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We report Pelecitus helicinus Railliet & Henry, 1910 from 13 species of birds of 2 orders and 7 families, collected from the states of São Paulo and Mato Grosso, Brazil. All 13 constitute new host records for this nematode. In addition, we report the first record of Aprocta golvani Diaz-Ungria, 1963 from Brazil and Monasa nigrifrons (Bucconidae), as well as a number of other nematode records from Neotropical birds.

Cell sedimentation in 3D hydrogel cultures refers to the vertical migration of cells towards the bottom of the space. Understanding this poorly examined phenomenon may allow us to design better protocols to prevent it, as well as provide insights into the mechanobiology of cancer development. We conducted a multiscale experimental and mathematical examination of 3D cancer growth in triple negative breast cancer cells. Migration was examined in the presence and absence of Paclitaxel, in high and low adhesion environments and in the presence of fibroblasts. The observed behaviour was modeled by hypothesizing active migration due to self-generated chemotactic gradients. Our results did not reject this hypothesis, whereby migration was likely to be regulated by the MAPK and TGF-β pathways. The mathematical model enabled us to describe the experimental data in absence (normalized error<40%) and presence of Paclitaxel (normalized error<10%), suggesting inhibition of random motion and advection in the latter case. Inhibition of sedimentation in low adhesion and co-culture experiments further supported the conclusion that cells actively migrated downwards due to the presence of signals produced by cells already attached to the adhesive glass surface., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Dimitriou et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Members of the genus Scaphanocephalus mature in accipitrids, particularly osprey, Pandion haliaetus , with metacercaria causing Black Spot Syndrome in reef fishes. In most of the world, only the type species, Scaphanocephalus expansus (Creplin, 1842) has been reported. Recent molecular studies in the Western Atlantic, Mediterranean and Persian Gulf reveal multiple species of Scaphanocephalus , but have relied on 28S rDNA, mainly from metacercariae, which limits both morphological identification and resolution of closely related species. Here we combine nuclear rDNA with mitochondrial sequences from adult worms collected in osprey across North America and the Caribbean to describe species and elucidate life cycles in Scaphanocephalus . A new species described herein can be distinguished from S. expansus based on overall body shape and size. Phylogenetic analysis of the whole mitochondrial genome of Scaphanocephalus indicates a close relationship with Cryptocotyle . We conclude that at least 3 species of Scaphanocephalus are present in the Americas and 2 others are in the Old World. Specimens in the Americas have similar or identical 28S to those in the Mediterranean and Persian Gulf, but amphi-Atlantic species are unlikely in light of divergence in cytochrome c oxidase I and the lack of amphi-Atlantic avian and fish hosts. Our results provide insight into the geographic distribution and taxonomy of a little-studied trematode recently linked to an emerging pathology in ecologically important reef fishes.

A parasite-host, host-parasite and distribution-based checklist of helminths found in bats (Chiroptera) of North America north of Mexico is presented. The parasite-host checklist includes a total of 93 species (including records without a species identification) of helminth parasites reported in the literature from 30 species of bats. These include 54 trematodes, 11 cestodes, and 28 nematodes. Each helminth species is listed under its most current accepted name, with all known synonyms, distribution by state/province, and references for each geographic location. Lists of helminths reported from individual species of bats as well as states of the United States and provinces/territories of Canada are also provided. The following new combinations are proposed: Paralecithodendrium alaskensis (Neiland, 1962) n. comb. for Prosthodendrium alaskensis Neiland, 1962; Paralecthodendrium longiforme (Bhalerao, 1926) n. comb. for Lecithodendrium longiforme Bhalerao, 1926; and Paralecithodendrium singularium (Byrd & Macy, 1942) n. comb. for Prosthodendrium singularium Byrd & Macy, 1942. The state of knowledge of helminths of bats in North America is briefly discussed.

The engineering of a new monodisperse colloid with a sea urchin-like structure with a large complex internal structure is reported, in which silica surfaces are bridged by an aromatic organic cross-linker to serve as a nanocarrier host for drugs such as doxorubicin (DOX) against breast cancer cells. While dendritic fibrous nanosilica (DFNS) was employed and we do not observe a dendritic structure, these particles are referred to as sea urchin-like nanostructured silica (SNS). Since the structure of SNS consists of many silica fibrils protruding from the core, similar to the hairs of a sea urchin. For the aromatic structured cross-linker, bis(propyliminomethyl)benzene (b(PIM)B-S or silanated terephtaldehyde) were employed, which are prepared with terephtaldehyde and 3-aminopropyltriethoxy-silane (APTES) through a simple Schiff base reaction. b(PIM)B-S bridges were introduced into SNS under open vessel reflux conditions. SPS refers to the product obtained by incorporating the cross-linker b(PIM)B-S in ultra-small colloidal SNS particles. In-situ incorporation of DOX molecules resulted in SPS-DOX. The pH-responsive SPS nanocomposites were tested as biocompatible nanocarriers for controllable doxorubicin (DOX) delivery. We conclude that SPS is a unique colloid which has promising potential for technological applications such as advanced drug delivery systems, wastewater remediation and as a catalyst for green organic reactions in water., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Inc.)

