm. This technique has shown that calcium carbonate nanoparticles can be produced at low cost, at less period, with low agglomeration from Achatina Fulica shell using supermasscolloider. [ABSTRACT FROM AUTHOR]

Nociceptive sensory neurons convey pain-related signals to the CNS using action potentials. Loss-of-function mutations in the voltage-gated sodium channel NaV1.7 cause insensitivity to pain (presumably by reducing nociceptor excitability) but clinical trials seeking to treat pain by inhibiting NaV1.7 pharmacologically have struggled. This may reflect the variable contribution of NaV1.7 to nociceptor excitability. Contrary to claims that NaV1.7 is necessary for nociceptors to initiate action potentials, we show that nociceptors can achieve similar excitability using different combinations of NaV1.3, NaV1.7, and NaV1.8. Selectively blocking one of those NaV subtypes reduces nociceptor excitability only if the other subtypes are weakly expressed. For example, excitability relies on NaV1.8 in acutely dissociated nociceptors but responsibility shifts to NaV1.7 and NaV1.3 by the fourth day in culture. A similar shift in NaV dependence occurs in vivo after inflammation, impacting ability of the NaV1.7-selective inhibitor PF-05089771 to reduce pain in behavioral tests. Flexible use of different NaV subtypes exemplifies degeneracy - achieving similar function using different components - and compromises reliable modulation of nociceptor excitability by subtype-selective inhibitors. Identifying the dominant NaV subtype to predict drug efficacy is not trivial. Degeneracy at the cellular level must be considered when choosing drug targets at the molecular level. [ABSTRACT FROM AUTHOR]

Background: Polymyxin B (PMB) and polymyxin E (colistin, CST) are polymyxin antibiotics, which are considered last-line therapeutic options against multidrug-resistant Gram-negative bacteria in serious infections. However, there is increasing risk of resistance to antimicrobial drugs. Effective efflux pump inhibitors (EPIs) should be developed to help combat efflux pump-mediated antibiotic resistance. Methods: Chryseobacterium sp. PL22-22A was isolated from aquaculture sewage under selection with 8  mg/L PMB, and then its genome was sequenced using Oxford Nanopore and BGISEQ-500 platforms. Cpr (Chryseobacterium Polymyxins Resistance) genes encoding a major facilitator superfamily-type tripartite efflux system, were found in the genome. These genes, and the gene encoding a truncation mutant of CprB from which sequence called CprBc was deleted, were amplified and expressed/co-expressed in Escherichia coli DH5α. Minimum inhibitory concentrations (MICs) of polymyxins toward the various E. coli heterologous expression strains were tested in the presence of 2–128  mg/L PMB or CST. The pumping activity of CprABC was assessed via structural modeling using Discovery Studio 2.0 software. Moreover, the influence on MICs of baicalin, a novel MFS EPI, was determined, and the effect was analyzed based on homology modeling. Results: Multidrug-resistant bacterial strain Chryseobacterium sp. PL22-22A was isolated in this work; it has notable resistance to polymyxin, with MICs for PMB and CST of 96 and 128  mg/L, respectively. A novel MFS-type tripartite efflux system, named CprABC, was identified in the genome of Chryseobacterium sp. PL22-22A. Heterologous expression and EPI assays indicated that the CprABC system is responsible for the polymyxin resistance of Chryseobacterium sp. PL22-22A. Structural modeling suggested that this efflux system provides a continuous conduit that runs from the CprB funnel through the CprC porin domain to pump polymyxins out of the cell. A specific C-terminal α-helix, CprBc, has an activation function on polymyxin excretion by CprB. The flavonoid compound baicalin was found to affect the allostery of CprB and/or obstruct the substrate conduit, and thus to inhibit extracellular polymyxin transport by CprABC. Conclusion: Novel MFS-type tripartite efflux system CprABC in Chryseobacterium sp. PL22-22A mediates resistance to polymyxins, and baicalin is a promising E [ABSTRACT FROM AUTHOR]

