Bone histology supports gregarious behavior and an early ontogenetic stage to Decuriasuchus quartacolonia (Pseudosuchia: Loricata) from the Middle-Late Triassic of Brazil.

Decuriasuchus quartacolonia is a middle-sized basal "rauisuchian" (Pseudosuchia, Loricata) from the Triassic beds of Brazil, whose original description was based on 10 specimens of equivalent size found in aggregation. In this contribution, we explore the osteohistology of its appendicular bones and a rib, aiming to infer growth patterns and ontogeny. In all analyzed bones (except in the rib) from three individuals, we found fibrolamellar bone (FLB) throughout the cortices, a pattern shared with other histologically sampled basal loricatans. We found evidence that suggests an early ontogenetic stage for all studied specimens: absence of secondary osteons, lack of transition from an inner highly vascularized FLB matrix to a poor vascularized parallel-fibred bone, one line of arrested growth, open vascular canals in the external surface of the cortex and absence of an external fundamental system. In addition, we observed that the neurocentral sutures in the caudal to cervical vertebrae of the holotype are unfused, strongly suggesting that these individuals were not skeletally mature, as had been previously assumed. In addition, our data support the prior hypothesis that Decuriasuchus is the oldest-known archosaur to show evidence of gregarious behavior and adds a probable cause of this as a strategy to obtain food and avoid predation until becoming independent as adults. Furthermore, our results open the possibility that Decuriasuchus may represent an earlier growth stage of the larger Prestosuchus chiniquensis, with two specimens recovered from the same stratigraphic level and paleontological site. Since the adult form of Decuriasuchus remains unknown, a deeper anatomical study is needed to discuss the validity of the species. This study case exemplifies the importance of paleohistology as an essential complementary tool to describe a new morphospecies in vertebrate paleontology.

On the dentition, tooth replacement, and taxonomic status of Charruodon tetracuspidatus Abdala & Ribeiro, 2000: A bizarre cynodont from the middle upper Triassic of southern Brazil.

Among the living tetrapods, mammals present a unique tooth replacement pattern, diphyodonty. Therefore, studying the dentition of mammalian ancestors is relevant to a better understanding of how this remarkable feature evolved. However, little is known about the postcanine tooth replacement pattern among Triassic cynodonts. Here, we applied the nondestructive method of microcomputed tomography (microCT) to analyze the dentition of the enigmatic Upper Triassic sectorial-toothed cynodont Charruodon tetracuspidatus (MCP 3934 PV, holotype) from the Candelaria Sequence, Santa Maria Supersequence, Brazil. The microCT-scan data allowed visualization of the replacement dentition and roots of the functional teeth, which provided information to inform interpretations of the ontogenetic stage and taxonomy of the species. A combination of dental and mandibular traits, as well as the small size of the specimen MCP 3934 PV, suggest an early ontogenetic stage. Additionally, the specimen could potentially be an ontogenetically immature form of another taxon, or a yet unknown species of probainognathian cynodont. Therefore, Charruodon tetracuspidatus is here designated as a nomen dubium, given the challenges of maintaining the species as valid.

An eosimiid primate of South Asian affinities in the Paleogene of Western Amazonia and the origin of New World monkeys.

