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We have sputter deposited NiO–Co bilayers on MgO (001) substrates. NiO grows epitaxially on MgO at 800 °C and subsequently room temperature deposited Co films have a fcc crystal structure in epitaxy with the oxide underlayer. Bias fields higher than 700 Oe at 5 K were reported. Polar Kerr effect measurements evidence saturation fields for perpendicular magnetization up to 15 T at low temperature, proof for a strong NiO–Co exchange interaction. The formation of a domain wall in the Co layer is a possible mechanism for such large saturation fields. © 2000 American Institute of Physics. [ABSTRACT FROM AUTHOR]

It is established that the Hall effect in Fe/SiO[sub 2] nanocomposite films in the activational tunneling conduction range is anomalous, i.e., the Hall resistivity ρ[sub h] is proportional to the magnetization and is due to the spin-orbit interaction. The parametric coupling of the Hall and longitudinal (ρ[sub xx]) resistances ρ[sub h][sup ∝]ρ[sup m, sub xx] (with temperature as the parameter) is characterized by a much lower value of the exponent m than in a uniform ferromagnetic metal. This circumstance is attributed to the characteristic features of the Hall effect mechanism in the hopping regime — in our case, the interference of the amplitudes of tunneling transitions in a set of three granules. [ABSTRACT FROM AUTHOR]

Piroplasmids are vector-borne hemoprotozoan parasites belonging to the phylum Apicomplexa that are of veterinary and medical importance. Wild carnivores are hosts for diverse piroplasmids, some of which are highly pathogenic for domestic dogs and cats. A large-scale survey including samples from 244 individuals belonging to eleven different species that were opportunistically obtained between 1993 and 2015 in four Autonomous Regions in Spain were tested for piroplasmid DNA with two different nested-PCR assays targeting the 18S rRNA gene. Sixty of 85 Eurasian badgers (Meles meles), 11 of 42 red foxes (Vulpes vulpes), and 1 of 10 stone martens (Martes foina) resulted positive. In contrast, 46 wolves (Canis lupus), 26 genets (Genetta genetta), 22 pine martens (Martes martes), and other less-represented species were negative. Sequencing revealed that all foxes and one badger were parasitized by Babesia vulpes, and the remaining badgers and the stone marten by Babesia sp. badger type A (BBTA). The prevalence of BBTA in Catalonian badgers was significantly lower in Alpine than in Continental and Mediterranean climates. This study confirms that badgers and ref foxes constitute the natural hosts of BBTA and B. vulpes, respectively, with occasional spillovers to other species., Competing Interests: Declarations. Ethical approval: The study was approved by the authorities in bioethics of Universidad de Zaragoza with reference PD12/23NE. Consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

In recent years, air sac parasitic helminths have been reported to cause severe disease in birds. In addition, various species appear to be expanding and infecting new avian hosts in various regions worldwide. In this context, an air sac nematode was initially detected in 2014 infecting the Eurasian scops owl, hospitalized in the local wildlife hospital in Mallorca (Balearic Islands, Spain). Years later, the parasite was detected in 2 other owl species. Air sac nematodes had never been reported in the Mallorcan Strigiformes before. A comprehensive molecular and morphological characterization analysis, including scanning electron microscopy, was required for species confirmation. The species was identified as Lissonema sicki , a parasite infrequently reported in South American owls. Since its first introduction to Mallorca, it has dramatically increased in prevalence in hospitalized birds, being highly prevalent in the Eurasian scops owl (41%), in the long-eared owl (11%) and in the barn owl (4%). The introduction pathway of this parasite to Europe remains unknown. This discovery underscores the expanding range and impact of L. sicki , emphasizing the importance of ongoing surveillance and research to comprehend and manage the implications of its emergence in new territories.

Neural angiostrongyliasis is an emerging zoonosis caused by the rat lungworm, Angiostrongylus cantonensis. In humans, infection with this nematode often results in eosinophilic meningitis and other severe disorders of the central nervous system. Europe was deemed a nonendemic region until 2018, when A. cantonensis worms were detected on the Mediterranean island of Mallorca, Spain, a tourism hotspot. Since that time, a sentinel surveillance system and a molecular approach have been used to follow the invasion path of the rat lungworm on the island. A. cantonensis worms have been found in animals from 8 locations on the island over 3 consecutive years. Our preliminary results show a recognizable pattern of clinical signs in infected hedgehogs and a single mitochondrial haplotype circulating in Mallorca. We present strong evidence confirming that the rat lungworm has successfully established and colonized an island in Europe and discuss observations and possible strategies for its early detection across continental Europe.