The immune response against a tumor is characterized by the interplay among components of the immune system and neoplastic cells. Here, we bioprinted a model with two distinct regions containing gastric cancer patient-derived organoids (PDOs) and tumor-infiltrated lymphocytes (TILs). The initial cellular distribution allows for the longitudinal study of TIL migratory patterns concurrently with multiplexed cytokine analysis. The chemical properties of the bioink were designed to present physical barriers that immune T-cells must breech during infiltration and migration toward a tumor with the use of an alginate, gelatin, and basal membrane mix. TIL activity, degranulation, and regulation of proteolytic activity reveal insights into the time-dependent biochemical dynamics. Regulation of the sFas and sFas-ligand present on PDOs and TILs, respectively, and the perforin and granzyme longitudinal secretion confirms TIL activation when encountering PDO formations. TIL migratory profiles were used to create a deterministic reaction-advection diffusion model. The simulation provides insights that decouple passive from active cell migration mechanisms. The mechanisms used by TILs and other adoptive cell therapeutics as they infiltrate the tumor barrier are poorly understood. This study presents a pre-screening strategy for immune cells where motility and activation across ECM environments are crucial indicators of cellular fitness.

Mathematical models of cancer growth have become increasingly more accurate both in the space and time domains. However, the limited amount of data typically available has resulted in a larger number of qualitative rather than quantitative studies. In the present study, we provide an integrated experimental-computational framework for the quantification of the morphological characteristics and the mechanistic modelling of cancer progression in 3D environments. The proposed framework allows for the calibration of multiscale, spatiotemporal models of cancer growth using state-of-the-art 3D cell culture data, and their validation based on the resulting experimental morphological patterns using spatial point-pattern analysis techniques. We applied this framework to the study of the development of Triple Negative Breast Cancer cells cultured in Matrigel scaffolds, and validated the hypothesis of chemotactic migration using a multiscale, hybrid Keller-Segel model. The results revealed transient, non-random spatial distributions of cancer cells that consist of clustered, and dispersion patterns. The proposed model was able to describe the general characteristics of the experimental observations and suggests that chemotactic migration together with random motion was found to be a plausible mechanism leading to accumulation, during the examined time period of development. The developed framework enabled us to pursue two goals; first, the quantitative description of the morphology of cancer growth in 3D cultures using point-pattern analysis, and second, the relation of tumour morphology with underlying biophysical mechanisms that govern cancer growth and migration.

Malignant tumor tissues exhibit inter- and intratumoral heterogeneities, aberrant development, dynamic stromal composition, diverse tissue phenotypes, and cell populations growing within localized mechanical stresses in hypoxic conditions. Experimental tumor models employing engineered systems that isolate and study these complex variables using in vitro techniques are under development as complementary methods to preclinical in vivo models. Here, advances in extrusion bioprinting as an enabling technology to recreate the three-dimensional tumor milieu and its complex heterogeneous characteristics are reviewed. Extrusion bioprinting allows for the deposition of multiple materials, or selected cell types and concentrations, into models based upon physiological features of the tumor. This affords the creation of complex samples with representative extracellular or stromal compositions that replicate the biology of patient tissue. Biomaterial engineering of printable materials that replicate specific features of the tumor microenvironment offer experimental reproducibility, throughput, and physiological relevance compared to animal models. In this review, we describe the potential of extrusion-based bioprinting to recreate the tumor microenvironment within in vitro models.

The invasion of cancer cells into the surrounding tissues is one of the hallmarks of cancer. However, a precise quantitative understanding of the spatiotemporal patterns of cancer cell migration and invasion still remains elusive. A promising approach to investigate these patterns are 3D cell cultures, which provide more realistic models of cancer growth compared to conventional 2D monolayers. Quantifying the spatial distribution of cells in these 3D cultures yields great promise for understanding the spatiotemporal progression of cancer. In the present study, we present an image processing and segmentation pipeline for the detection of 3D GFP-fluorescent triple-negative breast cancer cell nuclei, and we perform quantitative analysis of the formed spatial patterns and their temporal evolution. The performance of the proposed pipeline was evaluated using experimental 3D cell culture data, and was found to be comparable to manual segmentation, outperforming four alternative automated methods. The spatiotemporal statistical analysis of the detected distributions of nuclei revealed transient, non-random spatial distributions that consisted of clustered patterns across a wide range of neighbourhood distances, as well as dispersion for larger distances. Overall, the implementation of the proposed framework revealed the spatial organization of cellular nuclei with improved accuracy, providing insights into the 3 dimensional inter-cellular organization and its progression through time., (© 2022. The Author(s) under exclusive licence to Biomedical Engineering Society.)

Long-term data allow ecologists to assess trajectories of population abundance. Without this context, it is impossible to know whether a taxon is thriving or declining to extinction. For parasites of wildlife, there are few long-term data-a gap that creates an impediment to managing parasite biodiversity and infectious threats in a changing world. We produced a century-scale time series of metazoan parasite abundance and used it to test whether parasitism is changing in Puget Sound, United States, and, if so, why. We performed parasitological dissection of fluid-preserved specimens held in natural history collections for eight fish species collected between 1880 and 2019. We found that parasite taxa using three or more obligately required host species-a group that comprised 52% of the parasite taxa we detected-declined in abundance at a rate of 10.9% per decade, whereas no change in abundance was detected for parasites using one or two obligately required host species. We tested several potential mechanisms for the decline in 3+-host parasites and found that parasite abundance was negatively correlated with sea surface temperature, diminishing at a rate of 38% for every 1 °C increase. Although the temperature effect was strong, it did not explain all variability in parasite burden, suggesting that other factors may also have contributed to the long-term declines we observed. These data document one century of climate-associated parasite decline in Puget Sound-a massive loss of biodiversity, undetected until now.

