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Dilaridae are an ancient relict lineage of lacewings, whose life history is poorly known. We investigated the external morphology of the first and second instar larvae of Dilar duelli Aspock U. and Aspock H., 1995, based on reared and collected specimens by means of scanning electron microscopy. Larvae of Dilar are characterized by straight mandibulo-maxillary stylets, strengthened antennae and palps, absence of stemmata, robust prothoracic legs with asymmetrical pretarsal claws, greatly elongate and cylindrical abdomen, and large pygopods, all of these features appear to be adaptations to fossorial habits, allowing the larvae to move in the soil. Dilar undergoes drastic developmental changes between larval instars, changing from minute, stout first instars to long, slender third instars, characterized by an overly developed abdomen. Larvae of Dilaridae share several morphological and developmental features with Mantispidae and Berothidae, although phylogenetic evidence suggests that these traits arose independently in association with similar life habits.

Life cycle of the aquatic moth Cataclysta lemnata (Lepidoptera: Crambidae) was studied in laboratory conditions to obtain a basic biological knowledge useful for predicting the possible success of the herbivorous larvae of this insect as potential control agents in limiting the spread of the invasive American duckweed Lemna minuta (Alismatales: Araceae) in Italy. The multivoltinism of C. lemnata, as well as the high overall emergence from the pupal stage (85%), the high success in mating among the formed couples (>90%), and the high number of larvae born from each egg laying (on average 310 individuals), suggest that the insect can be successfully bred in the laboratory for the purposes of an augmentative biological control. Under experimental conditions, larvae developed in 23 days (through six larval instars, distinguishable by cephalic capsule dimensions) and pupae in 10, with no difference in duration between females and males. The larval phase resulted longer than the adult one (23 vs 10 days), therefore it can be considered the most suitable stage for releasing the insect in field for biocontrol purposes. Indeed, the larvae having a herbivorous diet might consume large amount of the invasive plant, contrarily to the adult phase which is focused exclusively on reproduction. The results emerged not only allow to contribute to the knowledge on aquatic lepidoptera that are scarcely known, but also support the effectiveness of a possible protocol for an augmentative biological control of the invasive alien duckweed L. minuta.

The morphology of larval instars I, II and III of the water beetle Ochthebius (Cobalius) balfourbrownei Jach, 1989 from Cape Verde Islands is described and illustrated for the first time, by scanning electron microscopy, with a special emphasis on their chaetotaxy and porotaxy. Specimens used in this study were collected with adults of the same species in marine rockpools. A detailed description of characters is provided for comparison with the other members of the same genus inhabiting the marine rockpools, and illustrations of the diagnostic features are presented, with the aim of providing information useful for taxonomic and phylogenetic studies of the genus Ochthebius.

Several works have investigated the impact of urbanisation on carabid activity density using urban-rural gradients. Such works compared activity density recorded from green spaces located in different parts of a city and assigned to categories of increasing urban intensity, which poses two problems: (1) since the gradient is divided into categories, it is impossible to model continuous variations in biotic responses, and (2) sites representative of different urbanisation levels are not true segments of the same ecological continuum. To surpass these problems, we modelled variations in carabid activity density along an urban-rural transect within a single green space extending from the city centre of Rome to rural environments. Carabids were sampled by pitfall traps from sites distributed along the entire gradient. We used breakpoint regressions to model how (1) carabid activity density, (2) carabids/beetles ratio, (3) carabids/insects ratio and (3) carabids/arthropods ratio varied along the gradient. As already observed for various organisms in urban environments, we found that activity density of carabids and their contribution to the abundance of beetles, insects and arthropods, peaked in the middle of the gradient. This supports the intermediate disturbance hypothesis, according to which moderate urbanisation may favour diversity by increasing habitat heterogeneity.

The peculiar morphology of the discotelic larvae of three species of Pachyteles (Coleoptera, Carabidae, Paussinae, ozaenini) collected in an Andean cloud forest in Ecuador (otonga nature Reserve, Cotopaxi) is here analyzed. All instars of Pachyteles vignai Deuve, 1999, 2nd and 3rd instars of P. digiulioi Deuve, 1999, and the 3rd instar of another undetermined Pachyteles species are described. A particular emphasis on the illustration of the aberrant larval morphology is given by using both traditional drawing techniques and macro multifocal image acquisition, combined with scanning electron microscopy (SEM) for analyzing the details. The diagnostic characters of these larvae are compared to those of the other known larvae of ozaenini.

On the basis of the materials collected on site, the larvae of six species of Mexican Calosoma, namely Calosoma (Callitropa) porosifrons Bates, 1891, and five species of the subgenus Carabomimus Kolbe, 1895 are figured and described for the first time. Though these larvae are quite similar for most characters, some important differences can be found in the two subgenera Callitropa Motschulsky, 1865 and Carabomimus, confirming the distinctness of these taxa. Through an accurate study of the collected adult specimens and of the relevant literature, the distribution of the population afferent to each of these same species is clarified. With the occasion, the distribution of all Mexican Calosoma, is discussed, distinguishing between winged species, usually widely spread, and apterous species, linked to humid, temperate or semi-cold climates. In this context, the distribution of the subgenus Carabomimus, appears to be more sensitive to the influence of geographic barriers and the differentiation of the species was clearly helped by geographic isolation at high altitude in the volcanic massif of central Mexico.

Background Autoimmune and multifocal atrophic gastritis (AG) are at risk for gastric neoplastic lesions. European guidelines recommend surveillance with high-quality endoscopy every 3 years. Aim To prospectively investigate the occurrence of gastric neoplastic lesions at the 3-year follow-up in patients with autoimmune and multifocal AG. Methods Longitudinal cohort study conducted between 2011 and 2019: consecutive patients with histological diagnosis of autoimmune or multifocal AG underwent follow-up gastroscopy 3 years after diagnosis with high-resolution-narrow-band-imaging endoscopes. Results Overall, 160 patients were included(F117(73.0%);median age 66(35–87)years). Autoimmune and multifocal AG were present in 122(76.3%) and 38(23.7%) patients, respectively. At the 3-year follow-up, 16(10.0%) patients presented 16 gastric neoplastic lesions: 3(18.7%) gastric cancers, 4(25.0%) low-grade dysplasia, 2(12.5%) low-grade dysplasia adenomas, 7(43.7%) type-1 neuroendocrine tumours. In these patients, OLGA and OLGIM III/IV stages were present in 4(25.0)% and 1(6.3%), respectively; 11(69.0%) presented autoimmune AG, and all but one(93.7%) had parietal cells antibodies positivity (p = 0.026 vs patients without lesions). All lesions were endoscopically(87.5%) or surgically(12.5%) treated with favourable outcome. Age>70 years was associated with a 9-fold higher probability of developing gastric epithelial neoplastic lesions (OR 9.6,95CI% 1.2–79.4,p = 0.0359). Conclusions The first endoscopic surveillance 3 years after diagnosis seems safe for autoimmune and multifocal AG patients and should be offered to elderly patients who are at higher risk for gastric neoplasia.

