Stepwise fate conversion of supporting cells to sensory hair cells in the chick auditory epithelium

In contrast to mammals, the avian cochlea, specifically the basilar papilla, can regenerate sensory hair cells, which involves fate conversion of supporting cells to hair cells. To determine the mechanisms for converting supporting cells to hair cells, we used single-cell RNA sequencing during hair cell regeneration in explant cultures of chick basilar papillae. We identified dynamic changes in the gene expression of supporting cells, and the pseudotime trajectory analysis demonstrated the stepwise fate conversion from supporting cells to hair cells. Initially, supporting cell identity was erased and transition to the precursor state occurred. A subsequent gain in hair cell identity progressed together with downregulation of precursor-state genes. Transforming growth factor β receptor 1-mediated signaling was involved in induction of the initial step, and its inhibition resulted in suppression of hair cell regeneration. Our data provide new insights for understanding fate conversion from supporting cells to hair cells in avian basilar papillae

Formation of Galactic Center Magnetic Loops

A survey for the molecular clouds in the Galaxy with NANTEN mm telescope has discovered molecular loops in the Galactic center region. The loops show monotonic gradients of the line of sight velocity along the loops and the large velocity dispersions towards their foot points. It is suggested that these loops are explained in terms of the buoyant rise of magnetic loops due to the Parker instability. We have carried out global three-dimensional magneto-hydrodynamic simulations of the gas disk in the Galactic center. The gravitational potential is approximated by the axisymmetric potential proposed by Miyamoto & Nagai (1975). At the initial state, we assume a warm (~ 10^4 K) gas torus threaded by azimuthal magnetic fields. Self-gravity and radiative cooling of the gas are ignored. We found that buoyantly rising magnetic loops are formed above the differentially rotating, magnetically turbulent disk. By analyzing the results of global MHD simulations, we have identified individual loops, about 180 in the upper half of the disk, and studied their statistical properties such as their length, width, height, and velocity distributions along the loops. Typical length and height of a loop are 1kpc and 200pc, respectively. The line of sight velocity changes linearly along a loop and shows large dispersions around the foot-points. Numerical results indicate that loops emerge preferentially from the region where magnetic pressure is large. We argue that these properties are consistent with those of the molecular loops discovered by NANTEN.Comment: 16pages, 10figures. Accepted for publication in PASJ. Replace to higher resolution versio

The added value of non-contrast 3-Tesla MRI for the pre-operative localization of hyperparathyroidism

OBJECTIVE: We investigated the efficacy of non-contrast 3-Tesla MR imaging added to the combination of sestamibi with99mTc (MIBI) scintigraphy and Ultrasonography (US) for the pre-operative localization of Primary Hyperparathyroidism (PHPT) lesions. METHODS: A total of 34 parathyroid glands, including nine normal glands, were examined with MIBI, US, and non-contrast 3-Tesla MRI. MRI was performed with the acquisition of T1- and T2-weighted images and fat-suppressed T2-weighted images. We calculated the sensitivities of MIBI, US, and the 'additional' MRI, with knowledge of the former two modalities' results. RESULTS: For the diagnosis of PHPT lesions, the sensitivity values of MIBI, US, and additional MRI were 88.0% (22/25), 84.0% (21/25), and 92.0% (23/25), respectively. Normal glands were not visualized with any modality (0/9). One lesion was detected neither with US nor MRI, but only with MIBI, with the limitation that MIBI represented no more than laterality. The two glands not identified in MRI were 4 mm and 6 mm in their size, which are within the range of normal gland's size. Two lesions were not detected with US or MIBI but were visualized with the additional MRI, which indicated that the MRI contributed an 8.0% (2/25) improvement of sensitivity, compared from that of US. Fat-suppressed T2-weighted images were useful in the identification of parathyroid lesions, as these images helped to differentiate between the lesion and the adjacent tissue. CONCLUSION: Additional non-contrast 3-Tesla MRI was a useful adjunctive tool for localization of PHPT, which improved the sensitivity of the pre-operative localization of PHPT lesions. Fat-suppressed T2-weighted images contributed to their identification. LEVEL VI: Evidence from a single descriptive or qualitative study

Pharmacokinetic and pharmacodynamic evaluation of a new sustained-release capsules using starch-sponge matrix (SSM) release system for nifedipine in rats

We conducted a performance assessment study for a new sustained-release capsule including starch-sponge matrix (SSM). The SSM, which is a support medium for drug release, was made from 2.5% cornstarch glue by means of freezing dry method. The SSM capsule was applied for nifedipine (NFP), a calcium channel blocker, and evaluated pharmacokinetic and pharmacodynamic (PK/PD) profiles of NFP after intraduodenal administration of SSM capsules including 2.5 or 5.0 mg of NFP per capsule to rats. Plasma NFP concentrations from the SSM capsules showed dose-dependent increases with a Michaelis-Menten like behavior over 360 minutes after intraduodenal administration. The values of area under the concentration vs. time curve from time zero to 360 min (AUC0-360) of NFP declined in making SSM capsules as compared to control capsules due to a simple physical mixture of NFP and cornstarch, but the values of mean residence time (MRT0-360) extended and abidingness of SSM capsules were admitted with dose-dependent manner. As for a PD parameter, the mean arterial blood pressure (mABP) derived from the SSM capsules showed 15~20% decrease of baseline within 120min after intraduodenal administration, and thereafter the mABP in 2.5 mg SSM capsule was gradually recovered, while a relatively smooth and even change was found in the mABP at 5.0 mg SSM capsule. The relationships between plasma NFP concentration and sampling-time corresponding mABP after intraduodenal administration of SSM capsules showed no rapid change in the mABP, indicating that a sustained-release mechanism due to the SSM functions sufficiently to avoid a fluctuating blood pressure accompanied by going up and down of plasma levels of NFP. The SSM capsules exhibited a sustained-release pharmacokinetics of NFP, and made the fluctuation range with blood pressure small compared to the physical mixture preparations. Thus, it was evidenced that the SSM capsule is useful device to provide a sustained-release systems and optimal therapeutic efficacy of drugs.Keywords: Controlled-release, Cornstarch, Matrix, Nifedipine, Pharmacokinetics, Pharmacodynamics