Alaria , Didelphodiplostomum and Pharyngostomoides are among genera of diplostomid digeneans known to parasitize mammalian definitive hosts. Despite numerous recent molecular phylogenetic studies of diplostomids, limited DNA sequence data is available from diplostomids parasitic in mammals. Herein, we provide the first 28S rDNA and cox1 mtDNA sequences from morphologically identified, adult specimens of Didelphodiplostomum and Pharyngostomoides . Newly generated 28S sequences were used to infer the phylogenetic interrelationships of these two genera among other major lineages of diplostomoideans. The phylogeny based on 28S and a review of morphology clearly suggests that Pharyngostomoides should be considered a junior synonym of Alaria , while Didelphodiplostomum should be considered a junior synonym of Tylodelphys . Pharyngostomoides procyonis (type species), Pharyngostomoides adenocephala and Pharyngostomoides dasyuri were transferred into Alaria as Alaria procyonis comb. nov. , Alaria adenocephala comb. nov. and Alaria dasyuri comb. nov. ; Didelphodiplostomum variabile (type species) and Didelphodiplostomum nunezae were transferred into Tylodelphys as Tylodelphys variabilis comb. nov. and Tylodelphys nunezae comb. nov. In addition, Alaria ovalis comb. nov. (formerly included in Pharyngostomoides ) was restored and transferred into Alaria based on a morphological study of well-fixed, adult specimens and the comparison of cox1 DNA sequences among Alaria spp. The diplostomid genus Parallelorchis was restored based on review of morphology., (© The Author(s) 2022. Published by Oxford University Press on behalf of The Linnean Society of London. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Oxyurid nematodes (Syphacia spp.) from bank (Myodes glareolus) and field/common (Microtus spp.) voles, from disparate geographical sites in the British Isles, were examined morphologically and genetically. The genetic signatures of 118 new isolates are provided, based primarily on the rDNA internal transcribed spacers (ITS1-5.8S-ITS2) region and for representative isolates also on the small subunit 18S rDNA region and cytochrome c oxidase subunit 1 (cox-1) gene locus. Genetic data on worms recovered from Microtus spp. from the European mainland and from other rodent genera from the Palaearctic, North America and West Africa are also included. We test historical hypotheses indicating that S. nigeriana is a generalist species, infecting a range of different rodent genera. Our results establish that S. nigeriana is a parasite of both bank and field voles in the British Isles. An identical genotype was also recorded from Hubert's multimammate mouse (Mastomys huberti) from Senegal, but Mastomys spp. from West Africa were additionally parasitized by a related, although genetically distinct Syphacia species. We found no evidence for S. petrusewiczi in voles from the British Isles but isolates from Russia and North America were genetically distinct and formed their own separate deep branch in maximum likelihood molecular phylogenetic trees.

Reinforced extracellular matrix (ECM)-based hydrogels recapitulate several mechanical and biochemical features found in the tumor microenvironment (TME) in vivo. While these gels retain several critical structural and bioactive molecules that promote cell-matrix interactivity, their mechanical properties tend toward the viscous regime limiting their ability to retain ordered structural characteristics when considered as architectured scaffolds. To overcome this limitation characteristic of pure ECM hydrogels, we present a composite material containing alginate, a seaweed-derived polysaccharide, and gelatin, denatured collagen, as rheological modifiers which impart mechanical integrity to the biologically active decellularized ECM (dECM). After an optimization process, the reinforced gel proposed is mechanically stable and bioprintable and has a stiffness within the expected physiological values. Our hydrogel's elastic modulus has no significant difference when compared to tumors induced in preclinical xenograft head and neck squamous cell carcinoma (HNSCC) mouse models. The bioprinted cell-laden model is highly reproducible and allows proliferation and reorganization of HNSCC cells while maintaining cell viability above 90% for periods of nearly 3 weeks. Cells encapsulated in our bioink produce spheroids of at least 3000 μm 2 of cross-sectional area by day 15 of culture and are positive for cytokeratin in immunofluorescence quantification, a common marker of HNSCC model validation in 2D and 3D models. We use this in vitro model system to evaluate the standard-of-care small molecule therapeutics used to treat HNSCC clinically and report a 4-fold increase in the IC 50 of cisplatin and an 80-fold increase for 5-fluorouracil compared to monolayer cultures. Our work suggests that fabricating in vitro models using reinforced dECM provides a physiologically relevant system to evaluate malignant neoplastic phenomena in vitro due to the physical and biological features replicated from the source tissue microenvironment.

Equids are chronically infected with parasitic strongyle nematodes. There is a rich literature on horse strongyles, but they are difficult to identify morphologically and genetic studies on strongyles infecting other equid species are few, hampering studies of host specificity. We sequenced expelled worms from two sympatric zebra species in central Kenya to expand the strongyle phylogeny and used DNA metabarcoding on faecal samples to genetically characterize zebra nemabiomes for the first time. We generated sequences for several species new to public genetic reference databases, all of which are typical strongyles in wild zebras (i.e., the three species of Cylindropharynx and Cyathostomum montgomeryi ), and identified their closest relatives. We also discovered an apparent fungus infecting a quarter of the expelled Crossocephalus viviparus worms, a hyperabundant nematode species in the family Atractidae, hinting at the possibility that zebra host-parasite dynamics may involve a zebra-fungus mutualism. The two zebra species had similar nemabiomes; we found a complete overlap in the list of nematode species they carry and very similar prevalence (i.e., proportion of hosts infected) for the different nematode species. Our study suggests limited host-specificity in zebra strongyles and high potential for transmission between the plains zebra and the endangered Grevy's zebra., Competing Interests: None., (© 2021 Published by Elsevier Ltd on behalf of Australian Society for Parasitology.)