Recent fossil discoveries in Western Amazonia revealed that two distinct anthropoid primate clades of African origin colonized South America near the Eocene/Oligocene transition (ca. 34 Ma). Here, we describe a diminutive fossil primate from Brazilian Amazonia and suggest that, surprisingly, a third clade of anthropoids was involved in the Paleogene colonization of South America by primates. This new taxon, Ashaninkacebus simpsoni gen. et sp. nov., has strong dental affinities with Asian African stem anthropoids: the Eosimiiformes. Morphology-based phylogenetic analyses of early Old World anthropoids and extinct and extant New World monkeys (platyrrhines) support relationships of both Ashaninkacebus and Amamria (late middle Eocene, North Africa) to the South Asian Eosimiidae. Afro-Arabia, then a mega island, played the role of a biogeographic stopover between South Asia and South America for anthropoid primates and hystricognathous rodents. The earliest primates from South America bear little adaptive resemblance to later Oligocene-early Miocene platyrrhine monkeys, and the scarcity of available paleontological data precludes elucidating firmly their affinities with or within Platyrrhini. Nonetheless, these data shed light on some of their life history traits, revealing a particularly small body size and a diet consisting primarily of insects and possibly fruit, which would have increased their chances of survival on a natural floating island during this extraordinary over-water trip to South America from Africa. Divergence-time estimates between Old and New World taxa indicate that the transatlantic dispersal(s) could source in the intense flooding events associated with the late middle Eocene climatic optimum (ca. 40.5 Ma) in Western Africa.

Characterization and Differential Cytotoxicity of Gramicidin Nanoparticles Combined with Cationic Polymer or Lipid Bilayer.

Gramicidin (Gr) nanoparticles (NPs) and poly (diallyl dimethyl ammonium) chloride (PDDA) water dispersions were characterized and evaluated against Gram-positive and Gram-negative bacteria and fungus. Dynamic light scattering for sizing, zeta potential analysis, polydispersity, and colloidal stability over time characterized Gr NPs/PDDA dispersions, and plating and colony-forming units counting determined their microbicidal activity. Cell viabilities of Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans in the presence of the combinations were reduced by 6, 7, and 7 logs, respectively, at 10 µM Gr/10 µg·mL-1 PDDA, 0.5 µM Gr/0. 5µg·mL-1 PDDA, and 0.5 µM Gr/0.5 µg·mL-1 PDDA, respectively. In comparison to individual Gr doses, the combinations reduced doses by half (S. aureus) and a quarter (C. albicans); in comparison to individual PDDA doses, the combinations reduced doses by 6 times (P. aeruginosa) and 10 times (C. albicans). Gr in supported or free cationic lipid bilayers reduced Gr activity against S. aureus due to reduced Gr access to the pathogen. Facile Gr NPs/PDDA disassembly favored access of each agent to the pathogen: PDDA suctioned the pathogen cell wall facilitating Gr insertion in the pathogen cell membrane. Gr NPs/PDDA differential cytotoxicity suggested the possibility of novel systemic uses for the combination.

Histological analysis of ankylothecodonty in Silesauridae (Archosauria: Dinosauriformes) and its implications for the evolution of dinosaur tooth attachment.

Dinosaurs possess a form of tooth attachment wherein an unmineralized periodontal ligament suspends each tooth within a socket, similar to the condition in mammals and crocodylians. However, little information is known about tooth attachment and implantation in their close relatives, the silesaurids. We conducted a histological survey of several silesaurid taxa to determine the nature of tooth attachment in this phylogenetically and paleoecologically important group of archosaurs. Our histological data demonstrate that these early dinosauriforms do not exhibit the crocodilian/dinosaur condition of a permanent gomphosis, nor the rapid ankylosis that is plesiomorphic for amniotes. Instead, all sampled silesaurids exhibit delayed ankylosis, a condition in which teeth pass through a prolonged stage where the teeth are suspended in sockets by a periodontal ligament, followed by eventual mineralization and fusion of the tooth to the jaws. This suggests that tooth attachment in crocodylians and dinosaurs represent the further retention of an early ontogenetic stage compared to silesaurids, a paedomorphic trend that is mirrored in the evolution of synapsid tooth attachment. It also suggests that the dinosaur and crocodylian gomphosis was convergently acquired via heterochrony or, less likely, that the silesaurid condition represents a reversal to a plesiomorphic state. Moreover, if Silesauridae is nested within Ornithischia, a permanent gomphosis could be convergent between the two main dinosaur lineages, Ornithischia and Saurischia. These results demonstrate that dental characters in early archosaur phylogenies must be chosen and defined carefully, taking into account the relative duration of the different phases of dental ontogeny.

