Radial alignment of microtubules through tubulin polymerization in an evaporating droplet

© The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Keya, J. J., Kudoh, H., Kabir, A. M. R., Inoue, D., Miyamoto, N., Tani, T., Kakugo, A., & Shikinaka, K. Radial alignment of microtubules through tubulin polymerization in an evaporating droplet. Plos One, 15(4), (2020): e0231352, doi:10.1371/journal.pone.0231352.We report the formation of spherulites from droplets of highly concentrated tubulin solution via nucleation and subsequent polymerization to microtubules (MTs) under water evaporation by heating. Radial alignment of MTs in the spherulites was confirmed by the optical properties of the spherulites observed using polarized optical microscopy and fluorescence microscopy. Temperature and concentration of tubulins were found as important parameters to control the spherulite pattern formation of MTs where evaporation plays a significant role. The alignment of MTs was regulated reversibly by temperature induced polymerization and depolymerization of tubulins. The formation of the MTs patterns was also confirmed at the molecular level from the small angle X-ray measurements. This work provides a simple method for obtaining radially aligned arrays of MTs.Fund receiver: Akira Kakugo Grant-in-Aid for Scientific Research on Innovative Areas (Grant Nos. JP24104004 and 18H05423) and a Grant-in-Aid for Scientific Research (A) (Grant No. 18H03673) from kaken. NO - The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscrip

Scanning SQUID microscope system for geological samples: system integration and initial evaluation

We have developed a high-resolution scanning superconducting quantum interference device (SQUID) microscope for imaging the magnetic field of geological samples at room temperature. In this paper, we provide details about the scanning SQUID microscope system, including the magnetically shielded box (MSB), the XYZ stage, data acquisition by the system, and initial evaluation of the system. The background noise in a two-layered PC permalloy MSB is approximately 40–50 pT. The long-term drift of the system is approximately ≥1 nT, which can be reduced by drift correction for each measurement line. The stroke of the XYZ stage is 100 mm × 100 mm with an accuracy of ~10 µm, which was confirmed by laser interferometry. A SQUID chip has a pick-up area of 200 µm × 200 µm with an inner hole of 30 µm × 30 µm. The sensitivity is 722.6 nT/V. The flux-locked loop has four gains, i.e., ×1, ×10, ×100, and ×500. An analog-to-digital converter allows analog voltage input in the range of about ±7.5 V in 0.6-mV steps. The maximum dynamic range is approximately ±5400 nT, and the minimum digitizable magnetic field is ~0.9 pT. The sensor-to-sample distance is measured with a precision line current, which gives the minimum of ~200 µm. Considering the size of pick-up coil, sensor-to-sample distance, and the accuracy of XYZ stage, spacial resolution of the system is ~200 µm. We developed the software used to measure the sensor-to-sample distance with line scan data, and the software to acquire data and control the XYZ stage for scanning. We also demonstrate the registration of the magnetic image relative to the optical image by using a pair of point sources placed on the corners of a sample holder outside of a thin section placed in the middle of the sample holder. Considering the minimum noise estimate of the current system, the theoretical detection limit of a single magnetic dipole is ~1 × 10-14 Am2. The new instrument is a powerful tool that could be used in various applications in paleomagnetism such as ultrafine-scale magnetostratigraphy and single-crystal paleomagnetism

Mesoscopic Architectures Made of Electrically Charged Binary Colloidal Nanosheets in Aqueous System

Inorganic layered materials can be converted to colloidal liquid crystals through exfoliation into inorganic nanosheets, and binary nanosheet colloids exhibit rich phase behavior characterized by multiphase coexistence. In particular, niobate–clay binary nanosheet colloids are characterized by phase separation at a mesoscopic (∼several tens of micrometers) scale whereas they are apparently homogeneous at a macroscopic scale. Although the mesoscopic structure of the niobate–clay binary colloid is advantageous to realize unusual photochemical functions, the structure itself has not been clearly demonstrated in real space. The present study investigated the structure of niobate–clay binary nanosheet colloids in detail. Four clay nanosheets (hectorite, saponite, fluorohectorite, and tetrasilisic mica) with different lateral sizes were compared. Small-angle X-ray scattering (SAXS) indicated lamellar ordering of niobate nanosheets in the binary colloid. The basal spacing of the lamellar phase was reduced by increasing the concentration of clay nanosheets, indicating the compression of the liquid crystalline niobate phase by the isotropic clay phase. Scattering and fluorescence microscope observations using confocal laser scanning microscopy (CLSM) demonstrated the phase separation of niobate and clay nanosheets in real space. Niobate nanosheets assembled into domains of several tens of micrometers whereas clay nanosheets were located in voids between the niobate domains. The results clearly confirmed the spatial separation of two nanosheets and the phase separation at a mesoscopic scale. Distribution of clay nanosheets is dependent on the employed clay nanosheets; the nanosheets with large lateral length are more localized or assembled. This is in harmony with larger basal spacings of niobate lamellar phase for large clay particles. Although three-dimensional compression of the niobate phase by the coexisting clay phase was observed at low clay concentrations, the basal spacing of niobate phase was almost constant irrespective of niobate concentrations at high clay concentrations, which was ascribed to competition of compression by clay phase and restoring of the niobate phase

