Structurally Diverse Alkaloids with Anti-Renal-Fibrosis Activity from the Centipede Scolopendra subspinipes mutilans.

Twelve new alkaloids, scolopenolines A-L (1-7, 9-11, 13, 14), along with six known analogues, were isolated from Scolopendra subspinipes mutilans, identified by analysis of spectroscopic data and quantum chemical and computational methods. Scolopenoline A (1), a unique guanidyl-containing C14 quinoline alkaloid, features a 6/6/5 ring backbone. Scolopenoline B (2) is a novel sulfonyl-containing heterodimer comprising quinoline and tyramine moieties. Scolopenoline G (7) presents a rare C12 quinoline skeleton with a 6/6/5 ring system. Alkaloids 1, 8, 10, and 15-18 display anti-inflammatory activity, while 10 and 16-18 also exhibit anti-renal-fibrosis activity. Drug affinity responsive target stability and RNA-interference assays show that Lamp2 might be a potentially important target protein of 16 for anti-renal-fibrosis activity.

Socialane, a Nonaprenyl Terpene Hydrocarbon Surface Lipid from the Collembola Hypogastrura socialis.

Springtails use unique compounds for their outermost epicuticular wax layer, often of terpenoid origin. We report here the structure and synthesis of socialane, the major cuticular constituent of the Collembola Hypogastrura socialis. Socialane is also the first regular nonaprenyl terpene with a cyclic head group. The saturated side chain has seven stereogenic centers, making the determination of the configuration difficult. We describe here the identification of socialane and a synthetic approach using the building blocks farnesol and phytol, enantioselective hydrogenation, and α-alkylation of sulfones for the synthesis of various stereoisomers. NMR experiments showed the presence of an anti-configuration of the methyl groups closest to the benzene ring and that the other methyl groups of the polyprenyl side-chain are not uniformly configured. Furthermore, socialane is structurally different from [6+2]-terpene viaticene of the closely related H. viatica, showing species specificity of the epicuticular lipids of this genus and hinting at a possible role of surface lipids in the communication of these gregarious arthropods.

The arylsulfatase- and phospholipase-rich venom of the plutoniumid centipede Theatops posticus.

Research on centipede venoms has led to the discovery of a diverse array of novel proteins and peptides, including those with homology to previously discovered toxin families (e.g., phospholipase A2s and pM12a metalloproteases) and novel toxin families not previously detected in venoms (e.g., ß-pore forming toxins and scoloptoxins). Most of this research has focused on centipedes in the order Scolopendromorpha, particularly those in the families Scolopendridae, Cryptopidae, and Scolopocryptopidae. To generate the first high-throughput venom characterization for a centipede in the scolopendromorph family Plutoniumidae, we performed venom-gland transcriptomics and venom proteomics on two Theatops posticus. We identified a total of 64 venom toxins, 60 of which were detected in both the venom-gland transcriptome and venom proteome and four of which were only detected transcriptomically. We detected a single highly abundant arylsulfatase B (ARSB) toxin, the first ARSB toxin identified from centipede venoms. As ARSBs have been detected in other venomous species (e.g., scorpions), ARSBs in T. posticus highlights a new case of convergent evolution across venoms. Theatops posticus venom also contained a much higher abundance and diversity of phospholipase A2 toxins compared to other characterized centipede venoms. Conversely, we detected other common centipedes toxins, such as CAPs and scoloptoxins, at relatively low abundances and diversities. Our observation of a diverse set of toxins from T. posticus venom, including those from novel toxin families, emphasizes the importance of studying unexplored centipede taxonomic groups and the continued potential of centipede venoms for novel toxin discovery and unraveling the molecular mechanisms underlying trait evolution.

Bioactive Peptides and Proteins from Centipede Venoms.