The southern branch of the Elizabeth River near Portsmouth, Virginia, USA, is one of the most creosote-polluted subestuaries in North America and the former location of the Atlantic Wood US Environmental Protection Agency Superfund site. We previously demonstrated that adult Atlantic Wood killifish collected in situ had severe hepatic lesions, including hepatoblastoma and hepatocellular carcinoma, as well as suppressed circulating antibody responses compared to a historical reference site. Moreover, several innate immune functions were higher in Atlantic Wood fish, including elevated expression of hepatic cyclooxygenase-2 (COX-2), suggesting a proinflammatory environment. To further examine the potential of Atlantic Wood contaminants to modulate innate immune function(s), the present study used RAW264.7 mouse macrophages as an in vitro model to develop new approach methodologies for rapid screening. Lipopolysaccharide (LPS)-stimulated nitric oxide secretion by macrophages is a rapid, sensitive, and predictive in vitro system for screening potentially immunotoxic contaminants as single compounds or as complex mixtures. Compared to the reference site, filter-sterilized Atlantic Wood sediment extracts (water accommodated fractions) induced nitric oxide and IL-6 secretion as well as inducible nitric oxide synthase and COX-2 proteins at levels comparable to or higher than those induced by LPS treatments alone. Extracts also increased phagocytic activity by macrophages. Using a limulus lysate assay, we show that bacterial endotoxin levels in Atlantic Wood extracts are higher than in reference extracts and that polymyxin-B chelation ameliorates proinflammatory effects. These findings illuminate the reality of sediment constituents other than toxic compounds previously associated with developmental abnormalities and carcinogenesis in killifish from the Atlantic Wood site. Perhaps these data also suggest the presence of contaminant-adapted consortia of sediment microbes at many heavily polluted sites worldwide compared to less contaminated sites. Environ Toxicol Chem 2021;40:1576-1585. © 2021 SETAC.

Background Meloidae (blister beetles) are known to synthetize cantharidin (CA), a toxic and defensive terpene mainly stored in male accessory glands (MAG) and emitted outward through reflex-bleeding. Recent progresses in understanding CA biosynthesis and production organ(s) in Meloidae have been made, but the way in which self-protection is achieved from the hazardous accumulation and release of CA in blister beetles has been experimentally neglected. To provide hints on this pending question, a comparative de novo assembly transcriptomic approach was performed by targeting two tissues where CA is largely accumulated and regularly circulates in Meloidae: the male reproductive tract (MRT) and the haemolymph. Differential gene expression profiles in these tissues were examined in two blister beetle species, Lydus trimaculatus (Fabricius, 1775) (tribe Lyttini) and Mylabris variabilis (Pallas, 1781) (tribe Mylabrini). Upregulated transcripts were compared between the two species to identify conserved genes possibly involved in CA detoxification and transport. Results Based on our results, we hypothesize that, to avoid auto-intoxication, ABC, MFS or other solute transporters might sequester purported glycosylated CA precursors into MAG, and lipocalins could bind CA and mitigate its reactivity when released into the haemolymph during the autohaemorrhaging response. We also found an over-representation in haemolymph of protein-domains related to coagulation and integument repairing mechanisms that likely reflects the need to limit fluid loss during reflex-bleeding. Conclusions The de novo assembled transcriptomes of L. trimaculatus and M. variabilis here provided represent valuable genetic resources to further explore the mechanisms employed to cope with toxicity of CA in blister beetle tissues. These, if revealed, might help conceiving safe and effective drug-delivery approaches to enhance the use of CA in medicine.

Chloroquine (CQ) and hydroxychloroquine (HCQ) are derivatives of the heterocyclic aromatic compound quinoline. These economical compounds have been used as antimalarial agents for many years. Currently, they are used as monotherapy or in conjunction with other therapies for the treatment of autoimmune diseases such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), Sjögren's syndrome (SS) and antiphospholipid antibody syndrome (APS). Based on its effects on the modulation of the autophagy process, various clinical studies suggest that CQ and HCQ could be used in combination with other chemotherapeutics for the treatment of various types of cancer. Furthermore, the antiviral effects showed against Zika, Chikungunya, and HIV are due to the annulation of endosomal/lysosomal acidification. Recently, CQ and HCQ were approved for the U.S. Food and Drug Administration (FDA) for the treatment of infected patients with the coronavirus SARSCoV- 2, causing the disease originated in December 2019, namely COVID-2019. Several mechanisms have been proposed to explain the pharmacological effects of these drugs: 1) disruption of lysosomal and endosomal pH, 2) inhibition of protein secretion/expression, 3) inhibition of antigen presentation, 4) decrease of proinflammatory cytokines, 5) inhibition of autophagy, 6) induction of apoptosis and 7) inhibition of ion channels activation. Thus, evidence has shown that these structures are leading molecules that can be modified or combined with other therapeutic agents. In this review, we will discuss the most recent findings in the mechanisms of action of CQ and HCQ in the immune system, and the use of these antimalarial drugs on diseases.

Chronic wound management becomes a complex procedure because of the persistence of forming biofilm pathogens that do not respond to antimicrobial treatment. The aim of this paper is to detect the Graphene Oxide-GO effect on Staphylococcus aureus and Pseudomonas aeruginosa dual species wound biofilm in Lubbock Chronic Wound Biofilm-LCWB model. LCWB is a recognized model that mimics the spatial microbial colonization into chronic wounds and reproduces the wound and its clot. Staphylococcus aureus PECHA 10 and P. aeruginosa PECHA 4, are the pathogens used in the study. The GO effect on both in forming and mature biofilms, is detected by the evaluation of the CFU/mg reduction, the cell viability and ultrastructural analysis of the treated LCWBs. Graphene Oxide, at 50 mg/l, shows a significant antibiofilm effect in forming and mature LCWBs. In particular, during the biofilm formation, GO reduces the S. aureus and P. aeruginosa growth of 55.05% ± 4.73 and 44.18% ± 3.91 compared to the control. In mature biofilm, GO affects S. aureus and P. aeruginosa by reducing their growth of 70.24% ± 4.47 and 63.68% ± 17.56, respectively. Images taken by SEM show that GO display a disaggregated microbial effect also disrupting the fibrin network of the wound-like biofilm framework. In conclusion, GO used against microorganisms grown in LCWB, displays a significant inhibitory action resulting in a promising tool for potential application in wound management.

Emerging evidence suggests the role of environmental chemicals, in particular endocrine-disrupting chemicals (EDCs), in progression of breast cancer and treatment resistance, which can impact survival outcomes. However, most research tends to focus on tumor etiology and the effect of single chemicals, offering little insight into the effects of realistic complex mixture exposures on tumor progression. Herein, we investigated the effect of a polycyclic aromatic hydrocarbon (PAH)-enriched EDC mixture in a panel of normal and breast cancer cells and in a tumor organoid model. Cells or organoids in culture were treated with EDC mixture at doses estimated from US adult intake of the top four PAH compounds within the mixture from the National Health and Nutrition Examination Survey database. We demonstrate that low-dose PAH mixture (6, 30 and 300 nM) increased aryl hydrocarbon receptor (AhR) expression and CYP activity in estrogen receptor (ER) positive but not normal mammary or ER-negative breast cancer cells, and that upregulated AhR signaling corresponded with increased cell proliferation and expression of antiapoptotic and antioxidant proteins XIAP and SOD1. We employed a mathematical model to validate PAH-mediated increases in AhR and XIAP expression in the MCF-7 ER-positive cell line. Furthermore, the PAH mixture caused significant growth increases in ER-negative breast cancer cell derived 3D tumor organoids, providing further evidence for the role of a natural-derived PAH mixture in enhancing a tumor proliferative phenotype. Together, our integrated cell signaling, computational and phenotype analysis reveals the underlying mechanisms of EDC mixtures in breast cancer progression and survival.