Scavenger receptors are a protein superfamily that typically consists of one or more repeats of the scavenger receptor cysteine-rich structural domain (SRCRD), which is an ancient and highly conserved protein module. The expression and purification of eukaryotic proteins containing multiple disulfide bonds has always been challenging. The expression systems that are commonly used to express SRCRD proteins mainly consist of eukaryotic protein expression systems. Herein, we established a high-level expression strategy of a Type B SRCRD unit from human salivary agglutinin using the Escherichia coli expression system, followed by a refolding and purification process. The untagged recombinant SRCRD was expressed in E. coli using the pET-32a vector, which was followed by a refolding process using the GSH/GSSG redox system. The SRCRD expressed in E. coli SHuffle T7 showed better solubility after refolding than that expressed in E. coli BL21(DE3), suggesting the importance of the disulfide bond content prior to refolding. The quality of the refolded protein was finally assessed using crystallization and crystal structure analysis. As proteins refolded from inclusion bodies exhibit a high crystal quality and reproducibility, this method is considered a reliable strategy for SRCRD protein expression and purification. To further confirm the structural integrity of the refolded SRCRD protein, the purified protein was subjected to crystallization using sitting-drop vapor diffusion method. The obtained crystals of SRCRD diffracted X-rays to a resolution of 1.47 Å. The solved crystal structure appeared to be highly conserved, with four disulfide bonds appropriately formed. The surface charge distribution of homologous SRCRD proteins indicates that the negatively charged region at the surface is associated with their calcium-dependent ligand recognition. These results suggest that a high-quality SRCRD protein expressed by E. coli SHuffle T7 can be successfully folded and purified, providing new options for the expression of members of the scavenger receptor superfamily. [ABSTRACT FROM AUTHOR]

Disulfides in peptides and proteins are essential for maintaining a properly folded structure. Their oxidative folding is invariably performed in an aqueous‐buffered solution. However, this process is often slow and can lead to misfolded products. Here, we report a novel concept and strategy that is bio‐inspired to mimic protein disulfide isomerase (PDI) by accelerating disulfide exchange rates many thousand‐fold. The proposed strategy termed organic oxidative folding is performed under organic solvents to yield correctly folded cysteine‐rich microproteins instantaneously without observable misfolded or dead‐end products. Compared to conventional aqueous oxidative folding strategies, enormously large rate accelerations up to 113,200‐fold were observed. The feasibility and generality of the organic oxidative folding strategy was successfully demonstrated on 15 cysteine‐rich microproteins of different hydrophobicity, lengths (14 to 58 residues), and numbers of disulfides (2 to 5 disulfides), producing the native products in a second and in high yield. [ABSTRACT FROM AUTHOR]

Schistosomiasis is a neglected tropical disease caused by parasitic worms of the genus Schistosoma. In Brazil, there are reports of infection by the Schistosoma mansoni species, which has the Biomphalaria glabrata snail as one of its intermediate hosts. The present work aimed to test the effects of different Abelmoschus esculentus seed extracts and fractions on adults and embryos of B. glabrata and S. mansoni cercariae. A total of four crude extracts and thirteen fractions with different organic solvents were used for the bioassays. The extracts were analyzed using high-performance liquid chromatography coupled to mass spectrometry. Molluscicidal activity was assessed in 24-well plates, after which the LC50 and LC90 were calculated. Assays with B. glabrata embryos and S. mansoni cercariae were also performed. These findings indicate the presence of flavanoids in the hexane, ethyl acetate, and ethanol crude extracts. For the molluscicidal activity assays, eight fractions had an LC90 value less than that recommended by the WHO. The methanol fraction of the dichloromethane extract (FrMeOH EDM) had the most promising results, with an LC90 of 37.15 mg/L and 100% mortality in embryos of B. glabrata and cercariae. FrMeOH, EDM, and other fractions are possible candidates for new drugs for the combat of schistosomiasis. [ABSTRACT FROM AUTHOR]

