<Emphasis Type="Italic">Megaviridae</Emphasis>-like particles associated with <Emphasis Type="Italic">Symbiodinium</Emphasis> spp. from the endemic coral <Emphasis Type="Italic">Mussismilia braziliensis</Emphasis>

Coral reefs are one of the most dynamic and productive marine ecosystems. The coral holobiont consists of the coral animal and a variety of associated microorganisms that include symbiotic dinoflagellates of the genus Symbiodinium, bacteria, archaea, fungi and viruses. The interactions among these components are crucial for coral health and, consequently, to the coral reef resilience to disturbance. Environmental stressors such as elevated temperature, high irradiance and ultraviolet (UV) radiation can lead to the breakdown of the coral-Symbiodinium symbiosis in a phenomenon known as “coral bleaching”. The present study provides evidence for virus-like particles (VLPs) induced in UV-irradiated Symbiodinium spp. cultures (clades A and C) that were isolated from the coral Mussismilia braziliensis, suggesting a latent viral infection in these strains. Scanning and transmission electron microscopy images of the UV stressed cultures revealed the presence of giant (ca. 450 nm) and small (ca. 40 nm) VLPs. Morphological features link the giant VLPs to the family Megaviridae. Symbiodinium spp. Megaviridae giant viruses and other associated viruses may represent dynamic forces driving and influencing health of the coral holobiont.     

Potential for colonization of O111:H25 atypical enteropathogenic <Emphasis Type="Italic">E. coli</Emphasis>

Using clonal phylogenetic methods, it has been demonstrated that O111:H25 atypical enteropathogenic E. coli (aEPEC) strains belong to distinct clones, suggesting the possibility that their ability to interact with different hosts and abiotic surfaces can vary from one clone to another. Accordingly, the ability of O111:H25 aEPEC strains derived from human, cat and dogs to adhere to epithelial cells has been investigated, along with their ability to interact with macrophages and to form biofilms on polystyrene, a polymer used to make biomedical devices. The results demonstrated that all the strains analyzed were able to adhere to, and to form pedestals on, epithelial cells, mechanisms used by E. coli to become strongly attached to the host. The strains also show a Localized-Adherence-Like (LAL) pattern of adhesion on HEp-2 cells, a behavior associated with acute infantile diarrhea. In addition, the O111:H25 aEPEC strains derived either from human or domestic animals were able to form long filaments, a phenomenon used by some bacteria to avoid phagocytosis. O111:H25 aEPEC strains were also encountered inside vacuoles, a characteristic described for several bacterial strains as a way of protecting themselves against the environment. They were also able to induce TNF-α release via two routes, one dependent on TLR-4 and the other dependent on binding of Type I fimbriae. These O111:H25 strains were also able to form biofilms on polystyrene. In summary the results suggest that, regardless of their source (i.e. linked to human origin or otherwise), O111:H25 aEPEC strains carry the potential to cause human disease.     

Environmental Mapping of Paracoccidioides spp. in Brazil Reveals New Clues into Genetic Diversity,Biogeography and Wild Host Association

BackgroundParacoccidioides brasiliensis and Paracoccidioides lutzii are the etiological agents of Paracoccidioidomycosis (PCM), and are easily isolated from human patients. However, due to human migration and a long latency period, clinical isolates do not reflect the spatial distribution of these pathogens. Molecular detection of P. brasiliensis and P. lutzii from soil, as well as their isolation from wild animals such as armadillos, are important for monitoring their environmental and geographical distribution. This study aimed to detect and, for the first time, evaluate the genetic diversity of P. brasiliensis and P. lutzii for Paracoccidioidomycosis in endemic and non-endemic areas of the environment, by using Nested PCR and in situ hybridization techniques.Conclusions/SignificanceData may reflect the actual occurrence of Paracoccidioides species in their saprobic habitat, despite their absence/non-detection in seven armadillos evaluated in regions with high prevalence of PCM infection by P. lutzii. These results may indicate a possible ecological difference between P. brasiliensis and P. lutzii concerning their wild hosts.     

