TubeSpin bioreactor 50 for the high-density cultivation of Sf-9 insect cells in suspension

Here we present the TubeSpin bioreactor 50 (TubeSpins) as a simple and disposable culture system for Sf-9 insect cells in suspension. Sf-9 cells had substantially better growth in TubeSpins than in spinner flasks. After inoculation with 106cells/ml, maximal cell densities of 16×106 and 6×106cells/ml were reached in TubeSpins and spinner flasks, respectively. In addition the cell viability in these batch cultures remained above 90% for 10days in TubeSpins but only for 4days in spinner flasks. Inoculation at even higher cell densities reduced the duration of the lag phase. After inoculation at 2.5×106cells/ml, the culture reached the maximum cell density within 3days instead of 7days as observed for inoculation with 106cells/ml. Infection of Sf-9 cells in TubeSpins or spinner flasks with a recombinant baculovirus coding for green fluorescent protein (GFP) resulted in similar GFP-specific fluorescence levels. TubeSpins are thus an attractive option for the small-scale cultivation of Sf-9 cells in suspension and for baculovirus-mediated recombinant protein productio

Convergent evolution of a labile nutritional symbiosis in ants

Ants are among the most successful organisms on Earth. It has been suggested that forming symbioses with nutrient-supplementing microbes may have contributed to their success, by allowing ants to invade otherwise inaccessible niches. However, it is unclear whether ants have evolved symbioses repeatedly to overcome the same nutrient limitations. Here, we address this question by comparing the independently evolved symbioses in Camponotus, Plagiolepis, Formica and Cardiocondyla ants. Our analysis reveals the only metabolic function consistently retained in all of the symbiont genomes is the capacity to synthesise tyrosine. We also show that in certain multi-queen lineages that have co-diversified with their symbiont for millions of years, only a fraction of queens carry the symbiont, suggesting ants differ in their colony-level reliance on symbiont-derived resources. Our results imply that symbioses can arise to solve common problems, but hosts may differ in their dependence on symbionts, highlighting the evolutionary forces influencing the persistence of long-term endosymbiotic mutualisms.Peer reviewe

TubeSpin bioreactor 50 for the high-density cultivation of Sf-9 insect cells in suspension

Here we present the TubeSpin bioreactor 50 (TubeSpins) as a simple and disposable culture system for Sf-9 insect cells in suspension. Sf-9 cells had substantially better growth in TubeSpins than in spinner flasks. After inoculation with 106 cells/ml, maximal cell densities of 16 × 106 and 6 × 106 cells/ml were reached in TubeSpins and spinner flasks, respectively. In addition the cell viability in these batch cultures remained above 90% for 10 days in TubeSpins but only for 4 days in spinner flasks. Inoculation at even higher cell densities reduced the duration of the lag phase. After inoculation at 2.5 × 106 cells/ml, the culture reached the maximum cell density within 3 days instead of 7 days as observed for inoculation with 106 cells/ml. Infection of Sf-9 cells in TubeSpins or spinner flasks with a recombinant baculovirus coding for green fluorescent protein (GFP) resulted in similar GFP-specific fluorescence levels. TubeSpins are thus an attractive option for the small-scale cultivation of Sf-9 cells in suspension and for baculovirus-mediated recombinant protein production

Angular correlation of the two gamma rays produced in the thermal neutron capture on gadolinium-155 and gadolinium-157

The ANNRI-Gd collaboration studied in detail the single $\gamma$-ray spectrum produced from the thermal neutron capture on $^{155}$Gd and $^{157}$Gd in our previous publications. Gadolinium targets were exposed to a neutron beam provided by the Japan Spallation Neutron Source (JSNS) in J-PARC, Japan. In the present analysis, one new additional coaxial germanium crystal was used in the analysis in combination with the fourteen germanium crystals in the cluster detectors to study the angular correlation of the two $\gamma$ rays emitted in the same neutron capture. We present for the first time angular correlation functions for two $\gamma$ rays produced during the electromagnetic cascade transitions in the (n, $\gamma$) reactions on $^{\rm 155}$Gd and $^{\rm 157}$Gd. As expected, we observe the mild angular correlations for the strong, but rare transitions from the resonance state to the two energy levels of known spin-parities. Contrariwise, we observe negligibly small angular correlations for arbitrary pairs of two $\gamma$ rays produced in the majority of cascade transitions from the resonance state to the dense continuum states.Comment: 21 pages, 13 figure

Gamma Ray Spectra from Thermal Neutron Capture on Gadolinium-155 and Natural Gadolinium