Peptide biomolecules have important bioactivities and hence their use in drug design and development studies has increased in recent years. Conotoxins are natural peptides that obtained from cone snail venoms and have the potential to be used for chronic pain treatment, Parkinson's disease, schizophrenia, obesity and cancer due to their effects on the nervous system. However, their use as medicines has been limited as they can be easily broken down by many proteolytic enzymes in the body. Several modification methods are used to overcome these disadvantages. Cyclization of the peptide backbone is one such method and has been used to stabilize various linear peptides. In this study, the N- and C-termini of a-conotoxin TxID with two disulfide bridges were joined using a six amino acid long GGAAGG linker peptide chain to cyclize the peptide backbone and the serum stability of the cyclized peptide was examined. The cyclic TxID peptide remained intact about 50% in human serum after 24 hours. [ABSTRACT FROM AUTHOR]

Cyclotides are plant-derived peptides characterized by a head-to-tail cyclic backbone and a cystine knot motif comprised of three disulfide bonds. Formation of this motif via in vitro oxidative folding can be challenging and can result in misfolded isomers with nonnative disulfide connectivities. Here, we investigated the effect of β-turn nucleation on cyclotide oxidative folding. Two types of β-turn mimics were grafted into kalata B1, individually replacing each of the four β- turns in the folded cyclotide. Insertion of D-Pro-Gly into loop 5 was beneficial to the folding of both cyclic kB1 and a linear form of the peptide. The linear grafted analog folded fourtimes faster in aqueous conditions than cyclic kB1 in optimized conditions. Additionally, the cyclic analogue folded without the need for redox agents by transitioning through a native-like intermediate that was on-pathway to product formation. Kalata B1 is from the Möbius subfamily of cyclotides. Grafting D-Pro-Gly into loop 5 of cyclotides from two other subfamilies also had a beneficial effect on folding. Our findings demonstrate the importance of a β-turn nucleation site for cyclotide oxidative folding, which could be adopted as a chemical strategy to improve the in vitro folding of diverse cystine-rich peptides. [ABSTRACT FROM AUTHOR]

Eulimidae is a highly diverse family of gastropods that are often parasites of echinoderms. They are cosmopolitan and live from the intertidal to great depths. Despite its wide geographic and bathymetric distribution, no species of Eulimidae have been reported for the Salas & Gómez Ridge to date. In this study, we describe Melanella martarum sp. nov., which was collected during the EPIC oceanographic cruise onboard RV Mirai (JAMSTEC, Japan) in 2019. Seven specimens were collected with a modified Agassiz trawl on the summit of seamount "Pearl'' (Zhemchuznaya) in the Salas & Gómez Ridge (25.59°S, 89.13_W) at 545 m depth. The morphology of M. martarum sp. nov. was compared with other Melanella species reported for the area, including Chile and Rapa Nui. DNA was extracted and partial sequences of the mitochondrial genes Cytochrome Oxidase 1 (COI) and 16S rDNA, and the nuclear gene Histone 3 (H3) were sequenced. Melanella martarum sp. nov. has morphological characteristics that separate it from other species of Melanella, such as the thickness and color of the shell, and the shape of the protoconch. In addition, M. martarum sp. nov. was genetically differentiated from other Melanella spp. sequences (uncorrected p distances from 18,1_8.6% in mitochondrial COI and 16S rDNA to 3% in nuclear H3 sequences). Although there is not much molecular data available for Eulimidae, the phylogenetic analysis confirms the results obtained by morphology, placing the species found on the Salas & Gómez Ridge within the genus Melanella. The current study advances the understanding of the poorly known benthic fauna found on seamounts in the easternmost part of the Sala & Gómez ridge, a location distinguished by a high level of endemism. [ABSTRACT FROM AUTHOR]