An efficient route to bispecific antibody production using single-reactor mammalian co-culture

Bispecific antibodies have shown promise in the clinic as medicines with novel mechanisms of action. Lack of efficient production of bispecific IgGs, however, has limited their rapid advancement. Here, we describe a single-reactor process using mammalian cell co-culture production to efficiently produce a bispecific IgG with 4 distinct polypeptide chains without the need for parallel processing of each half-antibody or additional framework mutations. This method resembles a conventional process, and the quality and yield of the monoclonal antibodies are equal to those produced using parallel processing methods. We demonstrate the application of the approach to diverse bispecific antibodies, and its suitability for production of a tissue specific molecule targeting fibroblast growth factor receptor 1 and klotho β that is being developed for type 2 diabetes and other obesity-linked disorders.     

Oral insulin delivery by means of solid lipid nanoparticles

The aim of this work was to produce and characterize cetyl palmitate-based solid lipid nanoparticles (SLN) containing insulin, and to evaluate the potential of these colloidal carriers for oral administration. SLN were prepared by a modified solvent emulsification-evaporation method based on a w/o/w double emulsion. The particle size, zeta potential and association efficiency of unloaded and insulin-loaded SLN were determined and were found to be around 350 nm, negatively charged and the insulin association efficiency was over 43%. After oral administration of insulin-loaded SLN to diabetic rats, a considerable hypoglycemic effect was observed during 24 hours. These results demonstrated that SLN promote the oral absorption of insulin.     

The Effects of Different Training Backgrounds on VO2 Responses to All-Out and Supramaximal Constant-Velocity Running Bouts

To investigate the impact of different training backgrounds on pulmonary oxygen uptake (V̇O₂) responses during all-out and supramaximal constant-velocity running exercises, nine sprinters (SPRs) and eight endurance runners (ENDs) performed an incremental test for maximal aerobic velocity (MAV) assessment and two supramaximal running exercises (1-min all-out test and constant-velocity exercise). The V̇O₂ responses were continuously determined during the tests (K4^b2, Cosmed, Italy). A mono-exponential function was used to describe the V̇O₂ onset kinetics during constant-velocity test at 110%MAV, while during 1-min all-out test the peak of V̇O₂ (V̇O₂peak), the time to achieve the V̇O₂peak (tV̇O₂peak) and the V̇O₂ decrease at last of the test was determined to characterize the V̇O₂ response. During constant-velocity exercise, ENDs had a faster V̇O₂ kinetics than SPRs (12.7 ± 3.0 vs. 19.3 ± 5.6 s; p < 0.001). During the 1-min all-out test, ENDs presented slower tV̇O₂peak than SPRs (40.6 ± 6.8 and 28.8 ± 6.4 s, respectively; p = 0.002) and had a similar V̇O₂peak relative to the V̇O₂max (88 ± 8 and 83 ± 6%, respectively; p = 0.157). Finally, SPRs was the only group that presented a V̇O₂ decrease in the last half of the test (-1.8 ± 2.3 and 3.5 ± 2.3 ml.kg^-1.min^-1, respectively; p < 0.001). In summary, SPRs have a faster V̇O₂ response when maximum intensity is required and a high maximum intensity during all-out running exercise seems to lead to a higher decrease in V̇O₂ in the last part of the exercise.     

Description of Euroleptochromus gen.n. (Coleoptera,Staphylinidae, Scydmaeninae) from Baltic amber,with discussion of biogeography and mouthpart evolution within Clidicini

A new extinct species of the ant‐like stone beetle supertribe Mastigitae, Euroleptochromus sabathi gen. & sp.n. is described from Eocene Baltic amber. A phylogenetic analysis of Clidicini, with representatives of Leptomastacini and Mastigini as out‐group taxa, provided strong support for a sister‐group relationship between the Neotropical Leptochromus and the new genus. The monophyly of Clidicini is questioned because of an alternative placement of Nearctic Papusus as a sister taxon to Leptomastacini + [Clidicus + (Palaeoleptochromus + (Euroleptochromus + Leptochromus))]. A dispersal‐vicariance analysis provided three alternative scenarios for the evolution of Mastigitae; with Laurasia as the ancestral area of the supertribe, major branching events occurring within either Eurasia or Laurentia and two trans‐Beringia dispersals in Late Cretaceous and Eocene. Euroleptochromus, Palaeoleptochromus and Leptochromus share highly derived structures on postgenae and maxillary palps, probably as part of a specialised feeding or prey capture mechanism. The formation of these modifications in Clidicini is demonstrated to involve a process (traced back to the Campanian, 79 Ma) of elongation and narrowing of maxillary palps and forming a cuticular setal projection from a broadened insertion site of sensory setae.