Abstract Natural gadolinium is widely used for its excellent thermal neutron capture cross section, because of its two major isotopes: $^{\rm 155}$Gd and $^{\rm 157}$Gd. We measured the $\gamma$-ray spectra produced from the thermal neutron capture on targets comprising a natural gadolinium film and enriched $^{\rm 155}$Gd (in Gd$_{2}$O$_{3}$ powder) in the energy range from 0.11 MeV to 8.0 MeV, using the ANNRI germanium spectrometer at MLF, J-PARC. The freshly analyzed data of the $^{\rm 155}$Gd($n, \gamma$) reaction are used to improve our previously developed model (ANNRI-Gd model) for the $^{\rm 157}$Gd($n, \gamma$) reaction [K. Hagiwara et al. [ANNRI-Gd Collaboration], Prog. Theor. Exp. Phys. 2019, 023D01 (2019)], and its performance confirmed with the independent data from the $^{\rm nat}$Gd($n, \gamma$) reaction. This article completes the development of an efficient Monte Carlo model required to simulate and analyze particle interactions involving the thermal neutron captures on gadolinium in any relevant future experiments

Masses, radii, and orbits of small Kepler planets : The transition from gaseous to rocky planets

We report on the masses, sizes, and orbits of the planets orbiting 22 Kepler stars. There are 49 planet candidates around these stars, including 42 detected through transits and 7 revealed by precise Doppler measurements of the host stars. Based on an analysis of the Kepler brightness measurements, along with high-resolution imaging and spectroscopy, Doppler spectroscopy, and (for 11 stars) asteroseismology, we establish low false-positive probabilities (FPPs) for all of the transiting planets (41 of 42 have an FPP under 1%), and we constrain their sizes and masses. Most of the transiting planets are smaller than three times the size of Earth. For 16 planets, the Doppler signal was securely detected, providing a direct measurement of the planet's mass. For the other 26 planets we provide either marginal mass measurements or upper limits to their masses and densities; in many cases we can rule out a rocky composition. We identify six planets with densities above 5 g cm-3, suggesting a mostly rocky interior for them. Indeed, the only planets that are compatible with a purely rocky composition are smaller than 2 R ⊕. Larger planets evidently contain a larger fraction of low-density material (H, He, and H2O).Peer reviewedFinal Accepted Versio

Changing word usage predicts changing word durations in New Zealand English

This paper investigates the emergence of lexicalized effects of word usage on word duration by looking at parallel changes in usage and duration over 130 years in New Zealand English. Previous research has found that frequent words are shorter, informative words are longer, and words in utterance-final position are also longer. It has also been argued that some of these patterns are not simply online adjustments, but are incorporated into lexical representations. While these studies tend to focus on the synchronic aspects of such patterns, our corpus shows that word-usage patterns and word durations are not static over time. Many words change in duration and also change with respect to frequency, informativity and likelihood of occurring utterance-finally. Analysis of changing word durations over this time period shows substantial patterns of co-adaptation between word usage and word durations. Words that are increasing in frequency are becoming shorter. Words that are increasing/decreasing in informativity show a change in the same direction in duration (e.g. increasing informativity is associated with increasing duration). And words that are increasingly appearing utterance-finally are lengthening. These effects exist independently of the local effects of the predictors. For example, words that are increasing utterance-finally lengthen in all positions, including utterance-medially. We show that these results are compatible with a number of different views about lexical representations, but they cannot be explained without reference to a production-perception loop that allows speakers to update their representations dynamically on the basis of their experience

Functional Implication of Dp71 in Osmoregulation and Vascular Permeability of the Retina

Functional alterations of Müller cells, the principal glia of the retina, are an early hallmark of most retina diseases and contribute to their further progression. The molecular mechanisms of these reactive Müller cell alterations, resulting in disturbed retinal homeostasis, remain largely unknown. Here we show that experimental detachment of mouse retina induces mislocation of the inwardly rectifying potassium channels (Kir4.1) and a downregulation of the water channel protein (AQP4) in Müller cells. These alterations are associated with a strong decrease of Dp71, a cytoskeleton protein responsible for the localization and the clustering of Kir4.1 and AQP4. Partial (in detached retinas) or total depletion of Dp71 in Müller cells (in Dp71-null mice) impairs the capability of volume regulation of Müller cells under osmotic stress. The abnormal swelling of Müller cells In Dp71-null mice involves the action of inflammatory mediators. Moreover, we investigated whether the alterations in Müller cells of Dp71-null mice may interfere with their regulatory effect on the blood-retina barrier. In the absence of Dp71, the retinal vascular permeability was increased as compared to the controls. Our results reveal that Dp71 is crucially implicated in the maintenance of potassium homeostasis, in transmembraneous water transport, and in the Müller cell-mediated regulation of retinal vascular permeability. Furthermore, our data provide novel insights into the mechanisms of retinal homeostasis provided by Müller cells under normal and pathological conditions