I analyzed the model that suggests that an RNA ring might have been the progenitor of the tRNA molecule (Demongeot and Moreira in J Theor Biol 249:314-324, 2007; Demongeot and Seligmann in J Mol Evol 1-23, 2019a; Demongeot and Norris in Life 9(2):51, 2019). In particular, I analyze three ways in which this precursor, especially in its RNA hairpin form, could have evolved into the complete tRNA molecule. These three modalities are based on multiple duplication events, and therefore, appear to be less parsimonious than that which assumes that this molecule originated through one duplication of a single hairpin structure. The conclusion is, therefore, that the latter model appears to be preferable with respect to that of the RNA ring, also because there are many independent observations and some of a historical nature that would corroborate it in an extraordinary way.

The wood species of 18th- and 19th-century Italian historic berlines (gala carriages mainly made of wood) housed in the Palazzo Pitti museum in Florence were investigated using non-destructive techniques based on the use of portable digital microscopes. Overall, thirteen wood taxa were identified. Among those, walnut (Juglans regia) was the most abundant species in berlines commissioned by noblemen of high rank, such as kings, princes and grand dukes, whereas in the other carriages of lower lineage, the walnut was replaced by elm (Ulmus sp.) wood, a species commonly used for carriages. Some species were typically present in specific parts of the structures, such as elm and ash (Fraxinus sp.) in the transoms and struts of the undercarriage, oak (Quercus sp.) in the wheel's spokes, poplar (Populus sp.) in the seats, panels and mouldings of the body and European beech (Fagus sylvatica) in the light frames. Species found only occasionally, including chestnut (Castanea sativa), fir (Abies alba) and spruce (Picea abies), may have been introduced in occasions of restoration or substitution. The results also showed a preference for the use of local species, which is different from carriages in other European countries where exotic species were already used.

As disease worsens in patients with cystic fibrosis (CF), Pseudomonas aeruginosa (PA) colonizes the lungs, causing pulmonary failure and mortality. Progressively, PA forms typical biofilms, and antibiotic treatments determine multidrug-resistant (MDR) PA strains. To advance new therapies against MDR PA, research has reappraised bacteriophages (phages), viruses naturally infecting bacteria. Because few in vitro studies have tested phages on CF PA biofilms, general reliability remains unclear. This study aimed to test in vitro newly isolated environmental phage activity against PA isolates from patients with CF at Bambino Gesù Children’s Hospital (OBG), Rome, Italy. After testing in vitro phage activities, we combined phages with amikacin, meropenem, and tobramycin against CF PA pre-formed biofilms. We also investigated new emerging morphotypes and bacterial regrowth. We obtained 22 newly isolated phages from various environments, including OBG. In about 94% of 32 CF PA isolates tested, these phages showed in vitro PA lysis. Despite poor efficacy against chronic CF PA, five selected-lytic-phages (Φ4_ZP1, Φ9_ZP2, Φ14_OBG, Φ17_OBG, and Φ19_OBG) showed wide host activity. The Φ4_ZP1-meropenem and Φ14_OBG-tobramycin combinations significantly reduced CF PA biofilms (p &lt, 0.001). To advance potential combined phage-antibiotic therapy, we envisage further in vitro test combinations with newly isolated phages, including those from hospital environments, against CF PA biofilms from early and chronic infections.

Pelamera atra (Rondani, 1861) is an enigmatic species of bristle fly that has been challenging dipterologists with regard to its taxonomic position within the family Tachinidae since it was first described. The species is rarely collected and is only known from a handful of female specimens. The first ever male specimen of P. atra was collected in southern Italy in 2017, and it allowed for the discovery of its unique terminalia. The present paper provides the first description of a male of P. atra and presents new insights about its phylogenetic position within the family based on molecular data. A maximum likelihood analysis was conducted on a selection of tachinid taxa using sequences of two nuclear protein-coding genes (CAD and MCS) to formulate the first hypothesis concerning the phylogenetic placement of P. atra. Phylogenetic analyses reconstructed P. atra as a member of the Tachininae sister to the (Loewiini + (Polideini + Ernestiini)) clade. This reconstruction challenges previous attempts at classifying Pelamera based on female external morphology, which instead suggested a close affinity to the tribes Brachymerini or Minthoini. Due to the morphological distinctiveness of this taxon, we here propose the placement of P. atra in the monotypic tribe Pelamerini stat. rev.

Hastisetae are modified setae typical of Dermestidae Megatominae and are a primary defensive tool of both larvae and pupae against invertebrates and possibly vertebrates. Given their unique morphological features, hastisetae have recently been suggested as an additional character useful for larvae identification and possible source of information to clarify the systematics of Megatominae. Hastisetae are also recognized as important contaminants of stored products, work and living environment, in particular, the exposure to hastisetae seems to cause allergic reactions and the insurgence of skin rashes, asthma, conjunctivitis, and digestive system inflammation in humans. Starting from these basic concepts, the present paper provides a detailed description of the hastisetae of some Megatominae. Fine morphology of external and internal microstructures of the hastisetae is shown and compared at the genus level. The insertion on integument, the pedicel, the shaft, and the apical head are illustrated in detail, and the first observations of active defensive behavior based on hastisetae are recorded and presented. Possible implications to the systematics of skin beetles are proposed based on the results of the study.

Larvae of the two European species of the genus Pyrochroa (P. coccinea and P. serraticornis) are redescribed, for the first time based on Italian samples, and new relevant and distinctive characters have emerged from P. serraticornis. Particularly, the presence of a single urogomphal pit between urogomphi (representing a new condition for the entire family), and new characters of mandibles, open new questions about the current taxonomic status of the European species.

We explored the structure of plant assemblages that settles around the anthills of a guild of Hymenoptera Formicidae (Messor wasmanni Krausse 1910, Tapinoma nigerrimum Nylander 1856 and Aphaenogaster spinosa Emery 1878), observable in urban grasslands dominated by Dasypyrum villosum (Rome, Central Italy). Since it is known that ants act as a disturbing factor on plant assemblages of grasslands, our hypothesis was that vegetation structures suffer from some stressors that affect, in this plant association, the dominant plant structure. We compared the plant assemblages observed in the plots of the anthills with the control grassland assemblages using a diversity/dominance diagram. We recorded 63 plant taxa. The average number of plant species was found to be significantly lower in anthill plots than in control grassland plots. In anthill plots, dominant plant species (Polygonum romanum, Poa trivialis Vulpia myuros, Aira elegantissima and Vulpia ligustica) have been found to be different from control grassland plots (Convolvolus arvensis, Dasypyrum villosum, Poa trivialis and Sheradia arvensis). Anthill plant assemblages were found to be significantly different (One-way PERMANOVA) and poorer in terms of species richness, less diversified, and with a reduced species turnover than control grassland plots. Whittaker plot analysis seems to show that the plant assemblages of the control, with greater evenness, emphasize a stressed condition in anthill plant assemblages. Non-metric multidimensional scaling shows a set of species strictly linked to anthills. Our data seem to confirm the role of ants as a stressor in the plant assemblage structures of Mediterranean urban grasslands. However, our results also show that the peculiar ant nest conditions may favor plant species less represented in the Dasypyretum grasslands (i.e. Polygonum romanum) or even absent in this plant assemblage (i.e. Aira elegantissima, Cynodon dactylon and Poa annua), thus increasing the overall local plant diversity.