The impact of carnivore parvovirus infection on wild populations is not yet understood; disease signs are mainly developed in pups and assessing the health of litters in wild carnivores has big limitations. This study aims to shed light on the virus dynamics among wild carnivores thanks to the analysis of 213 samples collected between 1994 and 2013 in wild ecosystems from Spain. We determined the presence of carnivore parvovirus DNA by real-time PCR and sequenced the vp2 gen from 22 positive samples to characterize the strains and to perform phylogenetic analysis. The presence of carnivore parvovirus DNA was confirmed in 18% of the samples, with a higher prevalence detected in wolves (Canis lupus signatus, 70%). Fourteen sequences belonging to nine wolves, three Eurasian badgers (Meles meles), a common genet (Genetta genetta) and a European wildcat (Felis silvestris) were classified as canine parvovirus 2c (CPV-2c); five sequences from three wolves, a red fox (Vulpes vulpes) and a stone marten (Martes foina) as CPV-2b; and three sequences from a badger, a genet and a stone marten as feline parvovirus (FPV). This was the first report of a wildcat infected with a canine strain. Sequences described in this study were identical or very close related to others previously found in domestic carnivores from distant countries, suggesting that cross-species transmission takes place and that the parvovirus epidemiology in Spain, as elsewhere, could be influenced by global factors., (© 2019 Blackwell Verlag GmbH.)

In October 2018, two Atelerix algirus hedgehogs were admitted to the Wildlife Rehabilitation Hospital in Mallorca (Balearic Islands, Spain) with signs of acute neurological disease. Necropsy detected immature, fully developed nematodes in the subarachnoid space of both hedgehogs, including a gravid female worm. DNA-based molecular tools confirmed the nematode as Angiostrongylus cantonensis , an important aetiological agent of eosinophilic meningitis in humans. So far this zoonotic parasite in has not been reported in western European wildlife.

The acanthocephalan parasite Plagiorhynchus cylindraceus has a global distribution and utilizes isopods and birds as intermediate and definitive hosts, respectively. Occasionally, mammals of various orders can act as paratenic hosts. In hedgehogs, severe cases have been reported in juvenile specimens due to secondary infections, as a consequence of complete penetrations of the intestinal wall by cystacanths. In a 66-month study period, we found seven larvae of this parasite encysted in both, the peritoneal cavity and intestine of the Algerian hedgehog, Atelerix algirus in Majorca. Morphology alone was insufficient to identify the species, due to the lack of previous reports and taxonomy-informative characters. In the present report, we combined the use of morphology and the DNA-barcoding approach to confirm to identify cystacanths as P. cylindraceus. This is the first report of this parasite in this hedgehog species. The epidemiological implications will be discussed, including the risk of zoonosis and the importance of using modern approaches to identify immature acanthocephalan larvae in wildlife hosts.

Two-hundred and thirty-one wild carnivores belonging to 10 species of in Spain were analyzed for the presence of DNA of hemotropic mycoplasmas (hemoplasmas) by means of a universal real-time PCR targeting a 16S rRNA gene fragment. Positive reactions were found for wolf (Canis lupus: 6/37), fox (Vulpes vulpes: 1/41), Eurasian badger (Meles meles: 49/85), pine marten (Martes martes: 11/23), stone marten (Martes foina: 6/9), least weasel (Mustela nivalis: 4/4), European wildcat (Felis s. silvestris: 1/2) and common genet (Genetta genetta: 7/27). Sixty-four readable sequences were obtained, resulting in 14 nucleotide sequence types (ntST). The highest diversity was detected in badger (6 ntST) and pine marten (5 ntST). The sequencing of a fragment of the RNase P gene showed that all positive reactions in wolves corresponded to Mycoplasma haemocanis. Three ntST showed an identity between 98-100% with Candidatus M. haemominutum, C. M. turicensis and C. M. haematoparvum, respectively. Four ntST were closely related to C. M. haemomeles and/or diverse genotypes reported from raccoons (Procyon lotor) in the USA. One ntST from a badger showed only 88% similarity to the closest published sequence and was phylogenetically unrelated to any other hemoplasma sequence reported. Three ntST were 99-100% similar to two different sequences reported in Spanish bats. This study confirms the widespread nature and the high genetic diversity of hemoplasma infection in carnivores. Wild carnivores might be natural hosts of some hemoplasmas infecting dogs and cats., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