Venoms are a complex cocktail of biologically active molecules, including peptides, proteins, polyamide, and enzymes widely produced by venomous organisms. Through long-term evolution, venomous animals have evolved highly specific and diversified peptides and proteins targeting key physiological elements, including the nervous, blood, and muscular systems. Centipedes are typical venomous arthropods that rely on their toxins primarily for predation and defense. Although centipede bites are frequently reported, the composition and effect of centipede venoms are far from known. With the development of molecular biology and structural biology, the research on centipede venoms, especially peptides and proteins, has been deepened. Therefore, we summarize partial progress on the exploration of the bioactive peptides and proteins in centipede venoms and their potential value in pharmacological research and new drug development.

Centipede Venom: A Potential Source of Ion Channel Modulators.

Centipedes are one of the most ancient and successful living venomous animals. They have evolved spooky venoms to deter predators or hunt prey, and are widely distributed throughout the world besides Antarctica. Neurotoxins are the most important virulence factor affecting the function of the nervous system. Ion channels and receptors expressed in the nervous system, including NaV, KV, CaV, and TRP families, are the major targets of peptide neurotoxins. Insight into the mechanism of neurotoxins acting on ion channels contributes to our understanding of the function of both channels and centipede venoms. Meanwhile, the novel structure and selective activities give them the enormous potential to be modified and exploited as research tools and biological drugs. Here, we review the centipede venom peptides that act on ion channels.

Deoxybuzonamine Isomers from the Millipede Brachycybe lecontii (Platydesmida: Andrognathidae).

Millipedes (Diplopoda) are well known for their toxic or repellent defensive secretions. Here, we describe (6aR,10aS,10bR)-8,8-dimethyldodecahydropyrrolo[2,1-a]isoquinoline [trans-anti-trans-deoxybuzonamine (1a)] and (rel-6aR,10aR,10bR)-8,8-dimethyldodecahydropyrrolo[2,1-a]isoquinoline [trans-syn-cis-deoxybuzonamine (1b)], two isomers of deoxybuzonamine found in the chemical defense secretions of the millipede Brachycybe lecontii Wood (Colobognatha, Platydesmida, Andrognathidae). The carbon-nitrogen skeleton of these compounds was determined from their MS and GC-FTIR spectra obtained from the MeOH extract of whole millipedes, along with a subsequent selective synthesis. Their structures were established from their 1D (1H, 13C) and 2D NMR (COSY, NOESY, multiplicity-edited HSQC, HSQC-TOCSY, HMBC) spectra. Additionally, computational chemistry (DFT and DP4) was used to identify the relative configurations of 1a and 1b by comparing predicted 13C data to their experimental values, and the absolute configuration of 1a was determined by comparing its experimental specific rotation with that of the computationally calculated value. This is the first report of dodecahydropyrrolo[2,1-a]isoquinoline alkaloids from a platydesmidan millipede.

Potent Activity of Hybrid Arthropod Antimicrobial Peptides Linked by Glycine Spacers.

Arthropod antimicrobial peptides (AMPs) offer a promising source of new leads to address the declining number of novel antibiotics and the increasing prevalence of multidrug-resistant bacterial pathogens. AMPs with potent activity against Gram-negative bacteria and distinct modes of action have been identified in insects and scorpions, allowing the discovery of AMP combinations with additive and/or synergistic effects. Here, we tested the synergistic activity of two AMPs, from the dung beetle Copris tripartitus (CopA3) and the scorpion Heterometrus petersii (Hp1090), against two strains of Escherichia coli. We also tested the antibacterial activity of two hybrid peptides generated by joining CopA3 and Hp1090 with linkers comprising two (InSco2) or six (InSco6) glycine residues. We found that CopA3 and Hp1090 acted synergistically against both bacterial strains, and the hybrid peptide InSco2 showed more potent bactericidal activity than the parental AMPs or InSco6. Molecular dynamics simulations revealed that the short linker stabilizes an N-terminal 310-helix in the hybrid peptide InSco2. This secondary structure forms from a coil region that interacts with phosphatidylethanolamine in the membrane bilayer model. The highest concentration of the hybrid peptides used in this study was associated with stronger hemolytic activity than equivalent concentrations of the parental AMPs. As observed for CopA3, the increasing concentration of InSco2 was also cytotoxic to BHK-21 cells. We conclude that AMP hybrids linked by glycine spacers display potent antibacterial activity and that the cytotoxic activity can be modulated by adjusting the nature of the linker peptide, thus offering a strategy to produce hybrid peptides as safe replacements or adjuncts for conventional antibiotic therapy.