Invasive species impact native wildlife in several ways, as they compete for resources and may transmit their specific pathogens. However, the potential consequences of co-introduced parasites are not fully understood. While the house sparrow (Passer domesticus) was introduced in Chile about a century ago, no data are available regarding its parasites. Thus, the aim of the present study was to determine the parasitic fauna of this avian invader and to determine whether there are co-introduced/co-invasive parasites shared with native birds. One hundred and eight birds were collected from three different localities in the Ñuble region of Chile, and a complete parasitic necropsy was performed in the laboratory. Twenty-three (21.3%) were parasitized by six arthropod species and four (3.7%) were parasitized by two helminth species. Four out of eight taxa are reported for the first time in Chile; among them, three arthropod parasites and the tapeworm, Anonchotaenia globate, are considered as co-introduced parasites. Only A. globata is a potential co-invasive parasite given its low degree specificity in terms of its definitive hosts. Future research should examine whether additional co-introduced/co-invasive parasites have been brought by the house sparrow, and what their potential consequences might be on the health of native birds in Chile.

Purpose: Nothing is known about the coccidian parasites of vagrant shrews, Sorex vagrans Baird, 1868. Here we report, for the first time, the occurrence of Eimeria longirostris Hertel and Duszynski, 1987 from faecal contents of S. vagrans from Montana, USA., Methods: Faecal samples, collected in July and August 2020 from six pitfall-trapped vagrant shrews as well as faeces from two masked shrews, Sorex cinereus Kerr, 1792, and one American pygmy shrew, Sorex hoyi Baird, 1857 from Missoula County, Montana, USA, were examined for coccidian parasites. Samples were placed in individual vials containing aqueous potassium dichromate. They were examined for coccidia after flotation in Sheather's sugar solution, measured, and photographed., Results: Three (50%) S. vagrans and one (50%) S. cinereus were found to be passing oocysts of Eimeria longirostris Hertel and Duszynski, 1987; the single S. hoyi was negative. Oocysts from S. vagrans were subspheroidal and measured (average L × W) 16.1 × 14.4 µm with an L/W ratio of 1.1. One (typically) to two polar granules was present but a micropyle and oocyst residuum were absent. Sporocysts were ovoidal and measured 9.6 × 6.2 µm with an L/W ratio of 1.6. A Stieda body was present but subStieda and paraStieda bodies were absent. The sporocyst residuum was composed of various sized granules typically scattered between and across the sporozoites but sometimes formed a loose aggregate or compact mass., Conclusion: We document a new host and new geographic record for E. longirostris from S. vagrans and report the coccidian from S. cinereus for the third time but the first report from specimens from Montana. This coccidian has now been reported from at least 12 species of shrews within the genus Sorex in 14 US states and two provinces in Canada.

We designed three types of hollow-shaped porous silica materials via a three-step biotemplate-directed method: porous hollow silica nanorods, hollow dendritic fibrous nanostructured silica (DFNS), and ultraporous sponge-like DFNS. The first step was making a biotemplate, for which we used cellulose nanocrystals (CNCs), consisting of rod-shaped nanoparticles synthesized by conventional acid hydrolysis of cellulose fibers. In a second step, core-shell samples were prepared using CNC particles as hard template by two procedures. In the first one, core-shell CNC-silica nanoparticles were synthesized by a polycondensation reaction, which exclusively took place at the surface of the CNCs. In the second procedure, a typical synthesis of DFNS was conducted in a bicontinuous microemulsion with the assistance of additives. DFNS was assembled on the surface of the CNCs, giving rise to core-shell CNC-DFNS structures. Finally, all of the silica-coated CNC composites were calcined, during which the CNC was removed from the core and hollow structures were formed. These materials are very lightweight and highly porous. All three structures were tested as nanocarriers for drug delivery and absorbents for dye removal applications. Dye removal results showed that they can adsorb methylene blue efficiently, with ultraporous sponge-like DFNS showing the highest adsorption capacity, followed by hollow DFNS and hollow silica nanorods. Furthermore, breast cancer cells show a lower cell viability when exposed to doxorubicin-loaded hollow silica nanorods compared with control or doxorubicin cultures, suggesting that the loaded nanorod has a greater anticancer effect than free doxorubicin.

Hydrogels consisting of controlled fractions of alginate, gelatin, and Matrigel enable the development of patient-derived bioprinted tissue models that support cancer spheroid growth and expansion. These engineered models can be dissociated to be then reintroduced to new hydrogel solutions and subsequently reprinted to generate multigenerational models. The process of harvesting cells from 3D bioprinted models is possible by chelating the ions that crosslink alginate, causing the gel to weaken. Inclusion of the gelatin and Matrigel fractions to the hydrogel increases the bioactivity by providing cell-matrix binding sites and promoting cross-talk between cancer cells and their microenvironment. Here we show that immortalized triple-negative breast cancer cells (MDA-MB-231) and patient-derived gastric adenocarcinoma cells can be reprinted for at least three 21 d culture cycles following bioprinting in the alginate/gelatin/Matrigel hydrogels. Our drug testing results suggest that our 3D bioprinted model can also be used to recapitulate in vivo patient drug response. Furthermore, our results show that iterative bioprinting techniques coupled with alginate biomaterials can be used to maintain and expand patient-derived cancer spheroid cultures for extended periods without compromising cell viability, altering division rates, or disrupting cancer spheroid formation., (© 2021 IOP Publishing Ltd.)