Fossils sometimes show unusual morphological features absent in living organisms, making it difficult to reconstruct both their affinity and their function. We describe here a newlacewing larva, Ankyloleon caudatus gen. et sp.n. (Neuroptera) from the Cretaceous amber of Myanmar, characterized by an abdomen unique among insects, with ‘tail-like’ terminal segments bearing a ventral pair of vesicles. Phase-contrast X-ray microtomography reveals that these structures were dense and equipped with a median duct, suggesting that they were likely pygopods used for locomotion, holding the position through adhesive secretions. Our phylogenetic analyses, combining genomic and morphological data from both living and fossil lacewings, proved critical to placing Ankyloleon gen.n. on the lacewing tree of life as an early representative of the antlion clade, Myrmeleontiformia. These results corroborate the view that derived myrmeleontiform lacewings ‘experimented’ with unusual combinations of features and specializations during their evolutionary history, some of which are now lost.

Simple Summary The present paper is aimed to further explore the structure of the antennae and mouthparts of specialized beetle species living on flowers, as well as the functions of their associated sensilla. In this study, we used scanning electron microscopy (SEM) to observe and describe for the first time the fine morphology of sensilla on the antennae and mouthparts of the pollen-beetle Meligethes (Odonthogethes) chinensis, a common Chinese species associated with flowers of Rosaceae. The results show that there are six types and twelve subtypes of sensilla on male antennae; seven types and fourteen subtypes on female antennae; seven types and seventeen subtypes on male mouthparts; seven types and sixteen subtypes on female mouthparts. The sensilla on the antennae and mouthparts of Meligethinae that feed on pollen were finally compared with similar sensilla known to occur in other insects, in order to obtain more insights on the evolution of these sensorial structures in specialized flower-inhabiting insects. Abstract Meligethes (Odonthogethes) chinensis is a common Chinese phytophagous species in the family Nitidulidae. Its main larval host plant is Rubus idaeus L. (Rosaceae), and adults feed on pollen and other floral parts. In this study, we used scanning electron microscopy (SEM) to observe and study the fine morphology of sensilla on the antennae and mouthparts of M. chinensis. The results show that there are six types and twelve subtypes of sensilla on male antennae; seven types and fourteen subtypes on female antennae; seven types and seventeen subtypes on male mouthparts; seven types and sixteen subtypes on female mouthparts. Sensilla coeloconica (SCo) are found on the female antennae of M. chinensis only, and they are also reported on the antennae of Nitidulidae for the first time. SCo2 on the labrum present sexual dimorphism, and one subtype of sensilla basiconica (SB6) is presented on the tip of maxillary and labial palps of the male only, while other types of sensilla are very similar on the mouthparts of male and female. Finally, by comparing similar sensilla in other insects, we also attempted to discuss the functions of all sensilla on the antennae and mouthparts of M. chinensis.

The hoverfly Microdon myrmicae is a rare and extremely localised social parasite of Myrmica ants, only occurring around wet grassland. Dispersal, location of the host, and oviposition are crucial steps in the life of ant parasites but are poorly known due to the challenge of studying such rare species. Using genome-wide loci obtained by RADseq, we investigated the genetic structure and relatedness of Mi. myrmicae larvae and its ant host, collected from three localities in South West England, and inferred biological and behavioural traits. We found that: (a) Mi. myrmicae larvae show high inbreeding levels and severe heterozygosity deficiency, as an expression of the small population size that favours the mating between siblings or half-siblings; (b) Mi. myrmicae adults can disperse for many kilometres, spreading much more than it is reported for its sibling species, Microdon mutabilis; (c) a single female lays small egg bunches in different ant colonies, sometimes spanning substantial distances (bet-hedging strategy); in parallel, a single ant nest can harbour eggs from different Microdon females; (d) preliminary evidence suggest that contrary to Mi. mutabilis, host colony choice seems not to depend on the number of queens residing in a single colony. These results overall strongly deviate from what previously found for Mi. mutabilis, whose females oviposit in the natal nest generation after generation. We argue that such different ecological traits and parasitic strategies between closely related species are mainly ascribable to the different selective pressure on the two ant hosts.

The stage reached by the evolution of cellularity in the Last Universal Common Ancestor (LUCA) has not yet been identified. In actual fact, it has not been clarified whether the LUCA was a cell (genote) or a protocell (progenote). Recently, Pende et al. (2021) analysed the phylogenetic distribution of the cell division system present in bacteria and archaea reaching the conclusion that LUCA was a cell and not a progenote. I find this conclusion unreasonable with respect to the observations they presented. One of the points is that the presence in the domains of life of many genes - some paralogs - which would define the membrane-remodeling superfamily would seem to imply a tempo and a mode of evolution for the LUCA more typical of the progenote than the genote. Indeed, the simultaneous presence of different genes - in a given evolutionary stage and with functions that are also partially correlated - would seem to define a heterogeneity that would appear to be the expression of a rapid and progressive evolution precisely because this evolution would have taken place in the diversification of all these genes. Furthermore, the presence of different genes coding for the function of cell division and related functions could reflect a progenotic status in LUCA, precisely because these functions might have originated from a single ancestral gene instead coding for a protein (or proteins) with multiple functions, and therefore an expression of a rapid and progressive evolution typical of the progenote. I also criticize other aspects of considerations made by Pende at al. (2021). The arguments presented here together with those existing in the literature make the hypothesis of a cellular LUCA favoured by Pende et al. (2021) unlikely.

Hoverflies (Diptera: Syrphidae) of the genus Microdon Meigen have larvae that live in ant nests where they are predatory on ant larvae. Reflecting the exceptional challenges of this very specialized lifestyle, Microdon eggs, larvae and puparia are highly distinctive in their morphology. Detailed descriptions of these immature stages is, however, lacking for all but a very few species, and much of this has been limited through the sole use of light microscopes. Here, using Scanning Electron Microscopy (SEM), we present detailed, comparative descriptions of the immature stages of three European Microdon species: M. analis, M. devius and M. myrmicae. Given that many adult Microdon species are very similar to each other in their outward appearance, we demonstrate that the morphology of their immature stages can improve our understanding of the phylogeny of the genus. We also discuss how particular adaptations of the immature morphology may allow their myrmecophilous life within ant nests. In this paper new diagnostic features are also presented to distinguish M. myrmicae from its sibling species M. mutabilis—the two are morphologically indistinguishable as adults.

