Development of pH-Sensitive Magnetoliposomes Containing Shape Anisotropic Nanoparticles for Potential Application in Combined Cancer Therapy.

Late diagnosis and systemic toxicity associated with conventional treatments make oncological therapy significantly difficult. In this context, nanomedicine emerges as a new approach in the prevention, diagnosis and treatment of cancer. In this work, pH-sensitive solid magnetoliposomes (SMLs) were developed for controlled release of the chemotherapeutic drug doxorubicin (DOX). Shape anisotropic magnetic nanoparticles of magnesium ferrite with partial substitution by calcium (Mg0.75Ca0.25Fe2O4) were synthesized, with and without calcination, and their structural, morphological and magnetic properties were investigated. Their superparamagnetic properties were evaluated and heating capabilities proven, either by exposure to an alternating magnetic field (AMF) (magnetic hyperthermia) or by irradiation with near-infrared (NIR) light (photothermia). The Mg0.75Ca0.25Fe2O4 calcined nanoparticles were selected to integrate the SMLs, surrounded by a lipid bilayer of DOPE:Ch:CHEMS (45:45:10). DOX was encapsulated in the nanosystems with an efficiency above 98%. DOX release assays showed a much more efficient release of the drug at pH = 5 compared to the release kinetics at physiological pH. By subjecting tumor cells to DOX-loaded SMLs, cell viability was significantly reduced, confirming that they can release the encapsulated drug. These results point to the development of efficient pH-sensitive nanocarriers, suitable for a synergistic action in cancer therapy with magnetic targeting, stimulus-controlled drug delivery and dual hyperthermia (magnetic and plasmonic) therapy.

Genomic analysis of a multidrug methicillin-resistant staphylococcus epidermidis recovered from the urine of a guinea pig (Cavia porcellus) with suspected pyelonephritis.

Staphylococcus epidermidis is an emerging pathogen causing infant pyelonephritis. There is a lack of genomic data on Staphylococcus epidermidis as the etiology of pyelonephritis and its resistant determinants. In this study, we have conducted a genomic and microbiologic investigation of an S. epidermidis recovered from the urine of a guinea pig with suspected pyelonephritis in Brazil. The genome was sequenced using the Illumina MiSeq platform and de novo assembled using SPades. Resistome, virulome, and plasmidome were in silico predicted using bioinformatics tools. Data analysis revealed that S. epidermidis USP-LZB-G06 belonged to sequence type ST332. Strikingly, a broad resistome (antibiotics, hazardous heavy metals, and biocides) was predicted, including the presence of the clinically relevant mecA, blaZ, and qacA efflux pump genes. SNP-based analysis revealed that strain USP-LZB-G06 was clustered along mecA positive S. epidermidis strains of ST332 isolated between 2008 and 2016 from humans in Australia and the United States of America. Our results indicate that the detection of this microorganism should be considered as a urinary tract infection agent in exotic pets, particularly guinea pigs. In addition, there is an urgent need to update veterinarians regarding the detection and therapeutic management of these microorganisms.

Magnetoliposomes Containing Multicore Nanoparticles and a New Antitumor Thienopyridine Compound with Potential Application in Chemo/Thermotherapy.

Multicore magnetic nanoparticles of manganese ferrite were prepared using carboxymethyl dextran as an agglutinating compound or by an innovative method using melamine as a cross-coupling agent. The nanoparticles prepared using melamine exhibited a flower-shape structure, a saturation magnetization of 6.16 emu/g and good capabilities for magnetic hyperthermia, with a specific absorption rate (SAR) of 0.14 W/g. Magnetoliposome-like structures containing the multicore nanoparticles were prepared, and their bilayer structure was confirmed by FRET (Förster Resonance Energy Transfer) assays. The nanosystems exhibited sizes in the range of 250-400 nm and a low polydispersity index. A new antitumor thienopyridine derivative, 7-[4-(pyridin-2-yl)-1H-1,2,3-triazol-1-yl]thieno[3,2-b]pyridine, active against HeLa (cervical carcinoma), MCF-7 (breast adenocarcinoma), NCI-H460 (non-small-cell lung carcinoma) and HepG2 (hepatocellular carcinoma) cell lines, was loaded in these nanocarriers, obtaining a high encapsulation efficiency of 98 ± 2.6%. The results indicate that the new magnetoliposomes can be suitable for dual cancer therapy (combined magnetic hyperthermia and chemotherapy).