Hydrogels have been used for a variety of biomedical applications; in tissue engineering, they are commonly used as scaffolds to cultivate cells in a three-dimensional (3D) environment allowing the formation of organoids or cellular spheroids. Egg white-alginate (EWA) is a novel hydrogel which combines the advantages of both egg white and alginate; the egg white material provides extracellular matrix (ECM)-like proteins that can mimic the ECM microenvironment, while alginate can be tuned mechanically through its ionic crosslinking property to modify the scaffold's porosity, strength, and stiffness. In this study, a frozen calcium chloride (CaCl 2 ) disk technique to homogenously crosslink alginate and egg white hydrogel is presented for 2.5D culture of human salivary cells. Different EWA formulations were prepared and biologically evaluated as a spheroid-like structure platform. Although all five EWA hydrogels showed biocompatibility, the EWA with 1.5% alginate presented the highest cell viability, while EWA with 3% alginate promoted the formation of larger size salivary spheroid-like structures. Our EWA hydrogel has the potential to be an alternative 3D culture scaffold that can be used for studies on drug-screening, cell migration, or as an in vitro disease model. In addition, EWA can be used as a potential source for cell transplantation (i.e., using this platform as an ex vivo environment for cell expansion). The low cost of producing EWA is an added advantage.

Parasitological studies of the Franklin's gull, Leucophaeus pipixcan, are scarce, and knowledge about its endoparasites is quite limited. In order to describe its parasitic community, a total of 60 Franklin's gulls were captured in the coastal area in central Chile, using modified Bal-chatri traps. Ectoparasites were collected from all 60 live individuals through inspection of their plumage, while 30 were examined for endoparasites by standard parasitological necropsy. The prevalence of ectoparasites was 78.3%, including the feather mite Zachvatkinia larica (43.3%) and four species of lice: Actornithophilus piceus lari (15.0%), Austromenopon transversum (6.7%), Quadraceps punctatus (10.0%) and Saemundssonia lari (46.7%). Some 25 of 30 (83.3%) of birds necropsied were parasitized with the following helminths: Aporchis sp. (6.7%), Tetrabothrius cylindraceus (56.7%), Cyclophyllidea gen. sp. (3.3%), Profilicollis altmani (56.7%), Eucoleus contortus (10.0%), Cosmocephalus obvelatus (13.3%), Paracuaria adunca (10.0%), Stegophorus sp. (3.3%) and Tetrameres skrjabini (3.3%). To our knowledge, with the exception of P. altmani, these helminths are reported for first time in the Franklin's gull.

The Neotropic cormorant Nannopterum (Phalacrocorax) brasilianus (Suliformes: Phalacrocoracidae) is widely distributed in Central and South America. In Chile, information about parasites for this species is limited to helminths and nematodes, and little is known about other parasite groups. This study documents the parasitic fauna present in 80 Neotropic cormorants' carcasses collected from 2001 to 2008 in Antofagasta, Biobío, and Ñuble regions. Birds were externally inspected for ectoparasites and necropsies were performed to examine digestive and respiratory organs in search of endoparasites. Ectoparasites collected were cleared and mounted for identification under a microscope. Fecal samples were also evaluated to determine the presence of protozoan parasites employing a flotation technique. A total of 44 (42.5%) of birds were infested with at least one ectoparasite species, while 77 (96.25%) were carrying endoparasites. No protozoan forms were found after examination. Most prevalent endoparasite species found were Contracaecum rudolphii s. l. (72/80, 90%), followed by Pectinopygus gyroceras (33/80, 41.25%), and Profilicollis altmani (26/80, 32.5%). This is the first report of P. altmani, Baruscapillaria carbonis, Avioserpens sp., Cyathostoma (Cyathostoma) phenisci, and Eidmaniella pelucida in the Neotropic cormorant. These findings also expand the distributional range of Andracantha phalacrocoracis, Paradilepis caballeroi, Hysteromorpha triloba, and P. gyroceras to Chile.

In this study, a carboxyl-modified cellulosic hydrogel was developed as the base material for wound dressings. ε-poly-l-lysine, a natural polyamide, was then covalently linked to the hydrogel through a bioconjugation reaction, which was confirmed by X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FTIR). The antibacterial efficacy of the hydrogel was tested against two model bacteria, Staphylococcus aureus and Pseudomonas aeruginosa , two of the most commonly found bacteria in wound infections. Bacterial viability and biofilm formation after exposure of bacteria to the hydrogels were used as efficacy indicators. Live/Dead assay was used to measure the number of compromised bacteria using a confocal laser scanning microscope. The results show that the antibacterial hydrogel was able to kill approximately 99% of the exposed bacteria after 3 h of exposure. In addition, NIH/3T3 fibroblasts were used to study the biocompatibility of the developed hydrogels. Water-soluble tetrazolium salt (WST)-1 assay was used to measure the metabolic activity of the cells and Live/Dead assay was used to measure the viability of the cells after 24, 48, and 72 h. The developed antibacterial hydrogels are light weight, have a high water-uptake capacity, and show high biocompatibility with the model mammalian cells, which make them a promising candidate to be used for wound dressing applications.

Cell-laden scaffolds of architecture and mechanics that mimic those of the host tissues are important for a wide range of biomedical applications but remain challenging to bioprint. To address these challenges, we report a new method called triggered micropore-forming bioprinting. The approach can yield cell-laden scaffolds of defined architecture and interconnected pores over a range of sizes, encompassing that of many cell types. The viscoelasticity of the bioprinted scaffold can match that of biological tissues and be tuned independently of porosity and stiffness. The bioprinted scaffold also exhibits superior mechanical robustness despite high porosity. The bioprinting method and the resulting scaffolds support cell spreading, migration, and proliferation. The potential of the 3D bioprinting system is demonstrated for vocal fold tissue engineering and as an in vitro cancer model. Other possible applications are foreseen for tissue repair, regenerative medicine, organ-on-chip, drug screening, organ transplantation, and disease modeling., Competing Interests: Conflicts of interest There are no conflicts to declare.