Herein we revise several Recent Mediterranean species of the rissoid genus Alvania Risso, 1826: Alvania scabra (Philippi, 1844), Alvania sculptilis (Monterosato, 1877), Alvania sororcula Granata-Grillo, 1877, Alvania lucinae Oberling, 1970, Alvania josefoi Oliver & Templado, 2009 and Alvania scuderii Villari, 2017. They represent a rather homogeneous group of morphologically similar species, referred to as the Alvania scabra complex, which includes also some other species from the northeastern Atlantic. We designate a neotype for Rissoa scabra Philippi, 1844 and a lectotype for Rissoa oranica Pallary, 1900 to stabilize the use of the names. Alvania oranica (Pallary, 1900) is confirmed as a synonym of Alvania scabra (Philippi, 1844), and Alvania asperella (Granata-Grillo, 1877) is proposed as a synonym of Alvania sororcula (Granata-Grillo, 1877) [new synonymy]. Finally, we describe one new Mediterranean species: Alvania pizzinii Amati, Smriglio & Oliverio n. sp. from Levanzo Is., Sicily.

Environmental thermal conditions play a major role at all levels of biological organization; however, there is little information on noxious high temperature sensation crucial in behavioral thermoregulation and survival of small ectothermic animals such as insects. So far, a capability to unambiguously encode heat has been demonstrated only for the sensory triad of the spike bursting thermo- and two bimodal hygro-thermoreceptor neurons located in the antennal dome-shaped sensilla (DSS) in a carabid beetle. We used extracellular single sensillum recording in the range of 20–45°C to demonstrate that a similar sensory triad in the elaterid Agriotes obscurus also produces high temperature-induced bursty spike trains. Several parameters of the bursts are temperature dependent, allowing the neurons in a certain order to encode different, but partly overlapping ranges of heat up to lethal levels in a graded manner. ISI in a burst is the most useful parameter out of six. Our findings consider spike bursting as a general, fundamental quality of the classical sensory triad of antennal thermo- and hygro-thermoreceptor neurons widespread in many insect groups, being a flexible and reliable mode of coding unfavorably high temperatures. The possible involvement of spike bursting in behavioral thermoregulation of the beetles is discussed. By contrast, the mean firing rate of the neurons in regular and bursty spike trains combined does not carry useful thermal information at the high end of noxious heat.

Invasive alien species (IAS) are considered one of the largest drivers of biodiversity loss worldwide and the negative impacts of IAS can also affect human health and economy. More than 12,000 alien species occur in Europe. Terrestrial invertebrate species are the largest proportion of alien animal species and represents one of the most impacting groups in Europe. The most important European policy against alien species is the EU Regulation 1143/2014, which provides for the development of priority lists of IAS of relevant concern aimed to allow the optimization of intervention measures. Italian policy implemented the EU Regulation with the Legislative Decree No 230/2017 that provides the adoption of a list of IAS of national concern. Aim of this work is to present a preliminary prioritized list of alien terrestrial invertebrate species (ATIS) present in Italy, thus providing an useful tool to identify species to be included in the list of IAS of national concern. We defined criteria for assessing the species and ranking them in a prioritized list on the basis of the magnitude of their potential impact on wild native biodiversity. We identify 233 relevant ATIS, among the 1126 species included in the Italian Alien Terrestrial Invertebrate Database, on which the evaluation process started. After the evaluation process, 109 ATIS with considerable impacts on biodiversity were selected and prioritized. We ranked the species in four priority categories by matching their distribution in Italy with the magnitude of their possible impact on biodiversity.

Blister beetles owe their name to their ability to release cantharidin, a blistering terpene, the highest concentration of which is retained in male accessory glands. The anatomy and ultrastructure of the three pairs of male reproductive accessory glands and the glandular region of the two vasa deferentia of Meloe proscarabaeus were investigated using light, electron and ion beam microscopy. All of the mesodermal glands here analysed share a common structural organization with an outer muscular layer and an inner glandular epithelium facing a broad lumen in which the secretory products are released. Developed rough endoplasmic reticulum, Golgi systems, abundant mitochondria, numerous secretory vesicles and a microvillated apical membrane are commonly found in the cells of different glandular epithelia, suggesting that all accessory gland pairs as well as the vasa deferentia are involved in an active synthesis. Nevertheless, each pair of glands appears specialized in the production of a specific set of substances, as suggested by the peculiarities in cellular ultrastructure and by the different aspect of the secretions stored in their glandular lumen. The above cited features of male accessory glands of M. proscarabaeus are compared with those of other beetles and some hints on their potential role in producing and/or concentrating cantharidin are provided.

Many studies have highlighted how numerous bacteria provide their hosts essential nutrients or protection against pathogens, parasites and predators. Nevertheless, the role of symbiotic microorganisms in the interactions between social insects and their parasites is still poorly known. Microdon (Diptera, Syrphidae) is a peculiar fly genus whose larvae are able to successfully infiltrate ant colonies and feed upon the ant brood. Using high throughput 16S rRNA gene amplicon sequencing, we provide the first microbiome survey of Mi. myrmicae larvae and larvae and workers of its host, Myrmica scabrinodis, collected from two sites in England. We analyzed the microbiome of the external surface of the cuticle and the internal microbiome of the body separately. The results clearly show that the Mi. myrmicae microbiome significantly differs from that of its host, while no substantial dissimilarity was detected across the microbiome of ant workers and ant larvae. Microdon myrmicae microbiome varies across the two analyzed sites suggesting that bacteria communities of Mi. myrmicae are derived from the environment rather than by horizontal transmission between hosts and parasites. Families Streptococcaceae, Carnobacteriaceae and Rizhobiaceae are dominant in My. scabrinodis, and Spiroplasma is dominant in ant workers. Microbiome of Mi. myrmicae larvae is mainly characterized by the family Anaplasmataceae, with Wolbachia as predominant genus. Interestingly, we found Serratia within both Mi. myrmicae and Myrmica larvae. Bacteria of this genus are known to produce a family of pyrazines commonly involved in ant communication, which could play a role in Microdon/ant interaction.

Microbial symbionts enable many phytophagous insects to specialize on plant-based diets through a range of metabolic services. Pollen comprises one plant tissue consumed by such herbivores. While rich in lipids and protein, its nutrient content is often imbalanced and difficult-to-access due to a digestibly recalcitrant cell wall. Pollen quality can be further degraded by harmful allelochemicals. To identify microbes that may aid in palynivory, we performed cDNA-based 16S rRNA metabarcoding on three related pollen beetles (Nitidulidae: Meligethinae) exhibiting different dietary breadths: Brassicogethes aeneus, B. matronalis, and Meligethes atratus. Nine bacterial symbionts (i.e. 97% OTUs) exhibited high metabolic activity during active feeding. Subsequent PCR surveys revealed varying prevalence of those from three Rickettsialles genera - Lariskella, Rickettsia and Wolbachia - within beetle populations. Our findings lay the groundwork for future studies on the influence of phylogeny and diet on palynivorous insect microbiomes, and roles of symbionts in the use of challenging diets.