Four New Species of Harringtonia: Unravelling the Laurel Wilt Fungal Genus.

Symbiosis between beetles and fungi arose multiple times during the evolution of both organisms. Some of the most biologically diverse and economically important are mutualisms in which the beetles cultivate and feed on fungi. Among these are bark beetles and Harringtonia, a fungal genus that produces Raffaelea-like asexual morph and hosts the causal agent of laurel wilt, H. lauricola (formerly Raffaelea lauricola). In this study, we propose four new species of Harringtonia associated with beetles from Belize and Florida (USA). We hope to contribute towards a more robust and inclusive phylogenetic framework for future studies on these beetle-fungi relationships and their potential impact in crops and forests worldwide.

Development of Thermo- and pH-Sensitive Liposomal Magnetic Carriers for New Potential Antitumor Thienopyridine Derivatives.

The development of stimuli-sensitive drug delivery systems is a very attractive area of current research in cancer therapy. The deep knowledge on the microenvironment of tumors has supported the progress of nanosystems' ability for controlled and local fusion as well as drug release. Temperature and pH are two of the most promising triggers in the development of sensitive formulations to improve the efficacy of anticancer agents. Herein, magnetic liposomes with fusogenic sensitivity to pH and temperature were developed aiming at dual cancer therapy (by chemotherapy and magnetic hyperthermia). Magnetic nanoparticles of mixed calcium/manganese ferrite were synthesized by co-precipitation with citrate and by sol-gel method, and characterized by X-ray diffraction (XRD), scanning electron microscopy in transmission mode (STEM), and superconducting quantum interference device (SQUID). The citrate-stabilized nanoparticles showed a small-sized population (around 8 nm, determined by XRD) and suitable magnetic properties, with a low coercivity and high saturation magnetization (~54 emu/g). The nanoparticles were incorporated into liposomes of dipalmitoylphosphatidylcholine/cholesteryl hemisuccinate (DPPC:CHEMS) and of the same components with a PEGylated lipid (DPPC:CHEMS:DSPE-PEG), resulting in magnetoliposomes with sizes around 100 nm. Dynamic light scattering (DLS) and electrophoretic light scattering (ELS) measurements were performed to investigate the pH-sensitivity of the magnetoliposomes' fusogenic ability. Two new antitumor thienopyridine derivatives were efficiently encapsulated in the magnetic liposomes and the drug delivery capability of the loaded nanosystems was evaluated, under different pH and temperature conditions.

Changes in Intra-to-Extra-Cellular Water Ratio and Bioelectrical Parameters from Day-Before to Day-Of Competition in Bodybuilders: A Pilot Study.

The present study analyzed the effects from day-before to day-of bodybuilding competition on intracellular water (ICW), extracellular water (ECW), total body water (TBW), and bioimpedance analysis (BIA) parameters (resistance, R; reactance, Xc; and derived scores) in bodybuilding athletes. We assessed anthropometry and BIA (foot-to-hand; tetrapolar; 50 kHz) in 11 male bodybuilders (29 ± 4 year-old; 81 ± 8 kg; 172 ± 7 cm; 27 ± 2 kg/m2) both on the pre-competition day and on the contest day. Results revealed significant increases in ICW (31.6 ± 2.9 to 33.1 ± 2.8 L), with concomitant decreases in ECW (19.8 ± 1.8 to 17.2 ± 1.4 L) and TBW (51.4 ± 4.6 to 50.3 ± 4.2 L) from the day-before competition to contest day, which resulted in relatively large increases in the ICW/ECW ratio (1.60 ± 0.03 to 1.92 ± 0.01 L). Moreover, significant increases in R (391 ± 34 to 413 ± 33 ohm), Xc (64 ± 7 to 70 ± 6 ohm), and phase angle (9.3 ± 0.6 to 9.6 ± 0.7 degree) were observed between time periods. The phase angle scores reported on show-day of 9.6 and 11.2 appear to be the highest group mean and individual values observed in the literature to date. In conclusion, the strategies carried out on the final day of peak-week bodybuilding preparation lead to changes in BIA parameters and body water, with fluids shifting from the extra- to the intracellular compartment.