Conotoxins are small and highly potent neurotoxic peptides derived from the venom of marine cone snails which have captured the interest of the scientific community due to their pharmacological potential. These toxins display significant sequence and structure diversity, which results in a wide range of specificities for several different ion channels and receptors. Despite the recognized importance of these compounds, our ability to determine their binding targets and toxicities remains a significant challenge. Predicting the target receptors of conotoxins, based solely on their amino acid sequence, remains a challenge due to the intricate relationships between structure, function, target specificity, and the significant conformational heterogeneity observed in conotoxins with the same primary sequence. We have previously demonstrated that the inclusion of post-translational modifications, collisional cross sections values, and other structural features, when added to the standard primary sequence features, improves the prediction accuracy of conotoxins against non-toxic and other toxic peptides across varied datasets and several different commonly used machine learning classifiers. Here, we present the effects of these features on conotoxin class and molecular target predictions, in particular, predicting conotoxins that bind to nicotinic acetylcholine receptors (nAChRs). We also demonstrate the use of the Synthetic Minority Oversampling Technique (SMOTE)-Tomek in balancing the datasets while simultaneously making the different classes more distinct by reducing the number of ambiguous samples which nearly overlap between the classes. In predicting the alpha, mu, and omega conotoxin classes, the SMOTE-Tomek PCA PLR model, using the combination of the SS and P feature sets establishes the best performance with an overall accuracy (OA) of 95.95%, with an average accuracy (AA) of 93.04%, and an f1 score of 0.959. Using this model, we obtained sensitivities of 98.98%, 89.66%, and 90.48% when predicting alpha, mu, and omega conotoxin classes, respectively. Similarly, in predicting conotoxins that bind to nAChRs, the SMOTE-Tomek PCA SVM model, which used the collisional cross sections (CCSs) and the P feature sets, demonstrated the highest performance with 91.3% OA, 91.32% AA, and an f1 score of 0.9131. The sensitivity when predicting conotoxins that bind to nAChRs is 91.46% with a 91.18% sensitivity when predicting conotoxins that do not bind to nAChRs.

There is considerable interest in developing anti-glioma nanoplatforms. They make the all-in-one combination of therapies possible. Here we show how the selective Glioblastoma multiforme (GBM) cell killing of the here-established nanoplatforms increased after each coating and how the here-established vibration-inducing Alternating magnetic field (AMF) decreased the treatment time from 72 h to 30 s. Thanks to their magnetite core, these nanoplatforms can be guided to the tumor's specific site by a Fixed magnetic field, they bypass the Blood–Brain Barrier (BBB) and accumulate at the tumor site thanks to the RVG29 bonding to the G-protein on the ion-gated channel receptor known as the nicotinic acetylcholine receptor (nAchR), which expresses on BBB cells and overexpresses on GBM cells, and thanks to the positive charge gained by both chitosan and RVG29's peptide. Both ZIF-8 and its mediate adherence, Chitosan increases the drug loading capacity that stimuli response to the tumor's acidic environment. The Zn2+ ions generated from ZIF-8 sustained degradation in such an environment kill the GBM cells. Dynamic Light Scattering (DLS) evaluated these nanoplatform's mean size 155 nm indicating their almost optimum size for brain applications. Based on their elements' intrinsic properties, these nanoplatforms can enhance and combine other adjuvant therapies. [ABSTRACT FROM AUTHOR]

The monospecific genus Tartessiberus was described in the year 2021 including a single species (T. cilbanus). However, its description relied solely on morphological and anatomical data. In the present work, we use a fraction of the mitochondrial DNA cytochrome oxidase subunit I (COI), 16S ribosomal RNA (16S rRNA) and the nuclear large ribosomal subunit (LSU) to clarify its validity through phylogenetic positioning. Knowledge of the distribution of this species is also improved by citing new locations and expanding the geographical range to approximately 200 km2. Additionally, a morphometric analysis of 259 shells is presented for comparisons with shells of the Iberus marmoratus complex and testing the power of conchological features as a tool for specimen identification. The relatively high conchological variability found for T. cilbanus, together with the discovery of populations with intermediate conchological features between T. cilbanus and other closely related taxa, suggest that the determination of this species should be based on genetic criteria. Our molecular analyses demonstrate that T. cilbanus belongs to the Iberus genus, and thus, we proceed to update its taxonomic status to Iberus cilbanus comb. nov., and, thus, to consider Tartessiberus from now on as a junior synonym of Iberus. [ABSTRACT FROM AUTHOR]

