Dwarf Novae in the Shortest Orbital Period Regime: I. A New Short Period Dwarf Nova, OT J055717+683226

We report the observation of a new dwarf nova, OT J055717+683226, during its first-ever recorded superoutburst in December 2006. Our observation shows that this object is an SU UMa-type dwarf nova having a very short superhump period of 76.67+/- 0.03 min (0.05324+/-0.00002 d). The next superoutburst was observed in March 2008. The recurrence time of superoutbursts (supercycle) is, hence, estimated to be ~480 d. The supercycle is much shorter than those of WZ Sge-type dwarf novae having supercycles of >~ 10 yr, which are a major population of dwarf novae in the shortest orbital period regime (<~85 min). Using a hierarchical cluster analysis, we identified seven groups of dwarf novae in the shortest orbital period regime. We identified a small group of objects that have short supercycles, small outburst amplitudes, and large superhump period excesses, compared with those of WZ Sge stars. OT J055717+683226 probably belongs to this group.Comment: 14 pages, 11 figures, accepted for publication in PAS

Moving toward generalizable NZ-1 labeling for 3D structure determination with optimized epitope-tag insertion

タンパク質の抗体ラベリング技術を改良し、構造解析をアシスト --電子顕微鏡やX線結晶解析による構造決定を加速化--. 京都大学プレスリリース. 2021-04-20.Antibody labeling has been conducted extensively for structure determination using both X-ray crystallography and electron microscopy (EM). However, establishing target-specific antibodies is a prerequisite for applying antibody-assisted structural analysis. To expand the applicability of this strategy, an alternative method has been developed to prepare an antibody complex by inserting an exogenous epitope into the target. It has already been demonstrated that the Fab of the NZ-1 monoclonal antibody can form a stable complex with a target containing a PA12 tag as an inserted epitope. Nevertheless, it was also found that complex formation through the inserted PA12 tag inevitably caused structural changes around the insertion site on the target. Here, an attempt was made to improve the tag-insertion method, and it was consequently discovered that an alternate tag (PA14) could replace various loops on the target without inducing large structural changes. Crystallographic analysis demonstrated that the inserted PA14 tag adopts a loop-like conformation with closed ends in the antigen-binding pocket of the NZ-1 Fab. Due to proximity of the termini in the bound conformation, the more optimal PA14 tag had only a minor impact on the target structure. In fact, the PA14 tag could also be inserted into a sterically hindered loop for labeling. Molecular-dynamics simulations also showed a rigid structure for the target regardless of PA14 insertion and complex formation with the NZ-1 Fab. Using this improved labeling technique, negative-stain EM was performed on a bacterial site-2 protease, which enabled an approximation of the domain arrangement based on the docking mode of the NZ-1 Fab

Application of unfolding transformation in the random matrix theory to analyze in vivo neuronal spike firing during awake and anesthetized conditions

General anesthetics decrease the frequency and density of spike firing. This effect makes it difficult to detect spike regularity. To overcome this problem, we developed a method utilizing the unfolding transformation which analyzes the energy level statistics in the random matrix theory. We regarded the energy axis as time axis of neuron spike and analyzed the time series of cortical neural firing in vivo. Unfolding transformation detected regularities of neural firing while changes in firing densities were associated with pentobarbital. We found that unfolding transformation enables us to compare firing regularity between awake and anesthetic conditions on a universal scale. Keywords: Unfolding transformation, Spike-timing, Regularit

Spike timing rigidity is maintained in bursting neurons under pentobarbital-induced anesthetic conditions

Pentobarbital potentiates γ-aminobutyric acid (GABA)-mediated inhibitory synaptic transmission by prolonging the open time of GABAA receptors. However, it is unknown how pentobarbital regulates cortical neuronal activities via local circuits in vivo. To examine this question, we performed extracellular unit recording in rat insular cortex under awake and anesthetic conditions. Not a few studies apply time-rescaling theorem to detect the features of repetitive spike firing. Similar to these methods, we define an average spike interval locally in time using random matrix theory (RMT), which enables us to compare different activity states on a universal scale. Neurons with high spontaneous firing frequency (> 5 Hz) and bursting were classified as HFB neurons (n = 10), and those with low spontaneous firing frequency (< 10 Hz) and without bursting were classified as non-HFB neurons (n = 48). Pentobarbital injection (30 mg/kg) reduced firing frequency in all HFB neurons and in 78% of non-HFB neurons. RMT analysis demonstrated that pentobarbital increased in the number of neurons with repulsion in both HFB and non-HFB neurons, suggesting that there is a correlation between spikes within a short interspike interval. Under awake conditions, in 50% of HFB and 40% of non-HFB neurons, the decay phase of normalized histograms of spontaneous firing were fitted to an exponential function, which indicated that the first spike had no correlation with subsequent spikes. In contrast, under pentobarbital-induced anesthesia conditions, the number of non-HFB neurons that were fitted to an exponential function increased to 80%, but almost no change in HFB neurons was observed. These results suggest that under both awake and pentobarbital-induced anesthetized conditions, spike firing in HFB neurons is more robustly regulated by preceding spikes than by non-HFB neurons, which may reflect the GABAA receptor-mediated regulation of cortical activities. Whole-cell patch-clamp recording in the IC slice preparation was performed to compare the regularity of spike timing between pyramidal and fast-spiking (FS) neurons, which presumably correspond to non-HFB and HFB neurons, respectively. Repetitive spike firing of FS neurons exhibited a lower variance of interspike interval than pyramidal neurons both in control and under application of pentobarbital, supporting the above hypothesis