Diversity and Evolution of Entomocorticium (Russulales, Peniophoraceae), a Genus of Bark Beetle Mutualists Derived from Free-Living, Wood Rotting Peniophora.

Symbiosis between insects and fungi arose multiple times during the evolution of both groups, and some of the most biologically diverse and economically important are mutualisms in which the insects cultivate and feed on fungi. Among these are bark beetles, whose ascomycetous cultivars are better known and studied than their frequently-overlooked and poorly understood basidiomycetous partners. In this study, we propose five new species of Entomocorticium, fungal mutualists in the Russulales (Basidiomycota) that are mutualistic symbionts of scolytine beetles. We have isolated these fungi from the beetle mycangia, which are structures adapted for the selective storage and transportation of fungal mutualists. Herein, we present the most complete phylogeny of the closely related genera Entomocorticium and Peniophora and provide insights into how an insect-associated taxon (Entomocorticium) evolved from within a wood-decaying, wind-dispersed lineage (Peniophora). Our results indicate that following a transition from angiosperms to gymnosperms, fungal domestication by beetles facilitated the evolution and diversification of Entomocorticium. We additionally propose four new species: Entomocorticium fibulatum Araújo, Li & Hulcr, sp. nov.; E. belizense Araújo, Li & Hulcr, sp. nov.; E. perryae Araújo, Li & Hulcr, sp. nov.; and E. macrovesiculatum Araújo, Li, Six & Hulcr, sp. nov. Our findings highlight the fact that insect-fungi associations remain an understudied field and that these associations harbor a large reservoir of novel fungal species.

Magnetoliposomes Based on Magnetic/Plasmonic Nanoparticles Loaded with Tricyclic Lactones for Combined Cancer Therapy.

Liposome-like nanoarchitectures containing manganese ferrite nanoparticles covered or decorated with gold were developed for application in dual cancer therapy, combining chemotherapy and photothermia. The magnetic/plasmonic nanoparticles were characterized using XRD, UV/Visible absorption, HR-TEM, and SQUID, exhibiting superparamagnetic behavior at room temperature. The average size of the gold-decorated nanoparticles was 26.7 nm for MnFe2O4 with 5-7 nm gold nanospheres. The average size of the core/shell nanoparticles was 28.8 nm for the magnetic core and around 4 nm for the gold shell. Two new potential antitumor fluorescent drugs, tricyclic lactones derivatives of thienopyridine, were loaded in these nanosystems with very high encapsulation efficiencies (higher than 98%). Assays in human tumor cell lines demonstrate that the nanocarriers do not release the antitumor compounds in the absence of irradiation. Moreover, the nanosystems do not cause any effect on the growth of primary (non-tumor) cells (with or without irradiation). The drug-loaded systems containing the core/shell magnetic/plasmonic nanoparticles efficiently inhibit the growth of tumor cells when irradiated with red light, making them suitable for a triggered release promoted by irradiation.

Versatile Seebeck and electrical resistivity measurement setup for thin films.

A custom setup for Seebeck coefficient and electrical resistivity measurements of thin films as a function of temperature in the range of 10-300 K was developed. The Seebeck coefficient is measured using a two-probe arrangement and using either a dynamical or steady/quasi-steady differential method. The temperature differences (ΔTs) for these measurements across the samples are achieved by using resistive heaters embedded in two copper blocks. The sample is screwed to these blocks and is in pressured contact with the measurement probes. The electrical resistivity is measured with a two-probe arrangement. To verify the reliability of the developed setup, measurement tests were performed on commercial niobium foil and a specular spin valve previously studied, having obtained a great accordance (within ∼3%) between this setup's experimental results and the reference measurements.