Flupyradifurone (FLU) is a novel butenolide insecticide with partial agonist activity for insect nicotinic acetylcholine receptors. Its safety for non‐target organisms has been questioned in the literature, despite initial claims of its harmlessness. Detailed understanding of its toxicity and related molecular mechanisms remain under discussion. Thus, in this work, an optimized set of CHARMM compatible parameters for FLU is presented. CHARMM General Force Field program was used as a starting point while the non‐bonded and bonded parameters were adjusted and optimized to reproduce MP2/6‐31G(d) accuracy level results. For the validity assessment of these parameters, infrared spectrum, water‐octanol partition coefficient, and normal modes were computed and compared to experimental values found in the literature. Several MD simulations of FLU in water and FLU in complex with an acetylcholine‐binding protein were performed to estimate the ability of the optimized parameters to correctly describe its torsional space and reproduce observed crystallographic trends respectively. [ABSTRACT FROM AUTHOR]

The most promising anticonvulsant phytocompounds were explored in this work using docking, molecular dynamic (MD) simulation, and Molecular Mechanics-Poisson–Boltzmann Surface Area (MM-PBSA) approaches. A total of 70 phytochemicals were screened against α-amino-3-hydroxyl-5-methyl-4-isoxazole propionic acid (AMPA), N-methyl-d-aspartate (NMDA), voltage-gated sodium ion channels (VGSC), and carbonic anhydrase enzyme II (CA II) receptors, and the docking results were compared to the reference drug phenytoin. Amentoflavone displayed the highest affinity for AMPA and VGSC receptors, with docking scores of − 10.4 and − 10.1 kcal/mol, respectively. Oliganthin H-NMDA and epigallocatechin-3-gallate-CA II complexes showed docking scores of − 10.9 and − 6.9 kcal/mol, respectively. All four complexes depicted a high dock score compared to the phenytoin complex at the binding site of the corresponding proteins. The MD simulation investigated the stabilities and favorable conformation of apoproteins and ligand/reference-bound complexes. The results revealed that proteins AMPA, VGSC, and CA II were more efficiently stabilized by lead phytochemicals than phenytoin binding. Additionally, principal component analysis and MM-PBSA results suggested that these lead phytocompounds have good compactness and strong binding free energy. Further, physicochemical and pharmacokinetic studies revealed that these final lead phytochemicals would be suitable for oral intake, have sufficient intestinal permeability, and have the ability to cross the blood–brain barrier (BBB). Comprehensively, this study predicted amentoflavone as the best lead phytochemical out of the 70 anticonvulsant phytocompounds that can be used to treat epilepsy. [ABSTRACT FROM AUTHOR]

In accidents involving Crotalus snakes, the crotoxin complex (CTX) plays lethal action due to its neurotoxic activity. On the other hand, CTX have potential biotechnological application due to its anti-tumoral, anti-inflammatory, antimicrobial, analgesic and immunomodulatory properties. CTX is a heterodimer composed of Crotoxin A (CA or crotapotin), the acidic nontoxic and non-enzymatic component and; Crotoxin B (CB), a basic, toxic and catalytic PLA 2. Currently, there are two classes of CTX isoforms, whose differences in their biological activities have been attributed to features presented in CB isoforms. Here, we present the crystal structure of CB isolated from the Crotalus durissus collilineatus venom. It amino acid sequence was assigned using the SEQUENCE SLIDER software, which revealed that the crystal structure is a heterodimer composed of two new CB isoforms (col CB-A and col CB-B). Bioinformatic and biophysical analyses showed that the toxin forms a tetrameric assembly in solution similar to CB from Crotalus durissus terrificus venom, despite some differences observed at the dimeric interface. By the previously proposed classification, the col CB-B presents features of the class I isoforms while col CB-A cannot be classified into classes I and II based on its amino acid sequence. Due to similar features observed for other CB isoforms found in the NCBI database and the results obtained for col CB-A, we suggest that there are more than two classes of CTX and CB isoforms in crotalic venoms. • The crystal structure of CB from C. d. collilineatus venom was obtained. • Crystal structure is a heterodimer composed of two new CB isoforms. • Bioinformatic and biophysical analyses showed that the toxin is tetrameric. • col CB-A isoform cannot be classified into classes I and II based on its sequence. • We hypothesized that there are more than two classes of CB isoforms. [ABSTRACT FROM AUTHOR]