Comparative studies of parasites in sympatric bird species have been generally scarce. Parasitic infection/transmission can be spread in a number of ways that suggests possible direct and indirect, horizontal transmission between avian hosts. In order to determine whether two sympatric icterids from Central and Southern Chile share their parasite fauna (ecto- and endoparasites), we examined parasites of 27 Shiny Cowbirds, Molothrus bonariensis, and 28 Austral Blackbirds, Curaeus curaeus, including individuals captured in the wild and carcasses. We found that Shiny Cowbirds were infected with the chewing lice Brueelia bonariensis, Philopterus sp. 1, the feather mites Amerodectes molothrus, Proctophyllodes spp. (species 1 and 2), and the helminths Mediorhynchus papillosus, Plagiorhynchus sp., Dispharynx nasuta and Tetrameres paucispina, while Austral Blackbirds had the chewing lice Myrsidea sp., Philopterus sp. 2, the feather mites Proctophyllodes sp. 3, Amerodectes sp., and three helminths: Anonchotaenia sp., Capillaria sp. and M. papillosus. The flea Dasypsyllus (Neornipsyllus) cteniopus was found only on the Austral Blackbird. The only parasite species shared by both icterids was the acanthocephalan M. papillosus, possibly due to their feeding on the same intermediate insect hosts. With the exception of B. bonariensis and Philopterus sp. 1 found on the Shiny Cowbird, all species reported in this study represent new parasite-host associations and new records of parasite diversity in Chile.

Tunable bioprinting materials are capable of creating a broad spectrum of physiological mimicking 3D models enabling in vitro studies that more accurately resemble in vivo conditions. Tailoring the material properties of the bioink such that it achieves both bioprintability and biomimicry remains a key challenge. Here we report the development of engineered composite hydrogels consisting of gelatin and alginate components. The composite gels are demonstrated as a cell-laden bioink to build 3D bioprinted in vitro breast tumor models. The initial mechanical characteristics of each composite hydrogel are correlated to cell proliferation rates and cell spheroid morphology spanning month long culture conditions. MDA-MB-231 breast cancer cells show gel formulation-dependency on the rates and frequency of self-assembly into multicellular tumor spheroids (MCTS). Hydrogel compositions comprised of decreasing alginate concentrations, and increasing gelatin concentrations, result in gels that are mechanically soft and contain a greater number of cell-adhesion moieties driving the development of large MCTS; conversely gels containing increasing alginate, and decreasing gelatin concentrations are mechanically stiffer, with fewer cell-adhesion moieties present in the composite gels yielding smaller and less viable MCTS. These composite hydrogels can be used in the biofabrication of tunable in vitro systems that mimic both the mechanical and biochemical properties of the native tumor stroma.

For salivary gland (SG) tissue engineering, we cultured acinar NS-SV-AC cell line or primary SG fibroblasts for 14 days in avian egg yolk plasma (EYP). Media or egg white (EW) supplemented the cultures as they grew in 3D-Cryo histology well inserts. In the second half of this manuscript, we measured EYP's freeze-thaw gelation and freeze-thaw induced gelled EYP ( G EYP), and designed and tested further G EYP tissue engineering applications. With a 3D-Cryo well insert, we tested G EYP as a structural support for 3D cell culture or as a bio-ink for 3D-Bioprinting fluorescent cells. In non-printed EYP + EW or G EYP + EW cultures, sagittal sections of the cultures showed cells remaining above the well's base. Ki-67 expression was lacking for fibroblasts, contrasting NS-SV-AC's constant expression. Rheological viscoelastic measurements of G EYP at 37 °C on seven different freezing periods showed constant increase from 0 in mean storage and loss moduli, to 320 Pa and 120 Pa, respectively, after 30 days. We successfully 3D-printed G EYP with controlled geometries. We manually extruded G EYP bio-ink with fluorescence cells into a 3D-Cryo well insert and showed cell positioning. The 3D-Cryo well inserts reveal information on cells in EYP and we demonstrated G EYP cell culture and 3D-printing applications.

A total of thirty Austral thrushes Turdus falcklandii Quoy & Gaimard, 1824 (Turdidae) carcasses were brought to the Departamento de Ciencia Animal, Facultad de Ciencias Veterinarias, Universidad de Concepción, to be examined for ecto- and endoparasites. Ectoparasites were found on 20% (6/30) of the thrushes and belonged to species Brueelia magellanica Cichino, 1986 (Phthiraptera), Menacanthus eurysternus Burmeister, 1838 (Phthiraptera) and Tyrannidectes falcklandicus Mironov & González-Acuña, 2011 (Acari). Endoparasites were isolated from 26.6% (8/30) of the birds and identified as Lueheia inscripta Westrumb, 1821 (Acanthocephala), Plagiorhynchus cylindraceus Goeze, 1782 (Acanthocephala), Wardium sp. sensu Mayhew, 1925 (Cestoda), Dilepis undula (Cestoda) Schrank, 1788, and Zonorchis sp. (sensu Travassos, 1944) (Trematoda). To our knowledge, all endoparasites collected in this study are new records in T. falcklandii and expand their distributional range to Chile.

Information about parasites associated with diurnal raptors from Chile is scarce. Between 2006 and 2017, a total of 15 specimens of the Variable hawk, Geranoaetus polyosoma (Quoy & Gaimard, 1824) were collected, 14 of them from different localities in the Biobío region and one specimen from the Valparaíso region. An external examination of the plumage was made to collect ectoparasites, and necropsies were performed, focusing primarily on the gastrointestinal tract. Chewing lice (Phthiraptera) were found on five (33.3%) of the birds corresponding to three species: 97 specimens of Degeeriella fulva (Giebel, 1874), six specimens of Colpocephalum turbinatum Denny, 1842 and nine belonging to an unidentified species of the genus Craspedorrhynchus Kéler, 1938. Endoparasites found in three (20%) of the birds included round worms (Nematoda) of the genus Procyrnea Chabaud, 1958, and spiny-headed worms (Acanthocephala) of the genus Centrorhynchus Lühe, 1911. The species Colpocephalum turbinatum and the genera: Craspedorrhynchus sp., Procyrnea sp. and Centrorhynchus sp. are new records for the Variable hawk.