In the European Union between 2008 and 2017 there was a statistically significant decrease in the confirmed cases of salmonellosis. Nevertheless, it represents the first cause of food-borne outbreaks and the secondly most frequently reported zoonosis. Considering the pathogen diffusion and clinical consequences, monitoring the incidence of Salmonella spp. infections is a priority for public health. The study is aimed at describing the temporal trend of prevalence of Salmonella spp. positive tested and serotypes characteristics of the isolates by L’Aquila hospital laboratory microbiological analysis in 2011-2018, with a focus on the epidemic phase occurred from October 2013 to May 2014. The survey is a case series report, with a retrospective design. During the period considered, 220 infected carriers were identified, in particular 160 positive samples for Salmonella enterica serogroup B, so it was the main group causing salmonellosis in the area of L’Aquila both in the epidemic period and in the non-epidemic period. Moreover, the proportion of cases in early paediatric age (from one to five years) resulted statistically higher in the epidemic period than in the non-epidemic period. About the possible causes of the outbreak, the hypothesis of an environmental contamination seems to be plausible.

The myrmecophilous immature stages of hover fl ies of the genus Microdon Meigen, 1803 (Diptera, Syrphidae) are still poorly known and only about 15 species were previously incompletely described and/or illustrated using light microscopy based on occasional fi ndings mainly of pupae and third instar larvae. The exceptional fi nding of a large number of second and third instar larvae and pupae (159 specimens) of Microdon mutabilis (Linnaeus, 1758) inside the nest of a new host species, Formica cunicularia Latreille,1798, enabled us to rear them and obtain a great number of eggs and fi rst instar larvae. We fi lmed and described the feeding behaviour and locomotion of these highly derived slug-like larvae. Combining light, fl uorescence and scanning electron (SEM) microscopy, we describe in detail and illustrate the external features of all the immature stages of M. mutabilis (eggs, larvae and pupae). Covering the entire chorion of the egg is a peculiar microsculpture composed of volcano-like processes. The three larval instars strongly differ from each other, especially at the level of the shape of the body, the posterior spiracular tubercle and the cephaloskeleton. SEM microscopy was used to describe in detail the microsculpture, sensorial structures, spiracles and cephalic appendages of larvae and pupae. Fluorescence microscopy was used to reveal the exceptional presence of resilin in the external layer of the posterior spiracular tubercle in fi rst instar larvae. The possible functional signifi cance of these structures is discussed.

Sites along the Elizabeth River are contaminated with polycyclic aromatic hydrocarbons (PAHs) from historical creosote production and other industrial processes. Previous studies have demonstrated that Atlantic killifish collected from sites throughout the Elizabeth River display resistance to the teratogenic effects of PAH-exposure in a manner commensurate with sediment PAH concentrations. The current study characterized various chemical pollutants in sediment and investigated the effects of aqueous sediment extracts from sites along the Elizabeth River to the cardiac development of Atlantic killifish embryos from fish collected from an uncontaminated reference site. Embryonic cardiac deformities were more prevalent after exposure to extracts from sites with high PAH loads. However, activation of cytochrome P4501A, a gene up-regulated by PAH-induction of the aryl hydrocarbon receptor and measured using an in ovo EROD assay, did not consistently increase with PAH concentrations. This work further characterizes sediments in the Elizabeth River, as well as provides insight into the evolutionary pressures at each ER site.

LncRNAs play crucial roles in cellular processes and their regulatory effects in the adult brain and neural stem cells (NSCs) remain to be entirely characterized. We report that 10 lncRNAs (LincENC1, FABL, lincp21, HAUNT, PERIL, lincBRN1a, lincBRN1b, HOTTIP, TUG1 and FENDRR) are expressed during murine NSCs differentiation and interact with the RNA-binding protein ELAVL1/HuR. Furthermore, we characterize the function of two of the deregulated lncRNAs, lincBRN1a and lincBRN1b, during NSCs’ differentiation. Their inhibition leads to the induction of differentiation, with a concomitant decrease in stemness and an increase in neuronal markers, indicating that they exert key functions in neuronal cells differentiation. Furthermore, we describe here that HuR regulates their half-life, suggesting their synergic role in the differentiation process. We also identify six human homologs (PANTR1, TUG1, HOTTIP, TP53COR, ELDRR and FENDRR) of the mentioned 10 lncRNAs and we report their deregulation during human iPSCs differentiation into neurons. In conclusion, our results strongly indicate a key synergic role for lncRNAs and HuR in neuronal stem cells fate.

Plastics are ubiquitous anthropogenic contaminants that are a growing concern in aquatic environments. The ecological implications of macroplastics pollution are well documented, but less is known about nanoplastics. The current study investigates the potential adverse effects of nanoplastics, which likely contribute to the ecological burden of plastic pollution. To this end, we examined whether a dietary exposure of adult zebrafish (Danio rerio) to polystyrene nanoparticles (PS NPs) could lead to the transfer of nanoplastics to the offspring, and whether nanoplastics exposure affects zebrafish physiology. Specifically, adult female and male zebrafish (F0 generation) were exposed to PS NPs via diet for one week and bred to produce the F1 generation. Four F1 groups were generated: control (unexposed females and males), maternal (exposed females), paternal (exposed males), and co-parental (exposed males and females). Co-parental PS NP exposure did not significantly affect reproductive success. Assessment of tissues from F0 fish revealed that exposure to PS NPs significantly reduced glutathione reductase activity in brain, muscle, and testes, but did not affect mitochondrial function parameters in heart or gonads. Assessment of F1 embryos and larvae revealed that PS NPs were present in the yolk sac, gastrointestinal tract, liver, and pancreas of the maternally and co-parentally exposed F1 embryos/larvae. Bradycardia was also observed in embryos from maternal and co-parental exposure groups. In addition, the activity of glutathione reductase and the levels of thiols were reduced in F1 embryos/larvae from maternal and/or co-parental exposure groups. Mitochondrial function and locomotor activity were not affected in F1 larvae. This study demonstrates that (i) PS NPs are transferred from mothers to offspring, and (ii) exposure to PS NPs modifies the antioxidant system in adult tissues and F1 larvae. We conclude that PS NPs could bioaccumulate and be passed on to the offspring, but this does not lead to major physiological disturbances.

I analyse the origin of the genetic code in the light of the evolution of biological catalysts. I discuss the rudimentary forms that the genetic code assumed in the presence of a catalysis performed by ions or by low molecular weight molecules, such as nucleotide coenzymes. However, it is only with the advent of a mixed polymer made of RNA and peptides - covalently linked - that the genetic code took on a clearer form. Indeed, the first true form of coding appeared. Furthermore, interacting peptidated RNAs promoted an extremely rudimentary form of protein synthesis. This stage evolved into a stage in which proto-mRNAs guided interactions among peptidated RNAs aimed at the synthesis of peptidated RNAs having an active catalytic function. Finally, the invasion of aminoacylated proto-tRNAs with specific amino acids, coming from amino acid metabolism, and recognising only three bases on these proto-mRNAs with reading frames larger than three bases, would have triggered the birth of actual mRNAs, i.e. the origin of codons. All this would have linked the metabolism of amino acids to the origin of mRNAs and therefore to the origin of the organization of the genetic code, as maintained by the coevolution theory of the genetic code.