Conus marine snails can be identified by their venom compositions, which are complex and comprise a wide variety of pharmacologically potent peptides known as conotoxins. Due to their possible therapeutic usages, these peptides have attracted a lot of attention, especially in the field of oncology. This work aimed to study the cytotoxic effects of venom gland and venom tube extracts from Conus textile on three human cancer cell lines (namely HepG2, hepatocellular carcinoma, Caco2, colorectal cancer, Mcf7, and breast cancer). The present study assessed the venoms' half-maximal inhibitory concentrations (IC50), which represented their effectiveness in lowering cell viability, using an in-vitro experiment. With IC50 values ranging between 94 and 381.97µg/ml, the results showed that both venom extracts had strong cytotoxic effects on all examined cell lines. When compared to the gland extract, the venom tube extract consistently showed greater potency, which may indicate a higher concentration of active cytotoxic chemicals. When treated with venom tube extract, the Mcf7 cell line showed the lowest IC50 value, suggesting a promising potential for breast cancer therapies. These findings supported the hypothesis that Conus textile venom contains bioactive components with selective toxicity towards cancer cell lines. The insignificant standard deviations reported support the regularity and dependability of the cytotoxic effects. The present work provided the foundation for future purifying and mechanistic investigations of conotoxins and advanced the investigation of marine natural products as a source of potential anti-cancer drugs. [ABSTRACT FROM AUTHOR]

Viruses are one of the leading causes of human disease, and many highly pathogenic viruses still have no specific treatment drugs. Therefore, producing new antiviral drugs is an urgent matter. In our study, we first found that the natural wasp venom peptide Protopolybia-MP III had a significant inhibitory effect on herpes simplex virus type 1 (HSV-1) replication in vitro by using quantitative real-time PCR (qPCR), Western blotting, and plaque-forming assays. Immunofluorescence analysis showed Protopolybia-MP III could enter cells, and it inhibited multiple stages of the HSV-1 life cycle, including the attachment, entry/fusion, and post-entry stages. Furthermore, ultracentrifugation and electron microscopy detected that Protopolybia-MP III significantly suppressed HSV-1 virion infectivity at different temperatures by destroying the integrity of the HSV-1 virion. Finally, by comparing the antiviral activity of Protopolybia-MP III and its mutants, a series of peptides with better anti-HSV-1 activity were identified. Overall, this work found the function and mechanism of the antiviral wasp venom peptide Protopolybia-MP III and its derivatives against HSV-1 and laid the foundation for the research and development of wasp venom-derived antiviral candidate peptide drugs. [ABSTRACT FROM AUTHOR]