In the Drosophila optic lobes, 800 retinotopically organized columns in the medulla act as functional units for processing visual information. The medulla contains over 80 types of neuron, which belong to two classes: uni-columnar neurons have a stoichiometry of one per column, while multi-columnar neurons contact multiple columns. Here we show that combinatorial inputs from temporal and spatial axes generate this neuronal diversity: all neuroblasts switch fates over time to produce different neurons; the neuroepithelium that generates neuroblasts is also subdivided into six compartments by the expression of specific factors. Uni-columnar neurons are produced in all spatial compartments independently of spatial input; they innervate the neuropil where they are generated. Multi-columnar neurons are generated in smaller numbers in restricted compartments and require spatial input; the majority of their cell bodies subsequently move to cover the entire medulla. The selective integration of spatial inputs by a fixed temporal neuroblast cascade thus acts as a powerful mechanism for generating neural diversity, regulating stoichiometry and the formation of retinotopy.

The retinoblastoma (RB) tumor suppressor protein is a transcriptional cofactor with essential roles in cell cycle and development. Physical and functional targets of RB and its paralogs p107/p130 have been studied largely in cultured cells, but the full biological context of this family of proteins' activities will likely be revealed only in whole organismal studies. To identify direct targets of the major Drosophila RB counterpart in a developmental context, we carried out ChIP-Seq analysis of Rbf1 in the embryo. The association of the protein with promoters is developmentally controlled; early promoter access is globally inhibited, whereas later in development Rbf1 is found to associate with promoter-proximal regions of approximately 2000 genes. In addition to conserved cell-cycle-related genes, a wholly unexpected finding was that Rbf1 targets many components of the insulin, Hippo, JAK/STAT, Notch, and other conserved signaling pathways. Rbf1 may thus directly affect output of these essential growth-control and differentiation pathways by regulation of expression of receptors, kinases and downstream effectors. Rbf1 was also found to target multiple levels of its own regulatory hierarchy. Bioinformatic analysis indicates that different classes of genes exhibit distinct constellations of motifs associated with the Rbf1-bound regions, suggesting that the context of Rbf1 recruitment may vary within the Rbf1 regulon. Many of these targeted genes are bound by Rbf1 homologs in human cells, indicating that a conserved role of RB proteins may be to adjust the set point of interlinked signaling networks essential for growth and development.

Mitochondrial structure and function are highly dynamic, but the potential roles for cell signaling pathways in influencing these properties are not fully understood. Reduced mitochondrial function has been shown to cause cell cycle arrest, and a direct role of signaling pathways in controlling mitochondrial function during development and disease is an active area of investigation. Here, we show that the conserved Yorkie/YAP signaling pathway implicated in the control of organ size also functions in the regulation of mitochondria in Drosophila as well as human cells. In Drosophila, activation of Yorkie causes direct transcriptional up-regulation of genes that regulate mitochondrial fusion, such as opa1-like (opa1) and mitochondria assembly regulatory factor (Marf), and results in fused mitochondria with dramatic reduction in reactive oxygen species (ROS) levels. When mitochondrial fusion is genetically attenuated, the Yorkie-induced cell proliferation and tissue overgrowth are significantly suppressed. The function of Yorkie is conserved across evolution, as activation of YAP2 in human cell lines causes increased mitochondrial fusion. Thus, mitochondrial fusion is an essential and direct target of the Yorkie/YAP pathway in the regulation of organ size control during development and could play a similar role in the genesis of cancer.

Background: In holometabolous insects such as Drosophila melanogaster, neuroblasts produce an initial population of diverse neurons during embryogenesis and a much larger set of adult-specific neurons during larval life. In the ventral CNS, many of these secondary neuronal lineages differ significantly from one body segment to another, suggesting a role for anteroposterior patterning genes., Results: Here we systematically characterize the expression pattern and function of the Hox gene Ultrabithorax (Ubx) in all 25 postembryonic lineages. We find that Ubx is expressed in a segment-, lineage-, and hemilineage-specific manner in the thoracic and anterior abdominal segments. When Ubx is removed from neuroblasts via mitotic recombination, neurons in these segments exhibit the morphologies and survival patterns of their anterior thoracic counterparts. Conversely, when Ubx is ectopically expressed in anterior thoracic segments, neurons exhibit complementary posterior transformation phenotypes., Conclusion: Our findings demonstrate that Ubx plays a critical role in conferring segment-appropriate morphology and survival on individual neurons in the adult-specific ventral CNS. Moreover, while always conferring spatial identity in some sense, Ubx has been co-opted during evolution for distinct and even opposite functions in different neuronal hemilineages.