Three-dimensional (3D) printing is an emerging technology in the field of dentistry. It uses a layer-by-layer manufacturing technique to create scaffolds that can be used for dental tissue engineering applications. While several 3D printing methodologies exist, such as selective laser sintering or fused deposition modeling, this paper will review the applications of 3D printing for craniofacial tissue engineering; in particular for the periodontal complex, dental pulp, alveolar bone, and cartilage. For the periodontal complex, a 3D printed scaffold was attempted to treat a periodontal defect; for dental pulp, hydrogels were created that can support an odontoblastic cell line; for bone and cartilage, a polycaprolactone scaffold with microspheres induced the formation of multiphase fibrocartilaginous tissues. While the current research highlights the development and potential of 3D printing, more research is required to fully understand this technology and for its incorporation into the dental field.

Immunomodulation strategies are believed to improve the integration and clinical performance of synthetic bone substitutes. One potential approach is the modification of biomaterial surface chemistry to mimic bone extracellular matrix (ECM). In this sense, we hypothesized that coating synthetic dicalcium phosphate (DCP) bioceramics with bone ECM proteins would modulate the host immune reactions and improve their regenerative performance. To test this, we evaluated the in vitro proteomic surface interactions and the in vivo performance of ECM-coated bioceramic scaffolds. Our results demonstrated that coating DCP scaffolds with bone extracts, specifically those containing calcium-binding proteins, dramatically modulated their interaction with plasma proteins in vitro, especially those relating to the innate immune response. In vivo, we observed an attenuated inflammatory response against the bioceramic scaffolds and enhanced peri-scaffold new bone formation supported by the increased osteoblastogenesis and reduced osteoclastogenesis. Furthermore, the bone extract rich in calcium-binding proteins can be 3D-printed to produce customized hydrogels with improved regeneration capabilities. In summary, bone extracts containing calcium-binding proteins can enhance the integration of synthetic biomaterials and improve their ability to regenerate bone probably by modulating the host immune reaction. This finding helps understand how bone allografts regenerate bone and opens the door for new advances in tissue engineering and bone regeneration. STATEMENT OF SIGNIFICANCE: Foreign-body reaction is an important determinant of in vivo biomaterial integration, as an undesired host immune response can compromise the performance of an implanted biomaterial. For this reason, applying immunomodulation strategies to enhance biomaterial engraftment is of great interest in the field of regenerative medicine. In this article, we illustrated that coating dicalcium phosphate bioceramic scaffolds with bone-ECM extracts, especially those rich in calcium-binding proteins, is a promising approach to improve their surface proteomic interactions and modulate the immune responses towards such biomaterials in a way that improves their bone regeneration performance. Collectively, the results of this study may provide a conceivable explanation for the mechanisms involved in presenting the excellent regenerative efficacy of natural bone grafts., (Copyright © 2019 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

The cellular, biochemical, and biophysical heterogeneity of the native tumor microenvironment is not recapitulated by growing immortalized cancer cell lines using conventional two-dimensional (2D) cell culture. These challenges can be overcome by using bioprinting techniques to build heterogeneous three-dimensional (3D) tumor models whereby different types of cells are embedded. Alginate and gelatin are two of the most common biomaterials employed in bioprinting due to their biocompatibility, biomimicry, and mechanical properties. By combining the two polymers, we achieved a bioprintable composite hydrogel with similarities to the microscopic architecture of a native tumor stroma. We studied the printability of the composite hydrogel via rheology and obtained the optimal printing window. Breast cancer cells and fibroblasts were embedded in the hydrogels and printed to form a 3D model mimicking the in vivo microenvironment. The bioprinted heterogeneous model achieves a high viability for long-term cell culture (> 30 days) and promotes the self-assembly of breast cancer cells into multicellular tumor spheroids (MCTS). We observed the migration and interaction of the cancer-associated fibroblast cells (CAFs) with the MCTS in this model. By using bioprinted cell culture platforms as co-culture systems, it offers a unique tool to study the dependence of tumorigenesis on the stroma composition. This technique features a high-throughput, low cost, and high reproducibility, and it can also provide an alternative model to conventional cell monolayer cultures and animal tumor models to study cancer biology.

With the aim to identify the parasite fauna of plumbeous rail, Pardirallus sanguinolentus (Aves: Rallidae) in Chile, 26 carcasses were parasitologically necropsied. The present study revealed the presence of 14 species of parasites (inverse Simpson index = 4.64; evenness = 0.332), including ectoparasites: feather mites: Analloptes megnini , Grallobia sp., Grallolichus sp., Megniniella sp., and Metanalges sp.; the feather lice Pseudomenopon meinertzhageni, Rallicola andinus, and Fulicoffula sp.; and six species of gastrointestinal helminths: Heterakis psophiae, Porrocaecum ardeae, Tetrameres sp., Capillaria sp., Diorchis sp., and Plagiorhynchus sp. The relatively high parasite richness that was found could be attributed to the highly favorable conditions of wetlands for parasite development. All parasites found, except feather lice, are new records for plumbeous rail. A checklist of parasites for plumbeous rail is presented.