Magnetic reversal modes in cylindrical nanostructures: from disks to wires.

Cylindrical magnetic nanowires are key elements of fast-recording and high-density 3D-storage devices. The accurate tuning of the magnetization processes at the nanoscale is crucial for the development of future nano-devices. Here, we analyzed the magnetization of Ni nanostructures with 15-100 nm in diameter and 12-230 nm in length and compared our results with experimental data for periodic arrays. Our modelling led to a phase diagram of the reversal modes where the presence of a critical diameter (d ≈ 30 nm) triggered the type of domain wall (DW) formed (transverse or vortex); while a critical length (L ≈ 100 nm) determined the number of DWs nucleated. Moreover, vortex-DWs originated from 3D skyrmion tubes, reported as one of the best configurations for storage devices. By increasing the diameter and aspect-ratio of nanowires with L > 100 nm, three reversal modes were observed: simultaneous propagation of two vortex-DWs; propagation of one vortex-DW; or spiral rotation of both DWs through "corkscrew" mechanism. Only for very low aspect-ratios (nanodisks), no skyrmion tubes were observed and reversal occurred by spiral rotation of one vortex-DW. The broad range of nanostructures studied allowed the creation of a complete phase diagram, highly important for future choice of nanoscaled dimensions in the development of novel nano-devices.

Group Theory Analysis to Study Phase Transitions of Quasi-2D Sr3Hf2O7.

We present an ab-initio study performed in the framework of density functional theory, group-subgroup symmetry analysis and lattice dynamics, to probe the octahedral distortions, which occur during the structural phase transitions of the quasi-2D layered perovskite Sr3Hf2O7 compound. Such a system is characterized by a high-temperature I4/mmm centrosymmetric structure and a ground-state Cmc21 ferroelectric phase. We have probed potential candidate polymorphs that may form the I4/mmm → Cmc21 transition pathways, namely Fmm2, Ccce, Cmca and Cmcm. We found that the band gap widths increase as the symmetry decreases, with the ground-state structure presenting the largest gap width (∼5.95 eV). By probing the Partial Density of States, we observe a direct relation regarding the tilts and rotations of the oxygen perovskite cages as the transition occurs; these show large variations mostly of the O p-states which contribute mostly to the valence band maximum. Moreover, by analyzing the hyperfine parameters, namely the Electric Field Gradients and asymmetric parameters, we observe variations as the transition occurs, from which it is possible to identify the most plausible intermediate phases. We have also computed the macroscopic polarization and confirm that the Cmc21 phase is ferroelectric with a value of spontaneous polarization of 0.0478 C/m2. The ferroelectricity of the ground-state Cmc21 system arises due to a second order parameter related to the coupling of the rotation and tilts of the O perovskite cages together with the Sr displacements.

Three-Dimensional (3D) Microporous Iron Silicate with an Imandrite Type of Structure.

Small-pore iron silicate MS-1 (Minho-Sofia, solid number 1) with a 3D porous system, an analogue of the rare mineral imandrite, has been synthesized and characterized. This material is the lowest framework density iron silicate, one of the most siliceous (Si/Fe = 6) iron silicates, the first iron cyclosilicate achieved at hydrothermal conditions, and the only synthetic iron-based member of the lovozerite mineral group.

Ophiocordyceps salganeicola, a parasite of social cockroaches in Japan and insights into the evolution of other closely-related Blattodea-associated lineages.

The entomopathogenic genus Ophiocordyceps includes a highly diverse group of fungal species, predominantly parasitizing insects in the orders Coleoptera, Hemiptera, Hymenoptera and Lepidoptera. However, other insect orders are also parasitized by these fungi, for example the Blattodea (termites and cockroaches). Despite their ubiquity in nearly all environments insects occur, blattodeans are rarely found infected by filamentous fungi and thus, their ecology and evolutionary history remain obscure. In this study, we propose a new species of Ophiocordyceps infecting the social cockroaches Salganea esakii and S. taiwanensis, based on 16 years of collections and field observations in Japan, especially in the Ryukyu Archipelago. We found a high degree of genetic similarity between specimens from different islands, infecting these two Salganea species and that this relationship is ancient, likely not originating from a recent host jump. Furthermore, we found that Ophiocordyceps lineages infecting cockroaches evolved around the same time, at least twice, one from beetles and the other from termites. We have also investigated the evolutionary relationships between Ophiocordyceps and termites and present the phylogenetic placement of O. cf. blattae. Our analyses also show that O. sinensis could have originated from an ancestor infecting termite, instead of beetle larvae as previously proposed.