Arthropod Ectoparasites Have Potential to Bind SARS-CoV-2 via ACE.

Coronavirus-like organisms have been previously identified in Arthropod ectoparasites (such as ticks and unfed cat flea). Yet, the question regarding the possible role of these arthropods as SARS-CoV-2 passive/biological transmission vectors is still poorly explored. In this study, we performed in silico structural and binding energy calculations to assess the risks associated with possible ectoparasite transmission. We found sufficient similarity between ectoparasite ACE and human ACE2 protein sequences to build good quality 3D-models of the SARS-CoV-2 Spike:ACE complex to assess the impacts of ectoparasite mutations on complex stability. For several species (e.g., water flea, deer tick, body louse), our analyses showed no significant destabilisation of the SARS-CoV-2 Spike:ACE complex, suggesting these species would bind the viral Spike protein. Our structural analyses also provide structural rationale for interactions between the viral Spike and the ectoparasite ACE proteins. Although we do not have experimental evidence of infection in these ectoparasites, the predicted stability of the complex suggests this is possible, raising concerns of a possible role in passive transmission of the virus to their human hosts.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry: an emerging tool for studying the vectors of human infectious diseases.

Arthropod vectors have historically been identified morphologically, and more recently using molecular biology methods. However, both of these methods are time-consuming and require specific expertise and equipment. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry, which has revolutionized the routine identification of microorganisms in clinical microbiology laboratories, was recently successfully applied to the identification of arthropod vectors. Since then, the robustness of this identification technique has been confirmed, extended to a large panel of arthropod vectors, and assessed for detecting blood feeding behavior and identifying the infection status in regard to certain pathogenic agents. In this study, we summarize the state-of-the-art of matrix-assisted laser desorption ionization time-of-flight mass spectrometry applied to the identification of arthropod vectors (ticks, mosquitoes, phlebotomine sand-flies, fleas, triatomines, lice and Culicoides), their trophic preferences and their ability to discriminate between infection statuses.

R-hydroxynitrile lyase from the cyanogenic millipede, Chamberlinius hualienensis-A new entry to the carrier protein family Lipocalines.

Hydroxynitrile lyases (HNLs) catalyze the cleavage of cyanohydrin into cyanide and the corresponding aldehyde or ketone. Moreover, they catalyze the synthesis of cyanohydrin in the reverse reaction, utilized in industry for preparation of enantiomeric pure pharmaceutical ingredients and fine chemicals. We discovered a new HNL from the cyanogenic millipede, Chamberlinius hualienensis. The enzyme displays several features including a new primary structure, high stability, and the highest specific activity in (R)-mandelonitrile ((R)-MAN) synthesis (7420 U·mg-1 ) among the reported HNLs. In this study, we elucidated the crystal structure and reaction mechanism of natural ChuaHNL in ligand-free form and its complexes with acetate, cyanide ion, and inhibitors (thiocyanate or iodoacetate) at 1.6, 1.5, 2.1, 1.55, and 1.55 Å resolutions, respectively. The structure of ChuaHNL revealed that it belongs to the lipocalin superfamily, despite low amino acid sequence identity. The docking model of (R)-MAN with ChuaHNL suggested that the hydroxyl group forms hydrogen bonds with R38 and K117, and the nitrile group forms hydrogen bonds with R38 and Y103. The mutational analysis showed the importance of these residues in the enzymatic reaction. From these results, we propose that K117 acts as a base to abstract a proton from the hydroxyl group of cyanohydrins and R38 acts as an acid to donate a proton to the cyanide ion during the cleavage reaction of cyanohydrins. The reverse mechanism would occur during the cyanohydrin synthesis. (Photo: Dr. Yuko Ishida) DATABASES: Structural data are available in PDB database under the accession numbers 6JHC, 6KFA, 6KFB, 6KFC, and 6KFD.