Influence of Rice Husk Biochar on Soil Nematode Community under Upland and Flooded Conditions: A Microcosm Experiment

Biochar has the potential for improving soil properties and supporting ecological functions, but it has negative impacts on soil organisms in some cases. This study aimed to assess the effect of biochar application at rates of 0 (B0), 5 Mg ha&minus;1 (B5), 20 Mg&minus;1 (B20), and 40 Mg ha&minus;1 (B40) on soil nematode community under upland and flooded conditions in a short-term microcosm experiment. After biochar application, soil was incubated for 2 to 8 weeks and nematodes were identified for community composition, trophic structures, functional guilds, maturity index and metabolic footprints. The chemical properties of the soils were also analyzed. General linear model revealed that biochar increased soil pH, EC, NO3&minus;-N, available phosphorus, total C, and C/N ratio, particularly in the highest application rate and shifted the composition of nematodes. The greatest abundances of omnivores (Mesodorylaimus, Thornenema), predator (Nygolaimus) and functional guilds of cp5 were observed in B5, resulting in greatest structure footprint and composite footprint, (omnivorous + predator) footprint and total biomass in B5. While abundances of nematodes tended to decrease with the biochar applicate rates, the abundance of Prismatolaimus was the highest in B40. During the 8-week incubation period, the abundances of Achromadora, Alaimus, Aporcelaimellus, Cryptonchus, Mononchus, and Tobrilus remained stable in upland conditions. Under flooded conditions, the abundances of almost all taxa were markedly lower than those under upland conditions irrespective of biochar application, except for Acrobeloides, Alaimus, Aphelenchoides, and Ditylenchus. We highlighted that 5 Mg ha&minus;1 of rice husk biochar can be the optimum in shaping the nematode community

Influence of Rice Husk Biochar on Soil Nematode Community under Upland and Flooded Conditions: A Microcosm Experiment

Biochar has the potential for improving soil properties and supporting ecological functions, but it has negative impacts on soil organisms in some cases. This study aimed to assess the effect of biochar application at rates of 0 (B0), 5 Mg ha−1 (B5), 20 Mg−1 (B20), and 40 Mg ha−1 (B40) on soil nematode community under upland and flooded conditions in a short-term microcosm experiment. After biochar application, soil was incubated for 2 to 8 weeks and nematodes were identified for community composition, trophic structures, functional guilds, maturity index and metabolic footprints. The chemical properties of the soils were also analyzed. General linear model revealed that biochar increased soil pH, EC, NO3−-N, available phosphorus, total C, and C/N ratio, particularly in the highest application rate and shifted the composition of nematodes. The greatest abundances of omnivores (Mesodorylaimus, Thornenema), predator (Nygolaimus) and functional guilds of cp5 were observed in B5, resulting in greatest structure footprint and composite footprint, (omnivorous + predator) footprint and total biomass in B5. While abundances of nematodes tended to decrease with the biochar applicate rates, the abundance of Prismatolaimus was the highest in B40. During the 8-week incubation period, the abundances of Achromadora, Alaimus, Aporcelaimellus, Cryptonchus, Mononchus, and Tobrilus remained stable in upland conditions. Under flooded conditions, the abundances of almost all taxa were markedly lower than those under upland conditions irrespective of biochar application, except for Acrobeloides, Alaimus, Aphelenchoides, and Ditylenchus. We highlighted that 5 Mg ha−1 of rice husk biochar can be the optimum in shaping the nematode community

A Single-Cell and Feeder-Free Culture System for Monkey Embryonic Stem Cells

<div><p>Primate pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), hold great potential for research and application in regenerative medicine and drug discovery. To maximize primate PSC potential, a practical system is required for generating desired functional cells and reproducible differentiation techniques. Much progress regarding their culture systems has been reported to date; however, better methods would still be required for their practical use, particularly in industrial and clinical fields. Here we report a new single-cell and feeder-free culture system for primate PSCs, the key feature of which is an originally formulated serum-free medium containing FGF and activin. In this culture system, cynomolgus monkey ESCs can be passaged many times by single-cell dissociation with traditional trypsin treatment and can be propagated with a high proliferation rate as a monolayer without any feeder cells; further, typical PSC properties and genomic stability can be retained. In addition, it has been demonstrated that monkey ESCs maintained in the culture system can be used for various experiments such as <i>in vitro</i> differentiation and gene manipulation. Thus, compared with the conventional culture system, monkey ESCs grown in the aforementioned culture system can serve as a cell source with the following practical advantages: simple, stable, and easy cell maintenance; gene manipulation; cryopreservation; and desired differentiation. We propose that this culture system can serve as a reliable platform to prepare primate PSCs useful for future research and application.</p></div

Karyotype analysis.

<p>CMK6_SFF and CMK970 cells retain the normal karyotype, male 40 XY. Red arrows indicate the Y chromosome.</p

Neuronal differentiation.

<p>CMK6_SFF cells were differentiated into cortical neurons. A. Immunocytochemical analysis. Scale bar = 50 µm. B–D. NMDA-induced Ca²⁺ influx. NMDA (with 10 µM glycine) induced a concentration-dependent increase in [Ca²⁺]_i (B). MK-801 (C, NMDA receptor antagonist) and ifenprodil (D, NR2B-specific antagonist) decreased NMDA (10 µM)-induced Ca²⁺ influx.</p