Cationic Nanostructures as Adjuvants for Vaccines.

Spherical or discoidal lipid polymer nanostructures bearing cationic charges successfully adsorb a variety of oppositely charged antigens (Ag) such as proteins, peptides, nucleic acids, or oligonucleotides. This report provides instructions for the preparation and physical characterization of four different cationic nanostructures able to combine and deliver antigens to the immune system: (1) dioctadecyl dimethylammonium bromide (DODAB) bilayer fragments (DODAB BF); (2) polystyrene sulfate (PSS) nanoparticles (NPs) covered with one cationic dioctadecyl dimethylammonium bromide bilayer (DODAB) named (PSS/DODAB); (3) cationic NPs of biocompatible polymer poly(methyl methacrylate) (PMMA) prepared by emulsion polymerization of the methyl methacrylate (MMA) monomer in the presence of DODAB BF (PMMA/DODAB NPs); (4) antigen NPs (NPs) where the cationic polymer poly(diallyl dimethyl ammonium chloride) (PDDA) directly combined at nontoxic and low dose with the antigen (Ag); when the oppositely charged model antigen is ovalbumin (OVA), NPs are named PDDA/OVA. These nanostructures provide adequate microenvironments for carrying and delivering antigens to the antigen-presenting cells of the immune system.

Cationic and Biocompatible Polymer/Lipid Nanoparticles as Immunoadjuvants.

Nanostructures have been of paramount importance for developing immunoadjuvants. They must be cationic and non-cytotoxic, easily assembling with usually oppositely charged antigens such as proteins, haptens or nucleic acids for use in vaccines. We obtained optimal hybrid nanoparticles (NPs) from the biocompatible polymer poly(methyl methacrylate) (PMMA) and the cationic lipid dioctadecyl dimethyl ammonium bromide (DODAB) by emulsion polymerization of methyl methacrylate (MMA) in the presence of DODAB. NPs adsorbed ovalbumin (OVA) as a model antigen and we determined their adjuvant properties. Interestingly, they elicited high double immune responses of the cellular and humoral types overcoming the poor biocompatibility of DODAB-based adjuvants of the bilayer type. The results suggested that the novel adjuvant would be possibly of use in a variety of vaccines.

Antimicrobial Polymer-Based Assemblies: A Review.

An antimicrobial supramolecular assembly (ASA) is conspicuous in biomedical applications. Among the alternatives to overcome microbial resistance to antibiotics and drugs, ASAs, including antimicrobial peptides (AMPs) and polymers (APs), provide formulations with optimal antimicrobial activity and acceptable toxicity. AMPs and APs have been delivered by a variety of carriers such as nanoparticles, coatings, multilayers, hydrogels, liposomes, nanodisks, lyotropic lipid phases, nanostructured lipid carriers, etc. They have similar mechanisms of action involving adsorption to the cell wall, penetration across the cell membrane, and microbe lysis. APs, however, offer the advantage of cheap synthetic procedures, chemical stability, and improved adsorption (due to multipoint attachment to microbes), as compared to the expensive synthetic routes, poor yield, and subpar in vivo stability seen in AMPs. We review recent advances in polymer-based antimicrobial assemblies involving AMPs and APs.

An additional brain endocast of the ictidosaur Riograndia guaibensis (Eucynodontia: Probainognathia): intraspecific variation of endocranial traits.

Recently, the morphology and encephalization of the brain endocast of the Triassic non-mammaliaform probainognathian cynodont Riograndia guaibensis were studied. Here, we analyzed the brain endocast of an additional specimen of this species. The new endocast shows well-defined olfactory bulbs and a median sulcus dividing the hemispheres, traits that were not clearly observed in the first studied specimen. Encephalization quotients were also calculated, revealing similar values to other non-mammaliaform cynodonts and lower than those of the first analyzed specimen. The analyzed cranium is slightly larger than the first studied one and may represent an advanced ontogenetic stage. Hence, these differences may be related to the intraspecific variation of this cynodont or alternatively, to the preservation of each specimen.