3D-Printed Bioinspired Cassie-Baxter Wettability for Controllable Microdroplet Manipulation.

It is a great challenge to fabricate a surface with Cassie-Baxter wettability that can be continuously adjusted from hydrophilicity to superhydrophobicity by changing of geometric parameters. In this paper, we propose and demonstrate a bioinspired surface fabricated by using a projection micro-stereolithography (PµSL) based 3D printing technique to address the challenge. Independent of materials, the bioinspired textured surface has a maximum contact angle (CA) of 171°, which is even higher than that of the omniphobic springtail skin we try to imitate. Most significantly, we are able to control the CA of the bioinspired surface in the range of 55-171° and the adhesion force from 71 to 99 µN continuously by only changing the geometric parameters of the bioinspired microstructures. The underlying mechanisms of the CA control of our bioinspired surface are also revealed by using a multi-phase lattice Boltzmann model. Furthermore, we demonstrate potential applications in droplet-based microreactors, nonloss water transportation, and coalescence of water droplets by employing our 3D-printed bioinspired structures with their remarkable precise Cassie-Baxter wettability control and petal effects. The present results potentially pave a new way for designing next generation functional surfaces for microdroplet manipulation, droplet-based biodetection, antifouling surfaces, and cell culture.

A Review of Pedal Peptide/Orcokinin-type Neuropeptides.

Neuropeptides are endogenous active substances that play important roles in a number of physiological processes and are ubiquitous in the nervous tissue in vivo. The gene encoding pedal peptide/orcokinin-type (PP/OK-type) neuropeptide is an important member of the neuropeptide gene family and is ubiquitous in invertebrates of Bilateria; orcokinin (OK) is mainly found in Arthropoda, while pedal peptide (PP) is mainly found in Mollusca. OK and PP are also present in other animals. PP/OK-type neuropeptides are a kind of multifunctional neuropeptides predominantly expressed in the nervous tissue and play important roles in the nerve regulation of movement. Moreover, OK has a number of other physiological functions. This review describes the distribution, expression, function and maturation of PP/OK-type neuropeptides to facilitate investigations of new functions and receptors of PP/OK-type neuropeptides, providing the theoretical foundation for the potential use of PP/OK-type neuropeptides in the prevention and control of agricultural and forestry pests, as an additive for skin care products and in the screening of drugs for the treatment of diabetes.

Wild Blonde Capuchins (Sapajus flavius) Perform Anointing Behaviour Using Toxic Secretions of a Millipede (Spirobolida: Rhinocricidae).

Defensive secretions of millipedes are remarkable for containing toxic quinones known to efficiently repell hematophagous arthropods. Here we show that Endangered blonde capuchin monkeys make use of such secretions. We (i) describe the anointing behavior performed by the monkeys (ii) identify the millipede species used in the process (iii) describe the volatile chemical composition of its secretion. The blonde capuchin monkeys selectively searched for millipedes hidden under the ground. We observed three bouts of anointing behavior, performed by 13 individuals of all age classes (from adults to independent infants), both solitarily (1 event) and socially (10 events). The only millipede species used by the monkeys is an undescribed species of the genus Poecilocricus (Spirobolida, Rhinocricidae). The volatile chemical composition of the secretions was predominantly comprised of a mixture of benzoquinones and hydroquinones. The social nature of the behavior and time of the observations (mosquito season), suggest that social bonding and mosquito avoidance is linked to the anointing behavior of the monkeys.