I analyzed the implications of the observation that the methyltransferases, Trm5 and TrmD, which perform the methylation of the 37th base (m1G37) in tRNAs of bacteria and archaea respectively, are not homologous proteins. The first implication is that these methyltransferases originated very late only when the fundamental lineages leading to bacteria and archaea had separated, otherwise the two methyltransferases would have been homologous enzymes, which they are not. The conclusion that Trm5 and TrmD originated only when the main lineages were defined would imply that at least some aspects of the translation, such as +1 frameshifting, were still in rapid and progressive evolution, that is, they were still originating. This would in itself imply a high rate of translation errors because the absence of m1G37 from tRNAs could have determined a high rate of +1 translational frameshifting in the reading of mRNAs, identifying this stage as that of a phase of the origin of the genetic code. Furthermore, the observation that the frameshifting mechanism was still in rapid and progressive evolution in such an advanced evolutionary stage would imply that other mechanisms concerning translation were still rapidly evolving simply because it would be very unique if only the frameshifting mechanism were the only one still originating. Importantly, the observation that in archaea m1G37 also acts as a determinant of the identity of the tRNACysGCA would imply in itself that some aspects of the origin of the genetic code were still originating, greatly strengthening the hypothesis that other aspects of the translation apparatus were still in rapid and progressive evolution. Then, all this would imply a status of progenote for LUCA and ancestors of archaea and bacteria because a high rate of translation errors would fall within the definition of progenote.

Organophosphate (OP) compounds comprise one of the most widely used classes of insecticides worldwide. OPs have been shown to have negative human health impacts, particularly developmental neurotoxicity. However, neurotoxic impacts in later adulthood and during the aging process are relatively uncharacterized. The present study examined diazinon (DZN), an OP, to determine the neurobehavioral consequences, in addition to mitochondrial dysfunction on a macroscale (whole organism basal respiration) and on a microscale (whole organ mitochondrial respiration), using zebrafish (ZF) as a model. One group of 14-month-old adult ZF were exposed acutely as adults (0.4, 1.25, and 4.0 μM) for five days and tested as adults, and another group was exposed developmentally 5–120 h post-fertilization (70, 210, and 700 nM) and tested at larval, adolescent, adult, and aging life stages. ZF exposed acutely as adults did not display many significant neurobehavioral impacts or mitochondrial dysfunction. Conversely, the embryonically exposed ZF showed altered behavioral functions at each stage of life which emerged and attenuated as fish transitioned from each developmental stage to the next. Mitochondrial oxygen consumptions measurement results for developmentally DZN exposed ZF showed significant increases in the low and middle dose groups in organs such as the brain and testes. Overall, there is an indication that early developmental exposure to DZN had continuing adverse neurobehavioral and cellular consequences throughout their lives well into adulthood and aging periods.

MYCN drives aggressive behavior and refractoriness to chemotherapy, in several tumors. Since MYCN inactivation in clinical settings is not achievable, alternative vulnerabilities of MYCN-driven tumors need to be explored to identify more effective and less toxic therapies. We previously demonstrated that PARP inhibitors enhance MYCN-induced replication stress and promote mitotic catastrophe, counteracted by CHK1. Here, we showed that PARP and CHK1 inhibitors synergized to induce death in neuroblastoma cells and in primary cultures of SHH-dependent medulloblastoma, their combination being more effective in MYCN amplified and MYCN overexpressing cells compared to MYCN non-amplified cells. Although the MYCN amplified IMR-32 cell line carrying the p.Val2716Ala ATM mutation showed the highest sensitivity to the drug combination, this was not related to ATM status, as indicated by CRISPR/Cas9-based correction of the mutation. Suboptimal doses of the CHK1 inhibitor MK-8776 plus the PARP inhibitor olaparib led to a MYCN-dependent accumulation of DNA damage and cell death in vitro and significantly reduced the growth of four in vivo models of MYCN-driven tumors, without major toxicities. Our data highlight the combination of PARP and CHK1 inhibitors as a new potential chemo-free strategy to treat MYCN-driven tumors, which might be promptly translated into clinical trials.

Induced pluripotent stem cells (iPSCs) hold great promise for developing personalized regenerative medicine, however characterization of their biological features is still incomplete. Moreover, changes occurring in long-term cultured iPSCs have been reported, suggesting these as a model of cellular aging. For this reason, we addressed the ultrastructural characterization of iPSCs, with a focus on possible time-dependent changes, involving specific cell compartments. To this aim, we comparatively analysed cultures at different timepoints, by an innovative electron microscopic technology (FIB/SEM). We observed progressive loss of cell-to-cell contacts, associated with increased occurrence of exosomes. Mitochondria gradually increased, while acquiring an elongated shape, with well-developed cristae. Such mitochondrial maturation was accompanied by their turnover, as assessed by the presence of autophagomes (undetectable in young iPSCs), some containing recognizable mitochondria. This finding was especially frequent in middle-aged iPSCs, while being occasional in aged cells, suggesting early autophagic activation followed by a decreased efficiency of the process with culturing time. Accordingly, confocal microscopy showed age-dependent alterations to the expression and distribution of autophagic markers. Interestingly, responsivity to rapamycin, highest in young iPSCs, was almost lost in aged cells. Overall, our results strongly support long-term cultured iPSCs as a model for studying relevant aspects of cellular senescence, involving intercellular communication, energy metabolism, and autophagy.

Members of the globally distributed bivalve family Ostreidae (oysters) have a significant role in marine ecosystems and include species of high economic importance. In this work, we report the occurrence of digenean parasites of the families Bucephalidae (Prosorhynchoides sp.) and Monorchiidae (Postmonorchis sp.) in Mediterranean native populations of Ostrea edulis (but not in the introduced Magallana gigas). Molecular detection was based on DNA sequencing of the ribosomal intergenic spacer 2 (ITS2) marker. The importance of detecting the presence of overlooked digenean parasites in Mediterranean oysters is discussed.

Despite thermosensation being crucial in effective thermoregulation behaviour, it is poorly studied in insects. Very little is known about encoding of noxious high temperatures by peripheral thermoreceptor neurons. In carabids, thermo- and hygrosensitive neurons innervate antennal dome-shaped sensilla (DSS). In this study, we demonstrate that several essential fine structural features of dendritic outer segments of the sensory neurons in the DSS and the classical model of insect thermo- and hygrosensitive sensilla differ fundamentally. Here, we show that spike bursts produced by the bimodal dry neurons in the antennal DSS may contribute to the sensation of noxious heat in P. oblongopunctatus. Our electrophysiological experiments showed that, at temperatures above 25 °C, these neurons switch from humidity-dependent regular spiking to temperature-dependent spike bursting. Five out of seven measured parameters of the bursty spike trains, the percentage of bursty dry neurons, the CV of ISIs in a spike train, the percentage of bursty spikes, the number of spikes in a burst and the ISIs in a burst, are unambiguously dependent on temperature and thus may precisely encode both noxious high steady temperatures up to 45 °C as well as rapid step-changes in it. The cold neuron starts to produce temperature-dependent spike bursts at temperatures above 30-35 °C. Thus, the two neurons encode different but largely overlapping ranges in noxious heat. The extent of dendritic branching and lamellation of the neurons largely varies in different DSS, which might be the structural basis for their variation in threshold temperatures for spike bursting.