Biocompatible Lipid Polymer Cationic Nanoparticles for Antigen Presentation.

Biocompatible lipid polymer nanoparticles (NPs) previously used as antimicrobial agents are explored here as immuno-adjuvants. Poly (methyl methacrylate) (PMMA)/dioctadecyldimethylammonium bromide (DODAB)/poly (diallyldimethylammonium chloride) (PDDA) nanoparticles (NPs) were prepared by emulsion polymerization of methyl methacrylate (MMA) in the presence of DODAB and PDDA, with azobisisobutyronitrile (AIBN) as the initiator. NPs characterization after dialysis by dynamic light-scattering yielded 225 ± 2 nm hydrodynamic diameter (Dz), 73 ± 1 mV zeta-potential (ζ), and 0.10 ± 0.01 polydispersity (P). Ovalbumin (OVA) adsorption reduced ζ to 45 ± 2 mV. Balb/c mice immunized with NPs/OVA produced enhanced OVA-specific IgG1 and IgG2a, exhibited moderate delayed type hypersensitivity reaction, and enhanced cytokines production (IL-4, IL-10, IL-2, IFN-γ) by cultured spleen cells. There was no cytotoxicity against cultured macrophages and fibroblasts. Advantages of the PMMA/DODAB/PDDA NPs were high biocompatibility, zeta-potential, colloidal stability, and antigen adsorption. Both humoral and cellular antigen-specific immune responses were obtained.

Supramolecular Nanostructures for Vaccines.

Although this is an era of pandemics and many devastating diseases, this is also a time when bionanotechnology flourishes, illuminating a multidisciplinary field where vaccines are quickly becoming a balsam and a prevention against insidious plagues. In this work, we tried to gain and also give a deeper understanding on nanovaccines and their way of acting to prevent or cure cancer, infectious diseases, and diseases caused by parasites. Major nanoadjuvants and nanovaccines are temptatively exemplified trying to contextualize our own work and its relative importance to the field. The main properties for novel adjuvants seem to be the nanosize, the cationic character, and the biocompatibility, even if it is achieved in a low dose-dependent manner.

Cationic and biocompatible polymer/lipid nanoparticles as immunoadjuvants

Nanostructures have been of paramount importance for developing immunoadjuvants. They must be cationic and non-cytotoxic, easily assembling with usually oppositely charged antigens such as proteins, haptens or nucleic acids for use in vaccines. We obtained optimal hybrid nanoparticles (NPs) from the biocompatible polymer poly(methyl methacrylate) (PMMA) and the cationic lipid dioctadecyl dimethyl ammonium bromide (DODAB) by emulsion polymerization of methyl methacrylate (MMA) in the presence of DODAB. NPs adsorbed ovalbumin (OVA) as a model antigen and we determined their adjuvant properties. Interestingly, they elicited high double immune responses of the cellular and humoral types overcoming the poor biocompatibility of DODAB-based adjuvants of the bilayer type. The results suggested that the novel adjuvant would be possibly of use in a variety of vaccines.

Fatores relacionados à dificuldade de dormir em adultos e idosos cadastrados na Estratégia Saúde da Família / Factors related to difficulty sleeping in adults and older people registered with the Family Health Strategy