Two new species of Staphylocystoides are described from masked shrews Sorex cinereus. Staphylocystoides oligospinosus n. sp. was collected from the vicinity of Missoula, Montana, United States, and Staphylocystoides parasphenomorphus n. sp. was collected from the vicinity of Whitehorse, Yukon Territory, Canada. Morphological differentiation from known species is provided. Both species are morphologically closest to Staphylocystoides sphenomorphus and to each other. Among other characters, S. oligospinosus n. sp. can be easily differentiated from all known species of the genus by unique cirrus armature, which consists of a short zone of small spines at the base of the cirrus, a few large sparsely distributed spines of varying size in the middle part of the cirrus, and hair-like microtriches densely covering the apical portion of the cirrus. Staphylocystoides parasphenomorphus n. sp. differs from S. oligospinosus n. sp. in a number of characters, most distinctly in cirrus armature, and from another morphologically similar species, S. sphenomorphus, in the number of proglottids, strobila size, number and size of rostellar hooks, and relative length of cirrus sac. Comparison of partial sequences of nuclear large ribosomal RNA subunit gene (1,310 base pairs [bp]) and mitochondrial NAD(P)H dehydrogenase subunit 1 gene (710 bp) strongly supports the status of the described forms as new species. This increases the number of Staphylocystoides species known in North America to 9 (5 parasitizing S. cinereus) and the total number of species in the genus to 12. Staphylocystoides parasphenomorphus n. sp. is the first named Staphylocystoides record in Canada.

To describe the parasitic community of the Magellanic Horned Owl, Bubo magellanicus (Aves, Strigiformes), 19 carcasses from central Chile were analyzed. Ectoparasites were collected through plumage inspection, while endoparasites were collected through traditional techniques of parasitological necropsy. Sixteen owls were infected with at least one species of ectoparasite (84.21%) or endoparasite (31.58%). Eleven of 19 birds (57.89%) harbored feather mites of the three species Pandalura cirrata (42.11%), Glaucalges attenuatus (47.37%), and Kramerella sp. (10.53%), whereas 16 individuals (84.21%) harbored the chewing louse Strigiphilus chilensis. Only six birds (31.58%) were infected with helminths; the nematodes Capillaria tenuissima (26.32%) and Dispharynx nasuta (5.26%); the acanthocephalan Centrorhynchus spinosus (5.26%); and the trematode Neodiplostomum sp. (5.26%). Apart from S . chilensis, all parasites comprised new records for B . magellanicus.

Cellular uptake of nanoparticles (NPs) depends on the nature of the nanobio system including the solid nanocomponents ( e. g., physicochemical properties of NPs), nanobio interfaces ( e. g., protein corona composition), and the cellular characteristics ( e. g., cell type). In this study, we document the role of sex in cellular uptake of NPs as an "overlooked" factor in nanobio interface investigations. We demonstrate that cell sex leads to differences in NP uptake between male and female human amniotic stem cells (hAMSCs), with greater uptake by female cells. hAMSCs are one of the earliest sources of somatic stem cells. The experiments were replicated with primary fibroblasts isolated from the salivary gland of adult male and female donors of similar ages, and again the extent of NP uptake was altered by cell sex. However, in contrast to hAMSCs, uptake was greater in male cells. We also found out that female versus male amniotic stem cells exhibited different responses to reprogramming into induced pluripotent stem cells (iPSCs) by the Yamanaka factors. Thus, future studies should consider the effect of sex on the nanobio interactions to optimize clinical translation of NPs and iPSC biology and to help researchers to better design and produce safe and efficient therapeutic sex-specific NPs.

To analyze erythropoietin receptor (EpoR) status in tumors, recombinant human erythropoietin (rHuEpo) was labeled with 99m Tc by 99m Tc-centered 1-pot synthesis, resulting in high radiochemical purity, stability, and biological activity. Both in vitro cell culture experiments and biodistribution studies of normal rats demonstrated successful EpoR targeting. The biodistribution of labeled rHuEpo in a NCI-H1975 xenograft model showed tumor accumulation (tumor-to-muscle ratio, 4.27 ± 1.77), confirming the expression of active EpoR in tumors. Thus, as a novel single positron emission computerized tomography tracer for the imaging of EpoR expression in vivo, 99m Tc-rHuEpo is effective for exploring the role of EpoR in cancer growth, metastasis and angiogenesis., (Copyright © 2017 John Wiley & Sons, Ltd.)

Using renewable and biocompatible natural-based resources to construct functional biomaterials has attracted great attention in recent years. In this work, we successfully prepared a series of steroid-based cationic lipids by integrating various steroid skeletons/hydrophobes with ( l -)-arginine headgroups via facile and efficient synthetic approach. The plasmid DNA (pDNA) binding affinity of the steroid-based cationic lipids, average particle sizes, surface potentials, morphologies and stability of the steroid-based cationic lipids/pDNA lipoplexes were disclosed to depend largely on the steroid skeletons. Cellular evaluation results revealed that cytotoxicity and gene transfection efficiency of the steroid-based cationic lipids in H1299 and HeLa cells strongly relied on the steroid hydrophobes. Interestingly, the steroid lipids/pDNA lipoplexes inclined to enter H1299 cells mainly through caveolae and lipid-raft mediated endocytosis pathways, and an intracellular trafficking route of "lipid-raft-mediated endocytosis→lysosome→cell nucleic localization" was accordingly proposed. The study provided possible approach for developing high-performance steroid-based lipid gene carriers, in which the cytotoxicity, gene transfection capability, endocytosis pathways, and intracellular trafficking/localization manners could be tuned/controlled by introducing proper steroid skeletons/hydrophobes. Noteworthy, among the lipids, Cho-Arg showed remarkably high gene transfection efficacy, even under high serum concentration (50% fetal bovine serum), making it an efficient gene transfection agent for practical application., Competing Interests: The authors declare no conflict of interest.