Gosodesmine, a 7-Substituted Hexahydroindolizine from the Millipede Gosodesmus claremontus.

Millipedes (Diplopoda) are well known for their toxic or repellent defensive secretions. Here we describe gosodesmine (1), 7-(4-methylpent-3-en-1-yl)-1,2,3,5,8,8a-hexahydroindolizine, a unique alkaloid with some terpene character found in the chemical defense secretions of the millipede Gosodesmus claremontus Chamberlin (Colobognatha, Platydesmida, Andrognathidae). The structure of 1 was suggested by its mass spectra and GC-FTIR spectra and established from its 1H, 13C, and 2D NMR spectra and 1D NOE studies. The 7-substituted indolizidine carbon skeleton of 1 was confirmed by unambiguous synthesis. This is the first report of an alkaloid from a platydesmid millipede and the first report of a 7-substituted indolizidine from an arthropod.

Development of alginate beads with encapsulated jabuticaba peel and propolis extracts to achieve a new natural colorant antioxidant additive.

This work aims to encapsulate anthocyanins and phenolic compounds extracted from a native Brazilian fruit peel - jabuticaba (Plinia cauliflora (Mart.) Kausel) and propolis from Tubuna (Scaptotrigona bipunctata) stingless bees, with great potential benefits for human health. The alginate encapsulation was conducted by the ionotropic gelation through the dripping into the CaCl2 solution. Both raw extracts were characterized by TPC - total phenolic content (Folin-Ciocalteu), AA -antioxidant activity (DPPH and ABTS assays), and TMAC - total monomeric anthocyanin concentration (pH differential method); as well as their resultant mixture (2:1 jabuticaba/propolis). The obtained beads presented highly efficient encapsulation of total polyphenols (~98%) and monomeric anthocyanins (~89%), with spherical morphology and smooth surface obtaining a mean diameter between 200 and 250 µm. In vitro release study showed that JPE/alginate beads were completely disintegrated at pH 7.4 (intestinal pH), but they were resistant to gastric pH (1.2) presenting a slow release of about 40% in 240 min. This is the first report that encapsulates the mixture of jabuticaba and propolis extracts and may contribute to the utilization of a great source of bioactive compounds besides the potential pigment of anthocyanins, an alternative to natural and healthy food/beverage colorants.

Shifting systems: prerequisites for the application of quantitative fatty acid signature analysis in soil food webs.

Quantitative fatty acid signature analysis (QFASA) is widely used to investigate trophic interactions in marine ecosystems, as nutritionally important ω3 long-chain polyunsaturated fatty acids at the food web base allow tracing of their trophic transfer in the food chain. By contrast, the basal resources in soil food webs comprise a wider array of trophic markers, including branched-chain, cyclopropane as well as several mono- and polyunsaturated fatty acids. These diverse markers allow distinguishing between the three dominant soil carbon and energy channels, the root, bacterial and fungal pathway. QFASA has not been applied yet to soil ecosystems owing to the lack of a priori data to fit the model. The present work investigates the transfer of absolute and relative trophic marker fatty acids into Collembola as dominant representatives of the soil mesofauna. Three different species were fed on a variety of single diets characteristic for the green and brown food chain. Calibration coefficients were calculated and diet estimation trials for mixed diet set-ups were performed, using a library comprising 50 different resources. However, estimation of Collembola diet was only partially successful, identifying the main components, but not the correct relative proportions. Adjustments by fat content or diet group exclusion did not improve the results. Nonetheless, this work provides, to our knowledge, a first comprehensive dataset to translate the application of QFASA from marine to soil ecosystems. This article is part of the theme issue 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.

Reliable and Robust Fabrication Rules for Springtail-Inspired Superomniphobic Surfaces.