Polycystic Kidney Disease in the Medaka (Oryzias latipes) pc Mutant Caused by a Mutation in the Gli-Similar3 (glis3) Gene

Polycystic kidney disease (PKD) is a common hereditary disease in humans. Recent studies have shown an increasing number of ciliary genes that are involved in the pathogenesis of PKD. In this study, the Gli-similar3 (glis3) gene was identified as the causal gene of the medaka pc mutant, a model of PKD. In the pc mutant, a transposon was found to be inserted into the fourth intron of the pc/glis3 gene, causing aberrant splicing of the pc/glis3 mRNA and thus a putatively truncated protein with a defective zinc finger domain. pc/glis3 mRNA is expressed in the epithelial cells of the renal tubules and ducts of the pronephros and mesonephros, and also in the pancreas. Antisense oligonucleotide-mediated knockdown of pc/glis3 resulted in cyst formation in the pronephric tubules of medaka fry. Although three other glis family members, glis1a, glis1b and glis2, were found in the medaka genome, none were expressed in the embryonic or larval kidney. In the pc mutant, the urine flow rate in the pronephros was significantly reduced, which was considered to be a direct cause of renal cyst formation. The cilia on the surface of the renal tubular epithelium were significantly shorter in the pc mutant than in wild-type, suggesting that shortened cilia resulted in a decrease in driving force and, in turn, a reduction in urine flow rate. Most importantly, EGFP-tagged pc/glis3 protein localized in primary cilia as well as in the nucleus when expressed in mouse renal epithelial cells, indicating a strong connection between pc/glis3 and ciliary function. Unlike human patients with GLIS3 mutations, the medaka pc mutant shows none of the symptoms of a pancreatic phenotype, such as impaired insulin expression and/or diabetes, suggesting that the pc mutant may be suitable for use as a kidney-specific model for human GLIS3 patients

リンショウケンキュウ ト エキガクケンキュウ ニ オケル ロンブン ノ シツ ヲ タカメルタメ ノ コクサイドウコウ ヒト ヲ タイショウ ト シタ ケンキュウデザイン ノ エビデンス・グレーディング

本研究は,人を対象とした治療や健康増進の効果を明らかにする研究のエビデンスを示す方法をレビューすることを目的とした。疫学や臨床研究においては,研究デザインによって,エビデンスのグレーディング(grading : 格付け)が行われている。各種の研究デザインにおける論文を評価したり,論文の質を高めるためにチェックリストや声明が開発されている。また,人を対象とした介入研究を実施する場合には,パブリケーション・バイアスの減少,研究の倫理,臨床試験参加の推進につながるため,事前の臨床試験登録(clinical trial registry : CTR)が必要である。世界的な11の医学雑誌からなる「国際医学雑誌編集者委員会(International Committee of Medical Journal Editors : ICMJE)」は,臨床試験の登録(clinical trial registry : CTR)を論文投稿の条件としている。疫学研究や臨床研究を実施する場合には,こうした動向を理解する必要がある。This narrative reviews the means of effectively evaluating evidence in the outcome of clinical trial and quality of health care. The grading of evidence in clinical and epidemiologic studies has been provided to improve the study design and to raise the quality of publications. Check lists and guidelines are developed to reduce publication bias, and clinical trial registry has the potential to facilitate ethics and to increase participation of clinical trials. The International Committee of Medical Journal Editors has proposed that only clinical trials are published that are registered at inception. These trends are to be appreciated when epidemiologic or clinical study is conducted

無機層状結晶を利用したカチオン性シアニン色素のナノおよびマクロスケールでの組織化

制度:新 ; 文部省報告番号:乙1858号 ; 学位の種類:博士(工学) ; 授与年月日:2004-02-5 ; 早大学位記番号:新3713早稲田大

Liquid Crystalline Behavior and Related Properties of Colloidal Systems of Inorganic Oxide Nanosheets

Inorganic layered crystals exemplified by clay minerals can be exfoliated in solvents to form colloidal dispersions of extremely thin inorganic layers that are called nanosheets. The obtained “nanosheet colloids” form lyotropic liquid crystals because of the highly anisotropic shape of the nanosheets. This system is a rare example of liquid crystals consisting of inorganic crystalline mesogens. Nanosheet colloids of photocatalytically active semiconducting oxides can exhibit unusual photoresponses that are not observed for organic liquid crystals. This review summarizes experimental work on the phase behavior of the nanosheet colloids as well as photochemical reactions observed in the clay and semiconducting nanosheets system