OBJETIVO: Identificar os fatores relacionados à dificuldade de dormir em adultos e idosos cadastrados em Unidades Básicas de Saúde com Estratégia Saúde da Família. METODOLOGIA: Trata-se de um estudo transversal e quantitativo, realizado em Três Lagoas (MS), com 300 indivíduos, de ambos os sexos, com idades a partir de 45 anos, cadastrados na Estratégia Saúde da Família do município. Para a coleta de dados foi utilizado um questionário estruturado contendo questões sociodemográficas, de saúde e de rede de apoio social. A variável dependente dificuldade de dormir foi avaliada pela questão: "O(a) senhor(a) tem dificuldade de dormir?". Na análise dos dados, executou-se uma regressão logística binomial. RESULTADOS: A prevalência de dificuldade de dormir foi de 43% (intervalo de confiança de 95% [IC95%] 37 - 49). De acordo com análises ajustadas, foram associados ao desfecho: sexo feminino (Odds Ratio = 2,08; IC95% 1,21 - 3,60), número de medicamentos por dia (Odds Ratio = 1,18; IC95% 1,05 - 1,33), autorrelato de déficit visual (Odds Ratio = 3,56; IC95% 1,54 - 8,25), autorrelato de déficit auditivo (Odds Ratio = 2,28; IC95% 1,18 - 4,41) e isolamento social (Odds Ratio = 2,01; IC95% 1,14 - 3,54). CONCLUSÕES: A definição dos principais fatores relacionados às queixas de insônia é relevante para auxiliar o planejamento dos cuidados e de políticas públicas, com isso visando melhorar a qualidade do sono nos indivíduos em processo de envelhecimento e na velhice.

OBJECTIVE: To identify the factors related to difficulty sleeping in adults and older people registered on the Family Health Strategy at Primary Care Health Centers. METHODS: This is a cross-sectional, quantitative study conducted in the town of Três Lagoas (MS, Brazil), with 300 people of both sexes, aged 45 years and over, registered on the town's Family Health Strategy. Data were collected using a structured questionnaire containing questions on sociodemographics, health, and social support networks. The dependent variable difficulty sleeping was assessed using the question: "Do you have difficulty sleeping?" Data were analyzed with binomial logistic regression. RESULTS: The prevalence of difficulty sleeping was 43% (95% confidence interval [95%CI] 37 - 49). According to the adjusted analyses, the following variables were associated with the outcome: female sex (Odds Ratio = 2.08; 95%CI 1.21 - 3.60); number of medications per day (Odds Ratio = 1.18; 95%CI 1.05 - 1.33); self-report of visual impairment (Odds Ratio = 3.56; 95%CI 1.54 - 8.25); self-report of hearing loss (Odds Ratio = 2.28; 95%CI 1.18 - 4.41); and social isolation (Odds Ratio = 2.01; 95%CI 1.14 - 3.54). CONCLUSIONS: Identification of the principal factors related to complaints of insomnia is of relevance to support planning of care and of public policies, targeting improved sleep quality in people going through the aging process and beyond.

Virtual brain endocast of Antifer (Mammalia: Cervidae), an extinct large cervid from South America.

A diverse fossil record of Cervidae (Mammalia) has been documented in the South American Pleistocene, when these animals arrived during the Great American Biotic Interchange. Using computed tomography-scanning techniques, it is possible to access the endocranial morphology of extinct species. Here, we studied the brain endocast of the extinct late Pleistocene cervid Antifer ensenadensis from southern Brazil, one of the largest forms that lived on this continent, using comparative morphology, geometric morphometrics, and encephalization quotients. The analyzed endocasts demonstrate that A. ensenadensis had a gyrencephalic brain, showing a prominent longitudinal sinus (=sagittal superior sinus), which is also observed in the large South American cervid Blastocerus dichotomus. The encephalization quotient is within the variation of extant cervids, suggesting maintenance of the pattern of encephalization from at least the late Pleistocene. Geometric morphometric analysis suggested a clear and linear allometric trend between brain endocast size and shape, and highlights A. ensenadensis as an extreme form within the analyzed cervids regarding brain morphology.

Cationic Nanostructures for Vaccines Design.