We report a reliable and robust method for the fabrication of bioinspired superomniphobic surfaces with precise concave-cap-shaped micropillar arrays. This method includes silicon-based conventional microelectromechanical systems (MEMS) and polymer replication processes. We have elucidated two critical cases of fabrication rules for precise micromachining of a negative-shaped bioinspired silicon master. The fabricated polymeric structure replicated from the semipermanent silicon master based on the design rules exhibited high structural fidelity and robustness. Finally, we validated the superomniphobic properties, structural durability, and long-term stability of the fabricated bioinspired surfaces.

Antibacterial action of lactoferricin B like peptide against Escherichia coli: reactive oxygen species-induced apoptosis-like death.

AIMS: Emergence and rapid dissemination of antibiotic-resistant bacteria is becoming a severe problem to public health. The search for antimicrobial substitutes for antibiotics is necessary. Lactoferricin B like peptide (LBLP) is a 23-mer antimicrobial peptide (AMP), derived from the big centipede Scolopendra subspinipes mutilans. Although its antifungal effect and its mechanism have been reported, the antibacterial activity has not yet been elucidated. METHOD AND RESULTS: In this study, we investigated antibacterial activity of LBLP and its mode of action. LBLP showed potent antibacterial effect against pathogenic bacteria Escherichia coli and did not show haemolytic activity against human erythrocyte. The general antimicrobial mechanism of AMP is to disrupt the cell membrane, however, LBLP exerted its antibacterial activity by causing apoptosis-like death through reactive oxygen species (ROS) generation. LBLP-treated E. coli cells exhibited hallmarks of apoptosis, such as membrane depolarization, DNA fragmentation, caspase-like protein activation and phosphatidylserine exposure. These apoptotic features were attenuated by pretreatment of NAC, a representative ROS scavenger. CONCLUSIONS: These results demonstrate that LBLP exerted its antibacterial activity by generating ROS and inducing apoptosis-like death in E. coli. LBLP is not membrane destructive per se, but essentially a metabolic inhibitor. SIGNIFICANCE AND IMPACT OF THE STUDY: Lactoferricin B like peptide is potential candidate to replace conventional antibiotics that are less effective because of its unique properties.

Inhibition of Candida albicans and Staphylococcus aureus biofilms by centipede oil and linoleic acid.

Microbial biofilms are associated with persistent infections because of their high tolerance to antimicrobial agents and host defenses. The effects of centipede oil from Scolopendra subspinipes mutilans and its main components were investigated to identify non-toxic biofilm inhibitors. Centipede oil and linoleic acid at 20 µg ml-1 markedly inhibited biofilm formation by two fluconazole-resistant Candida albicans strains and three Staphylococcus aureus strains without affecting their planktonic cell growth. Also, both centipede oil and linoleic acid inhibited hyphal growth and cell aggregation by C. albicans. In addition, centipede oil and linoleic acid showed anti-biofilm activities against mixed C. albicans and S. aureus biofilms. Transcriptomic analysis showed that centipede oil and linoleic acid downregulated the expressions of several hypha/biofilm-related genes in C. albicans and α-hemolysin in S. aureus. Furthermore, both compounds effectively reduced C. albicans virulence in a nematode infection model with minimal toxicity.

Reisolation and Configurational Reinvestigation of Cottoquinazolines E-G from an Arthropod-Derived Strain of the Fungus Neosartorya fischeri.

The reported fumiquinazoline-related alkaloids cottoquinazolines E-G (1-3) were reisolated from solid cultures of the fungus Neosartorya fischeri, which was isolated from the medicinal arthropod Cryptotympana atrata. The unresolved issues regarding the absolute configurations (for cottoquinazolines E and F) prompted a reinvestigation of the configurations for all three compounds, as enabled by extensive spectroscopic methods, comparisons of experimental electronic circular dichroism data, and X-ray crystallography. In addition, cottoquinazoline F (2) showed significant antibacterial activity against ESBL-producing Escherichia coli, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterococcus faecalis with MIC values of 8, 32, 32, and 16 µg/mL, respectively.