Gastrulation in Halocordyle disticha (Hydrozoa,Athecata)

Summary

Previous reports of development in Halocordyle disticha have described gastrulation as occurring by gradual differentiation of the inner and outer cells of the stereoblastula. In 1984, however, Martin and Thomas described an indentation on the surface of the embryo at the time of gastrulation. They hypothesized from morphological data that the indentation represented a blastopore. Here we provide results of marking studies which demonstrate that the indentation is in fact a site of cellular ingression. This is the first example known of gastrulation that involves unipolar ingression in a form with a stereoblastula. Possible functions of gastrulation by unipolar ingression are discussed, and the possible phylogenetic significance of the occurrence of such a mode of gastrulation in H. disticha is considered.     

Comprehensive analysis of human immunodeficiency virus type 1-specific CD4 responses reveals marked immunodominance of gag and nef and the presence of broadly recognized peptides

Increasing evidence suggests that human immunodeficiency virus type 1 (HIV-1)-specific CD4 T-cell responses contribute to effective immune control of HIV-1 infection. However, the breadths and specificities of these responses have not been defined. We screened fresh CD8-depleted peripheral blood mononuclear cells (PBMC) from 36 subjects at different stages of HIV-1 infection for virus-specific CD4 responses by gamma interferon enzyme-linked immunospot assay, using 410 overlapping peptides spanning all HIV-1 proteins (based on the clade B consensus sequence). HIV-1-specific CD4 responses were identified in 30 of the 36 individuals studied, with the strongest and broadest responses detected in persons treated in acute infection who underwent treatment interruption. In individuals with identified responses, the total number of recognized HIV-1 peptides ranged from 1 to 36 (median, 7) and the total magnitude of responses ranged from 80 to >14,600 (median, 990) spot-forming cells/10(6) CD8-depleted PBMC. Neither the total magnitude nor the number of responses correlated with viremia. The most frequent and robust responses were directed against epitopes within the Gag and Nef proteins. Peptides targeted by >/=25% of individuals were then tested for binding to a panel of common HLA-DR molecules. All bound broadly to at least four of the eight alleles tested, and two bound to all of the HLA-DR molecules studied. Fine mapping and HLA restriction of the responses against four of these peptides showed a combination of clustering of epitopes and promiscuous presentation of the same epitopes by different HLA class II alleles. These findings have implications for the design of immunotherapeutic strategies and for testing candidate HIV vaccines.     

Widespread gene flow between oceans in a pelagic seabird species complex

Global‐scale gene flow is an important concern in conservation biology as it has the potential to either increase or decrease genetic diversity in species and populations. Although many studies focus on the gene flow between different populations of a single species, the potential for gene flow and introgression between species is understudied, particularly in seabirds. The only well‐studied example of a mixed‐species, hybridizing population of petrels exists on Round Island, in the Indian Ocean. Previous research assumed that Round Island represents a point of secondary contact between Atlantic (Pterodroma arminjoniana) and Pacific species (Pterodroma neglecta and Pterodroma heraldica). This study uses microsatellite genotyping and tracking data to address the possibility of between‐species hybridization occurring outside the Indian Ocean. Dispersal and gene flow spanning three oceans were demonstrated between the species in this complex. Analysis of migration rates estimated using bayesass revealed unidirectional movement of petrels from the Atlantic and Pacific into the Indian Ocean. Conversely, structure analysis revealed gene flow between species of the Atlantic and Pacific oceans, with potential three‐way hybrids occurring outside the Indian Ocean. Additionally, geolocation tracking of Round Island petrels revealed two individuals travelling to the Atlantic and Pacific. These results suggest that interspecific hybrids in Pterodroma petrels are more common than was previously assumed. This study is the first of its kind to investigate gene flow between populations of closely related Procellariiform species on a global scale, demonstrating the need for consideration of widespread migration and hybridization in the conservation of threatened seabirds.     

Evaluation of piggyBac‐mediated CHO pools to enable material generation to support GLP toxicology studies

Generating purified protein for GLP toxicology studies (GLP‐Tox) represents an important and often rate limiting step in the biopharmaceutical drug development process. Toxicity testing requires large amounts of therapeutic protein (>100 g), typically produced in a single 500–2,500 L bioreactor, using the final CHO clonally derived cell line (CDCL). One approach currently used to save time is to manufacture GLP‐Tox material using pools of high‐producing CHO CDCLs instead of waiting for the final CDCL. Recently, we reported CHO pools producing mAb titers >7 g/L using piggyBac‐mediated gene integration (PB CHO pools). In this study, we wanted to leverage high titer PB CHO pools to produce GLP‐Tox material. A detailed product quality attribute (PQA) assessment was conducted comparing PB CHO pools to pooled Top4 CDCLs. Four mAbs were evaluated. First, we found that PB CHO pools expressed all four mAbs at high titers (2.8–4.4 g/L in shake flasks). Second, all four PB CHO pools were aged to 55 generations (Gen). All four PB CHO Pools were found to be suitable over 55 Gen. Finally, we performed bioreactor scale‐up. PB CHO pool titers (3.7–4.8 g/L) were similar or higher than the pooled Top 4 CDCLs in 5 L bioreactors (2.4–4.1 g/L). The PQAs of protein derived from PB CHO pools were very similar to pooled Top 4 CHO CDCLs according to multiple orthogonal techniques including peptide mapping analysis. Taken together, these results demonstrate the technical feasibility of using PB CHO pools to manufacture protein for GLP‐Tox. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1436–1448, 2017     