Extensive molecular characterization of human colorectal cancer (CRC) via Next Generation Sequencing (NGS) indicated that genetic or epigenetic dysregulation of a relevant, but limited, number of molecular pathways typically occurs in this tumor. The molecular picture of the disease is significantly complicated by the frequent occurrence of individually rare genetic aberrations, which expand tumor heterogeneity. Inter- and intratumor molecular heterogeneity is very likely responsible for the remarkable individual variability in the response to conventional and target-driven first-line therapies, in metastatic CRC (mCRC) patients, whose median overall survival remains unsatisfactory. Implementation of an extensive molecular characterization of mCRC in the clinical routine does not yet appear feasible on a large scale, while multigene panel sequencing of most commonly mutated oncogene/oncosuppressor hotspots is more easily achievable. Here, we report that clinical multigene panel sequencing performed for anti-EGFR therapy predictive purposes in 639 formalin-fixed paraffin-embedded (FFPE) mCRC specimens revealed previously unknown pairwise mutation associations and a high proportion of cases carrying actionable gene mutations. Most importantly, a simple principal component analysis directed the delineation of a new molecular stratification of mCRC patients in eight groups characterized by non-random, specific mutational association patterns (MAPs), aggregating samples with similar biology. These data were validated on a The Cancer Genome Atlas (TCGA) CRC dataset. The proposed stratification may provide great opportunities to direct more informed therapeutic decisions in the majority of mCRC cases.

Various organisms, especially arthropods, are able to live as parasites in ant nests and to prey upon ant broods without eliciting any aggressive behaviour in the hosts. Understanding how these intruders are able to break the ants' communication codes in their favour represents a challenging and intriguing evolutionary question. We studied the chemical strategies of three European hoverfly species, Microdon mutabilis (parasitic on Formica cunicularia), M. analis (parasitic on Lasius emarginatus) and M. devius (parasitic on L. distinguendus). The peculiar slug-like larvae of these three species live inside ant nests feeding upon their broods. Gas chromatography-mass spectrometry analyses show that: 1) these parasites mimic the host brood rather than the ant workers, although each differs distinctly in the extent of chemical mimicry; 2) isolation experiments indicate that after 14days the responsible cuticular hydrocarbons (CHCs) are not passively acquired but synthesized by the fly larvae. Additionally, Microdon larvae show an array of protective structural features, such as a thick and multi-layered cuticle, retractable head, dome-shaped tergum and a flat and strongly adhesive "foot" (sternum). This combination of protective chemical and structural features represents a successful key innovation by Microdon species, and one that may facilitate host switching. The results of a preliminary adoption analysis confirm that Microdon larvae of at least some species can readily be accepted by different species of ants.

Stevenia gilasiani Ziegler, Gisondi & Cerretti, sp. nov. from western Iran is described and illustrated. Cladistic arguments are provided in order to give an explicit genus level affiliation for the new species. Stevenia gilasiani sp. nov. is the first rhinophorid characterized by male ventral sexual-patches on abdominal tergites 3, 4 and 5 and sternites 3 and 4, and is the only Stevenia characterized by two postpronotal setae instead of usual three. A comparison with all congeners is made.

The carabid subfamily Paussinae contains many species known to be obligately associated with ants during at least one stage of their life history. Myrmecophilous larvae have been documented for members of the tribe Paussini as well as several genera in the tribes Ozaenini, including Physea and Eustra. Here we describe the first instar larva of Ozaena lemoulti, and find it to be the most highly modified ozaenine larva that we have examined to date. Many structures of the larva suggest that it is a myrmecophile. Unlike all other described ozaenine larvae, which live in burrows that they construct and seal with their terminal disk, the completely unique larval morphology suggests Ozaena has adapted to living without the protection of a burrow and therefore must have a completely different feeding strategy than the typical ambush strategy of burrow dwelling larvae. We hypothesize that Ozaena larvae live in association with ants and use their long legs for running within the nest, and modifications of the mouthparts suggest the larva feeds on soft lightly sclerotized prey, such as ant brood. Our findings support an earlier hypothesis that Ozaena is mymecophilous during the adult stage. Comparisons of the functional anatomy of the eggs, larvae and adult between Ozaena lemoulti and the closely related, non-myrmecophilous general arthropod predator, Goniotropis kuntzeni, provide complementary, yet independent, evidence suggestive of this shift in lifestyle. We also examine and molecularly identify gut contents, providing direct evidence that adult Ozaena exclusively eat Camponotus ants. We conclude that Ozaena represents an independent shift to adopting a life of myrmecophily among beetles classified within the carabid subfamily Paussinae and document the morphological changes at each life history stage associated with the shift to a nest parasite lifestyle.

It has been firmly observed that replicative DNA polymerases of bacteria, archaea and eukaryotes are not homologous proteins. This lack of homology in the replication apparatus among the domains of life is not only compatible with but would seem to imply the view that the emergence of DNA occurred in the fundamental cellular lineages. In consequence, this diversity of DNA polymerase would go back to the level of ancestors of the domains of life and to the evolutionary time in which the DNA emerged. Therefore, the presumed evolutionary stage linked to the RNA- > DNA transition would have occurred only at the level of ancestors of the main lineages of the tree of life. Thus, the high noise associated with this major evolutionary transition and the impossibility for a cellular stage to generate different fundamental genetically profound traits - such as the different replication apparatuses of bacteria, archaea and eukaryotes - would imply not only that the last universal common ancestor (LUCA) was a progenote but that the ancestors of the domains of life were also at this evolutionary stage. So, I criticize the hypotheses which want, instead, that completely different cells - such as, bacteria and archaea - could have originated from a cellular LUCA.

Cerebrospinal fluid contacting neurons (CSF-cNs) represent a specific class of neurons located in close vicinity of brain ventricles and central canal. In contrast with knowledge gained from other vertebrate species, we found that vast majority of CSF-cNs in the spinal cord of C57Bl/6N mice is located in ectopic distal ventral position. However, we found that small number of ectopic CSF-cNs is present also in spinal cord of other investigated experimental mice strains (C57Bl/6J, Balb/C) and mammalian species (Wistar rats, New Zealand White rabbits). Similarly, as the proximal populations, ectopic CSF-cNs retain PKD2L1-immunoreactivity and synaptic contacts with other neurons. On the other side, they show rather multipolar morphology lacking thick dendrite contacting central canal lumen. Ectopic CSF-cNs in the spinal cord of C57Bl/6N mice emerge during whole period devoted to production of CSF-cNs and reach their ventral destinations during first postnatal weeks. In order to identify major gene, whose impairment could trigger translocation of CSF-cNs outside the central canal area, we took advantage of close consanguinity of C57Bl/6J substrain with normal CSF-cN distribution and C57Bl/6N substrain with majority of CSF-cNs in ectopic position. Employing in silico analyses, we ranked polymorphisms in C57Bl/6N substrain and selected genes Crb1, Cyfip2, Adamts12, Plk1, and Herpud2 as the most probable candidates, whose product dysfunction might be responsible for the ectopic distribution of CSF-cNs. Furthermore, segregation analysis of F2 progeny of parental C57Bl/6N and Balb/C mice revealed that polymorphic loci of Crb1 and Cyfip2 underlie the ectopic position of CSF-cNs in the spinal cord of C57Bl/6N mice.