Esteya floridanum sp. nov.: An Ophiostomatalean Nematophagous Fungus and Its Potential to Control the Pine Wood Nematode.

The nematophagous fungal genus Esteya is reported as a natural enemy of the pine wood nematode Bursaphelenchus xylophilus, which causes pine wilt in Asia and Europe. During a survey of fungi associated with ambrosia beetles in Florida, an undescribed Esteya species was found. A phylogenetic analysis based on nuclear large subunit and ß-tubulin DNA sequences supported this isolate as a new species, E. floridanum. Morphological and phylogenetic characteristics and a species description are provided here. The fungus was observed to kill the pine wood nematode in vitro. To evaluate the ability of E. floridanum to protect trees against the pine wood nematode in vivo, the effect of prophylactic inoculation was tested on Pinus koraiensis and Larix olgensis in Liaoning, China. The results suggest that the fungus is not a plant pathogen and that it delays wilt and postpones death of two conifer trees. This presents a potential new avenue for research on biocontrol of pine wilt disease and stresses the value of research on pest organisms in their native regions.

Moniliophthora perniciosa, the mushroom causing witches' broom disease of cacao: Insights into its taxonomy, ecology and host range in Brazil.

Witches' broom caused by Moniliophthora perniciosa is the main disease of cacao (Theobroma cacao) in Brazil. The fungus is known to occur on other host families and these populations have been addressed in the literature as biotypes: C (Malvaceae); H (Malpighiaceae); L (Bignoniaceae) and S (Solanaceae). No complete elucidation of the phylogenetic relationships of isolates obtained from this disparate host range appears in the literature. One member of H (ex Heteropterys acutifolia) has been described as a distinct species. But should other biotypes be also recognized as distinct taxa? In the present study, a survey yielding 24 isolates of M. perniciosa from ten hosts and covering a wide range of geographic regions in Brazil was undertaken. These isolates were compared with those from T. cacao using three DNA regions for the phylogenetic analyses: ITS, LSU and RPB1. Morphology was also examined. All isolates in this study were found to belong to M. perniciosa, including the population from H. acutifolia, formerly treated as Moniliophthora brasiliensis but reduced here to a synonym of M. perniciosa. This species ranged from pathogenic to a previously unreported occurrence as a non-pathogenic endophyte in the Atlantic rainforest tree Allophylus edulis (Sapindaceae). M. perniciosa was recorded on a range of solanaceous hosts (16 species) over a wide variety of ecosystems. The ecological and evolutionary significance of these novel findings are discussed.

A new zombie ant behavior unraveled: Aggregating on tree trunks.

Hosts can be manipulated by parasites to move to locations advantageous for onward transmission. To investigate the role of behavioral manipulation in creating transmission hotspots, we studied the distribution of zombie turtle ants in the Amazon rainforest. The turtle ant Cephalotes atratus nests and mostly forages in the canopy, but is found at the base of trees when infected with the zombie ant fungus Ophiocordyceps kniphofioides. We found 626 infected cadavers on 14.8% of 162 trees sampled. Cadavers were highly aggregated on the surface of the trees, explained by behavioral observations indicating infected ants as slightly attracted to zombie ant cadavers on a tree. From 1,726 h of camera footage, we recorded the removal of three zombie ant cadavers by live ants. The number of removals compared to the density of infected individuals indicates the base of a tree as an escape from the evolved ability of social insects to recognize and treat disease inside the nest, allowing the parasite to continuously remain in the environment.