A Lipid Bilayer Formed on a Hydrogel Bead for Single Ion Channel Recordings

Ion channel proteins play important roles in various cell functions, making them attractive drug targets. Artificial lipid bilayer recording is a technique used to measure the ion transport activities of channel proteins with high sensitivity and accuracy. However, the measurement efficiency is low. In order to improve the efficiency, we developed a method that allows us to form bilayers on a hydrogel bead and record channel currents promptly. We tested our system by measuring the activities of various types of channels, including gramicidin, alamethicin, alpha-hemolysin, a voltage-dependent anion channel 1 (VDAC1), a voltage- and calcium-activated large conductance potassium channel (BK channel), and a potassium channel from Streptomyces lividans (KcsA channel). We confirmed the ability for enhanced measurement efficiency and measurement system miniaturizion

Robust folding of a de novo designed ideal protein even with most of the core mutated to valine

Rie Koga, Mami Yamamoto, Takahiro Kosugi, Naohiro Kobayashi, Toshihiko Sugiki, Toshimichi Fujiwara, Nobuyasu Koga, Robust folding of a de novo designed ideal protein even with most of the core mutated to valine. Proceedings of the National Academy of Sciences. 117 (49), 31149-31156 (2020). DOI: 10.1073/pnas.2002120117

Discovery of Molecular Loop 3 in the Galactic Center: Evidence for a Positive-Velocity Magnetically Floated Loop towards L=355∘−359∘L=355^\circ-359^\circ

We have discovered a molecular dome-like feature towards $355^{\circ} \leq l \leq 359^{\circ}$ and $0^{\circ} \leq b \leq 2^{\circ}$. The large velocity dispersions of 50--100 km s$^{-1}$ of this feature are much larger than those in the Galactic disk and indicate that the feature is located in the Galactic center, probably within $\sim1$ kpc of Sgr A$^{*}$. The distribution has a projected length of $\sim600$ pc and height of $\sim300$ pc from the Galactic disk and shows a large-scale monotonic velocity gradient of $\sim130$ km s $^{-1}$ per $\sim600$ pc. The feature is also associated with HI gas having a more continuous spatial and velocity distribution than that of $^{12}$CO. We interpret the feature as a magnetically floated loop similar to loops 1 and 2 and name it "loop 3". Loop 3 is similar to loops 1 and 2 in its height and length but is different from loops 1 and 2 in that the inner part of loop 3 is filled with molecular emission. We have identified two foot points at the both ends of loop 3. HI, $^{12}$CO and $^{13}$CO datasets were used to estimate the total mass and kinetic energy of loop 3 to be \sim3.0 \times 10^{6} \Mo and $\sim1.7 \times 10^{52}$ ergs. The huge size, velocity dispersions and energy are consistent with the magnetic origin the Parker instability as in case of loops 1 and 2 but is difficult to be explained by multiple stellar explosions. We argue that loop 3 is in an earlier evolutionary phase than loops 1 and 2 based on the inner-filled morphology and the relative weakness of the foot points. This discovery indicates that the western part of the nuclear gas disk of $\sim1$ kpc radius is dominated by the three well-developed magnetically floated loops and suggests that the dynamics of the nuclear gas disk is strongly affected by the magnetic instabilities.Comment: 30 pages, 10 figures. High resolution figures are available at http://www.a.phys.nagoya-u.ac.jp/~motosuji/fujishita09_figs

A Detailed Observational Study of Molecular Loops 1 and 2 in the Galactic Center

Fukui et al. (2006) discovered two huge molecular loops in the Galactic center located in (l, b) ~ (355 deg-359 deg, 0 deg-2 deg) in a large velocity range of -180-40 km s^-1. Following the discovery, we present detailed observational properties of the two loops based on NANTEN 12CO(J=1-0) and 13CO(J=1-0) datasets at 10 pc resolution including a complete set of velocity channel distributions and comparisons with HI and dust emissions as well as with the other broad molecular features. We find new features on smaller scales in the loops including helical distributions in the loop tops and vertical spurs. The loops have counterparts of the HI gas indicating that the loops include atomic gas. The IRAS far infrared emission is also associated with the loops and was used to derive an X-factor of 0.7(+/-0.1){\times}10^20 cm^-2 (K km s^-1)^-1 to convert the 12CO intensity into the total molecular hydrogen column density. From the 12CO, 13CO, H I and dust datasets we estimated the total mass of loops 1 and 2 to be ~1.4 {\times} 106 Msun and ~1.9 {\times} 10^6 Msun, respectively, where the H I mass corresponds to ~10-20% of the total mass and the total kinetic energy of the two loops to be ~10^52 ergs. An analysis of the kinematics of the loops yields that the loops are rotating at ~47 km s-1 and expanding at ~141 km s^-1 at a radius of 670 pc from the center. Fukui et al. (2006) presented a model that the loops are created by the magnetic flotation due to the Parker instability with an estimated magnetic field strength of ~150 {\mu}G. We present comparisons with the recent numerical simulations of the magnetized nuclear disk by Machida et al. (2009) and Takahashi et al. (2009) and show that the theoretical results are in good agreements with the observations. The helical distributions also suggest that some magnetic instability plays a role similarly to the solar helical features.Comment: 40 pages, 22 figures, submitted to publication in PAS