Sodium arsenite affects Na+ transport in the isolated skin of the toad Pleurodema thaul

Arsenic, applied as sodium arsenite (As(III)) to either inner or outer surfaces of the isolated toad skin, dose-dependently decreased the short-circuit current (Isc), potential difference (PD) and sodium conductance (G(Na)) in the concentration range 1-1000 microM, with effects often lasting over 3 h. Maximal inhibitory effect was over 90% with an IC(50) of about 34 microM. Applied during amiloride block, As(III) did not change this effect. However, an increase in electric parameters was noted during the initial 30 min in 22 experiments, indicating a possible translocation of cytosolic protein kinase C (PKC) to the membrane within 15 min, thus stimulating sodium transport; this is followed by a progressive inhibition of kinase activity. Comparative effects of amiloride (8 microM), As(III) (100 microM, outer surface) and noradrenaline (NA, 10 microM, inner surface) showed a significant increase in the stimulatory effect of NA on the electric parameters, which could be the result of arsenite clustering of cell surface receptors and activation of ensuing cellular signal transduction pathways. Ouabain 5 microM, followed by As(III) 100 microM, also stimulated the skin response to NA (10 microM), although the duration of the two phases of the response was markedly shortened. The exact mechanism is still in doubt: however, As(III) increases cerebral metabolites of NA and ouabain can increase NA efflux from tissue slices. The amiloride test, performed with As(III) in the outer surface, confirmed significant decrease in all the parameters: the driving force (E(Na)), sodium conductance (G(Na)), and importantly, shunt conductance (G(sh)), due to the known fact that arsenic inhibits gap junctional intercellular communication.     

Does male extra‐territory foray effort affect fertilization success in hooded warblers Wilsonia citrina?

Hooded warbler Wilsonia citrina males vary greatly in the frequency and duration of their off-territory forays in search of extra-pair copulations. We used radiotracking and microsatellite parentage analysis in high and low density populations to determine if (1) high foray rate or time off-territory reduces within-pair fertilization success, and (2) if a high foray rate onto the territory of a fertile female increases the likelihood of obtaining EPFs with that female. Males who left their territory often, or for longer periods, did not have lower within-pair fertilization success. Some males repeatedly visited a neighboring fertile female, but in only 3 of 19 cases where radiotagged males visited a fertile female did the male actually sire offspring with that female. Male foray rate onto a fertile female's territory was not a good predictor of whether or not he sired extra-pair offspring with that female. We suggest that mate choice and extra-pair behavior by females may explain why male foray behavior does not correspond closely with actual fertilization success.     

A general modeling framework for describing spatially structured population dynamics

Variation in movement across time and space fundamentally shapes the abundance and distribution of populations. Although a variety of approaches model structured population dynamics, they are limited to specific types of spatially structured populations and lack a unifying framework. Here, we propose a unified network‐based framework sufficiently novel in its flexibility to capture a wide variety of spatiotemporal processes including metapopulations and a range of migratory patterns. It can accommodate different kinds of age structures, forms of population growth, dispersal, nomadism and migration, and alternative life‐history strategies. Our objective was to link three general elements common to all spatially structured populations (space, time and movement) under a single mathematical framework. To do this, we adopt a network modeling approach. The spatial structure of a population is represented by a weighted and directed network. Each node and each edge has a set of attributes which vary through time. The dynamics of our network‐based population is modeled with discrete time steps. Using both theoretical and real‐world examples, we show how common elements recur across species with disparate movement strategies and how they can be combined under a unified mathematical framework. We illustrate how metapopulations, various migratory patterns, and nomadism can be represented with this modeling approach. We also apply our network‐based framework to four organisms spanning a wide range of life histories, movement patterns, and carrying capacities. General computer code to implement our framework is provided, which can be applied to almost any spatially structured population. This framework contributes to our theoretical understanding of population dynamics and has practical management applications, including understanding the impact of perturbations on population size, distribution, and movement patterns. By working within a common framework, there is less chance that comparative analyses are colored by model details rather than general principles.     

Convergent evolution of floral signals underlies the success of Neotropical orchids

The great majority of plant species in the tropics require animals to achieve pollination, but the exact role of floral signals in attraction of animal pollinators is often debated. Many plants provide a floral reward to attract a guild of pollinators, and it has been proposed that floral signals of non-rewarding species may converge on those of rewarding species to exploit the relationship of the latter with their pollinators. In the orchid family (Orchidaceae), pollination is almost universally animal-mediated, but a third of species provide no floral reward, which suggests that deceptive pollination mechanisms are prevalent. Here, we examine floral colour and shape convergence in Neotropical plant communities, focusing on certain food-deceptive Oncidiinae orchids (e.g. Trichocentrum ascendens and Oncidium nebulosum) and rewarding species of Malpighiaceae. We show that the species from these two distantly related families are often more similar in floral colour and shape than expected by chance and propose that a system of multifarious floral mimicry—a form of Batesian mimicry that involves multiple models and is more complex than a simple one model–one mimic system—operates in these orchids. The same mimetic pollination system has evolved at least 14 times within the species-rich Oncidiinae throughout the Neotropics. These results help explain the extraordinary diversification of Neotropical orchids and highlight the complexity of plant–animal interactions.