Ophiocordyceps desmidiospora, a basal lineage within the "Zombie-Ant Fungi" clade.

The genus Ophiocordyceps contains the most diverse assemblage of fungi that attack ants worldwide and are remarkably well adapted to the specific ecologies of their hosts. Desmidiospora myrmecophila Thaxt. is closely related to other ant-pathogenic species within Ophiocordyceps, possibly specific to queens, but the sheer infrequency of encounters and previously unsuccessful attempts to culture this fungus has precluded any meaningful assessment until now. A new record of Desmidiospora myrmecophila from Louisiana was found infecting a foundress Camponotus pennsylvanicus queen, the same host species favored by the more common and ubiquitous ant-pathogenic Ophiocordyceps unilateralis clade found in the same geographic locality. To evaluate a long-held assumption that these fungi represent synanamorphs of a single species, we sampled our Desmidiospora specimen along with the local O. unilateralis population for molecular comparison. We are able to present for the first time the in vitro characteristics and morphology of Desmidiospora myrmecophila, as well as a phylogenetic context for this fungus based on combined molecular analysis of representative members of the Ophiocordycipitaceae. Our results place the Desmidiospora myrmecophila lineage within the genus Ophiocordyceps, with a basal affiliation to the Ophiocordyceps unilateralis core clade; thus, in accordance to the "One Fungus-One Name" (1F1N) rule, we propose a new synonym to suppress Desmidiospora in protection of Ophiocordyceps, i.e., O. desmidiospora. These results further implicate this species as an important and quintessential example of cryptic diversity among an already taxonomically diverse and ecologically important group of fungi.

Tailoring the Anodic Hafnium Oxide Morphology Using Different Organic Solvent Electrolytes.

Highly ordered anodic hafnium oxide (AHO) nanoporous or nanotubes were synthesized by electrochemical anodization of Hf foils. The growth of self-ordered AHO was investigated by optimizing a key electrochemical anodization parameter, the solvent-based electrolyte using: Ethylene glycol, dimethyl sulfoxide, formamide and N-methylformamide organic solvents. The electrolyte solvent is here shown to highly affect the morphological properties of the AHO, namely the self-ordering, growth rate and length. As a result, AHO nanoporous and nanotubes arrays were obtained, as well as other different shapes and morphologies, such as nanoneedles, nanoflakes and nanowires-agglomerations. The intrinsic chemical-physical properties of the electrolyte solvents (solvent type, dielectric constant and viscosity) are at the base of the properties that mainly affect the AHO morphology shape, growth rate, final thickness and porosity, for the same anodization voltage and time. We found that the interplay between the dielectric and viscosity constants of the solvent electrolyte is able to tailor the anodic oxide growth from continuous-to-nanoporous-to-nanotubes.

Zombie-Ant Fungi Emerged from Non-manipulating, Beetle-Infecting Ancestors.

The manipulation of animal behavior by parasitic organisms is one of the most complex adaptations to have arisen via natural selection. Among the most impressive examples of behavioral manipulation are the zombie-ant fungi [1]. In this association, ants are controlled to leave the colony and perform a stereotyped death grip behavior, where they bite onto vegetation over foraging trails, before being killed for the post mortem fungal growth. Manipulation functions to provide a platform outside the nest, from which fungal parasites actively shoot out spores, targeting foraging ants because within colony transmission is prevented by strong social immunity exhibited by social insect societies [2, 3]. It is not clear how such complex examples of host manipulation arose. To address this, we performed a broad-scale phylogenetic reconstruction of the order Hypocreales, to which the zombie-ant fungi, Ophiocordyceps, belong. In order to understand the patterns of host association and host switching along the evolution of Ophiocordyceps, we performed ancestral character state reconstruction analysis. We found that zombie-ant fungi likely arose from an ancestor that infected beetle larvae residing in soil or decaying wood, similar to extant beetle-infecting Ophiocordyceps species. Surprisingly, the jump led to an extensive species radiation observed after the development of behavioral manipulation. We suggest that the jump from solitary beetle larva to ants within a colony exposed the fungus to the robust social immunity of ant societies.