We evaluated how variations in sequencing depth and other parameters influence interpretation of chromatin immunoprecipitation-sequencing (ChIP-seq) experiments. Using Drosophila melanogaster S2 cells, we generated ChIP-seq data sets for a site-specific transcription factor (Suppressor of Hairy-wing) and a histone modification (H3K36me3). We detected a chromatin-state bias: open chromatin regions yielded higher coverage, which led to false positives if not corrected. This bias had a greater effect on detection specificity than any base-composition bias. Paired-end sequencing revealed that single-end data underestimated ChIP-library complexity at high coverage. Removal of reads originating at the same base reduced false-positives but had little effect on detection sensitivity. Even at mappable-genome coverage depth of ∼1 read per base pair, ∼1% of the narrow peaks detected on a tiling array were missed by ChIP-seq. Evaluation of widely used ChIP-seq analysis tools suggests that adjustments or algorithm improvements are required to handle data sets with deep coverage.

Behavior is among the most dynamic animal phenotypes, modulated by a variety of internal and external stimuli. Behavioral differences are associated with large-scale changes in gene expression, but little is known about how these changes are regulated. Here we show how a transcription factor (TF), ultraspiracle (usp; the insect homolog of the Retinoid X Receptor), working in complex transcriptional networks, can regulate behavioral plasticity and associated changes in gene expression. We first show that RNAi knockdown of USP in honey bee abdominal fat bodies delayed the transition from working in the hive (primarily "nursing" brood) to foraging outside. We then demonstrate through transcriptomics experiments that USP induced many maturation-related transcriptional changes in the fat bodies by mediating transcriptional responses to juvenile hormone. These maturation-related transcriptional responses to USP occurred without changes in USP's genomic binding sites, as revealed by ChIP-chip. Instead, behaviorally related gene expression is likely determined by combinatorial interactions between USP and other TFs whose cis-regulatory motifs were enriched at USP's binding sites. Many modules of JH- and maturation-related genes were co-regulated in both the fat body and brain, predicting that usp and cofactors influence shared transcriptional networks in both of these maturation-related tissues. Our findings demonstrate how "single gene effects" on behavioral plasticity can involve complex transcriptional networks, in both brain and peripheral tissues., Competing Interests: The authors have declared that no competing interests exist.

While translational stop codon readthrough is often used by viral genomes, it has been observed for only a handful of eukaryotic genes. We previously used comparative genomics evidence to recognize protein-coding regions in 12 species of Drosophila and showed that for 149 genes, the open reading frame following the stop codon has a protein-coding conservation signature, hinting that stop codon readthrough might be common in Drosophila. We return to this observation armed with deep RNA sequence data from the modENCODE project, an improved higher-resolution comparative genomics metric for detecting protein-coding regions, comparative sequence information from additional species, and directed experimental evidence. We report an expanded set of 283 readthrough candidates, including 16 double-readthrough candidates; these were manually curated to rule out alternatives such as A-to-I editing, alternative splicing, dicistronic translation, and selenocysteine incorporation. We report experimental evidence of translation using GFP tagging and mass spectrometry for several readthrough regions. We find that the set of readthrough candidates differs from other genes in length, composition, conservation, stop codon context, and in some cases, conserved stem-loops, providing clues about readthrough regulation and potential mechanisms. Lastly, we expand our studies beyond Drosophila and find evidence of abundant readthrough in several other insect species and one crustacean, and several readthrough candidates in nematode and human, suggesting that functionally important translational stop codon readthrough is significantly more prevalent in Metazoa than previously recognized.

Low-cost, high-throughput gene synthesis and precise control of protein expression are of critical importance to synthetic biology and biotechnology. Here we describe the development of an on-chip gene synthesis technology, which integrates on a single microchip the synthesis of DNA oligonucleotides using inkjet printing, isothermal oligonucleotide amplification and parallel gene assembly. Use of a mismatch-specific endonuclease for error correction results in an error rate of ~0.19 errors per kb. We applied this approach to synthesize pools of thousands of codon-usage variants of lacZα and 74 challenging Drosophila protein antigens, which were then screened for expression in Escherichia coli. In one round of synthesis and screening, we obtained DNA sequences that were expressed at a wide range of levels, from zero to almost 60% of the total cell protein mass. This technology may facilitate systematic investigation of the molecular mechanisms of protein translation and the design, construction and evolution of macromolecular machines, metabolic networks and synthetic cells.