Subunit vaccines rely on adjuvants carrying one or a few molecular antigens from the pathogen in order to guarantee an improved immune response. However, to be effective, the vaccine formulation usually consists of several components: an antigen carrier, the antigen, a stimulator of cellular immunity such as a Toll-like Receptors (TLRs) ligand, and a stimulator of humoral response such as an inflammasome activator. Most antigens are negatively charged and combine well with oppositely charged adjuvants. This explains the paramount importance of studying a variety of cationic supramolecular assemblies aiming at the optimal activity in vivo associated with adjuvant simplicity, positive charge, nanometric size, and colloidal stability. In this review, we discuss the use of several antigen/adjuvant cationic combinations. The discussion involves antigen assembled to 1) cationic lipids, 2) cationic polymers, 3) cationic lipid/polymer nanostructures, and 4) cationic polymer/biocompatible polymer nanostructures. Some of these cationic assemblies revealed good yet poorly explored perspectives as general adjuvants for vaccine design.

Hybrid Nanoparticles of Poly (Methyl Methacrylate) and Antimicrobial Quaternary Ammonium Surfactants.

Quaternary ammonium surfactants (QACs) are microbicides, whereas poly (acrylates) are biocompatible polymers. Here, the physical and antimicrobial properties of two QACs, cetyl trimethyl ammonium bromide (CTAB) or dioctadecyl dimethyl ammonium bromide (DODAB) in poly (methyl methacrylate) (PMMA) nanoparticles (NPs) are compared to those of QACs alone. Methyl methacrylate (MMA) polymerization using DODAB or CTAB as emulsifiers and initiator azobisisobutyronitrile (AIBN) yielded cationic, nanometric, homodisperse, and stable NPs. NPs' physical and antimicrobial properties were assessed from dynamic light scattering (DLS), scanning electron microscopy, and viability curves of Escherichia coli, Staphylococcus aureus, or Candida albicans determined as log(colony-forming unities counting) over a range of [QACs]. NPs were spherical and homodisperse but activity for free QACs was higher than those for QACs in NPs. Inhibition halos against bacteria and yeast were observed only for free or incorporated CTAB in NPs because PMMA/CTAB NPs controlled the CTAB release. DODAB displayed fungicidal activity against C. albicans since DODAB bilayer disks could penetrate the outer glycoproteins fungus layer. The physical properties and stability of the cationic NPs highlighted their potential to combine with other bioactive molecules for further applications in drug and vaccine delivery.

Simple Nanoparticles from the Assembly of Cationic Polymer and Antigen as Immunoadjuvants.

Since antigens are negatively charged, they combine well with positively charged adjuvants. Here, ovalbumin (OVA) (0.1 mg·mL-1) and poly (diallyldimethylammonium chloride) (PDDA) (0.01 mg·mL-1) yielded PDDA/OVA assemblies characterized by dynamic light scattering (DLS) and scanning electron microscopy (SEM) as spherical nanoparticles (NPs) of 170 ± 4 nm hydrodynamic diameter, 30 ± 2 mV of zeta-potential and 0.11 ± 0.01 of polydispersity. Mice immunization with the NPs elicited high OVA-specific IgG1 and low OVA-specific IgG2a production, indicating a Th-2 response. Delayed-type hypersensitivity reaction (DTH) was low and comparable to the one elicited by Al(OH)3/OVA, suggesting again a Th-2 response. PDDA advantages as an adjuvant were simplicity (a single-component adjuvant), low concentration needed (0.01 mg·mL-1 PDDA) combined with antigen yielding neglectable cytotoxicity, and high stability of PDDA/OVA dispersions. The NPs elicited much higher OVA-specific antibodies production than Al(OH)3/OVA. In vivo, the nano-metric size possibly assured antigen presentation by antigen-presenting cells (APC) at the lymph nodes, in contrast to the location of Al(OH)3/OVA microparticles at the site of injection for longer periods with stimulation of local dendritic cells. In the future, it will be interesting to evaluate combinations of the antigen with NPs carrying both PDDA and elicitors of the Th-1 response.