The neritid snail Theodoxus fluviatilis is found across habitats differing in salinity, from shallow waters along the coast of the Baltic Sea to lakes throughout Europe. Living close to the water surface makes this species vulnerable to changes in salinity in their natural habitat, and the lack of a free-swimming larval stage limits this species' dispersal. Together, these factors have resulted in a patchy distribution of quite isolated populations differing in their salinity tolerances. In preparation for investigating the mechanisms underlying the physiological differences in osmoregulation between populations that cannot be explained solely by phenotypic plasticity, we present here an annotated draft genome assembly for T. fluviatilis , generated using PacBio long reads, Illumina short reads, and transcriptomic data. While the total assembly size (1045 kb) is similar to those of related species, it remains highly fragmented (N scaffolds = 35,695; N50 = 74 kb) though moderately high in complete gene content (BUSCO single copy complete: 74.3%, duplicate: 2.6%, fragmented: 10.6%, missing: 12.5% using metazoa n = 954). Nevertheless, we were able to generate gene annotations of 21,220 protein-coding genes (BUSCO single copy complete: 65.1%, duplicate: 16.7%, fragmented: 9.1%, missing: 9.1% using metazoa n = 954). Not only will this genome facilitate comparative evolutionary studies across Gastropoda, as this is the first genome assembly for the basal snail family Neritidae, it will also greatly facilitate the study of salinity tolerance in this species. Additionally, we discuss the challenges of working with a species where high molecular weight DNA isolation is very difficult. [ABSTRACT FROM AUTHOR]

D‐peptide ligands can be screened for therapeutic potency and enzymatic stability using synthetic mirror‐image proteins (D‐proteins), but efficient acquisition of these D‐proteins can be hampered by the need to accomplish their in vitro folding, which often requires the formation of correctly linked disulfide bonds. Here, we report the finding that temporary installation of natural O‐linked‐β‐N‐acetyl‐D‐glucosamine (O‐GlcNAc) groups onto selected D‐serine or D‐threonine residues of the synthetic disulfide‐bonded D‐proteins can facilitate their folding in vitro, and that the natural glycosyl groups can be completely removed from the folded D‐proteins to afford the desired chirally inverted D‐protein targets using naturally occurring O‐GlcNAcase. This approach enabled the efficient chemical syntheses of several important but difficult‐to‐fold D‐proteins incorporating disulfide bonds including the mirror‐image tumor necrosis factor alpha (D‐TNFα) homotrimer and the mirror‐image receptor‐binding domain of the Omicron spike protein (D‐RBD). Our work establishes the use of O‐GlcNAc to facilitate D‐protein synthesis and folding and proves that D‐proteins bearing O‐GlcNAc can be good substrates for naturally occurring O‐GlcNAcase. [ABSTRACT FROM AUTHOR]

A new species of pomatiopsid freshwater snail, Fenouilia undata Chen & He, sp. nov., is described from Guangxi, China, based on morphological and molecular evidence. The new species can be distinguished from its congeners by the following combination of characters: shell with low, prosocline, rounded axial ribs and fine spiral striae, broader than high; aperture broader than shell height; radula with lateral teeth have only two or three faint, wavy ridges on inner side. A molecular analysis of partial mitochondrial COI and 16S DNA sequences supports the systematic position of the new taxon. [ABSTRACT FROM AUTHOR]

This paper reassesses the taxonomy and systematics of 11 arboreal snail species in the genus Amphidromus from Vietnam, Cambodia, Indonesia and Laos (A. bozhii Wang, 2019, A. buelowi Fruhstorfer, 1905, A. costifer Smith, 1893, A. haematostoma Möllendorff, 1898, A. ingens Möllendorff, 1900, A. madelineae Thach, 2020, A. metabletus Möllendorff, 1900, A. pankowskianus Thach, 2020, A. placostylus Möllendorff, 1900, A. roseolabiatus Fulton, 1896, and A. thachi Huber, 2015). The taxonomic validity of each species is supported by a phylogenetic analysis of mitochondrial COI and 16S rRNA gene fragments from 17 ingroup taxa. Amphidromus buelowi was found to comprise two populations from two distant localities, one from Mount Singgalang, West Sumatra, Indonesia and the other from southern Vietnam. The samples from southern Vietnam were previously described as A. asper Haas, 1934 and A. franzhuberi Thach, 2016, but they are now treated as junior synonyms of A. buelowi in this study. In addition, two species from Vietnam are described as new to science, viz. A. asperoides Jirapatrasilp & Lee, sp. nov. and A. ingensoides Jirapatrasilp & Lee, sp. nov., each of which is conchologically comparable to A. buelowi and A. ingens, respectively. [ABSTRACT FROM AUTHOR]