In situ visualization of a simple bipartite kinetochore with a single microtubule attachment in Giardia intestinalis (Metamonada)

To understand general features in evolution of kinetochore organization, investigating a wide range of mitotic mechanisms in various non-model eukaryotes is necessary. A binucleate flagellate Giardia intestinalis is a representative of highly divergent eukaryotic lineage of Metamonads. FIB/SEM tomography was used to investigate ultrastructural details of its mitotic architecture, including kinetochores. Giardia undergoes semi-open mitosis, with the nuclear envelope remaining intact except for polar fenestrae, allowing microtubules to enter the nucleoplasm. At the onset of mitosis, the nuclear envelope bends inward, forming a concave depression at the spindle poles. Spindle microtubules emanate from a cytoplasmic fuzzy microtubule organizing center near the flagellar basal bodies. Kinetochoral microtubules enter the nucleoplasm and bind to kinetochores. A small bipartite kinetochore composed of a dense inner disk, approximately 46 nm in diameter, and a two-armed outer fork, is attached to just one microtubule. To our knowledge, this is the first in situ evidence of a one-microtubule attachment to a kinetochore, which could represent a basic eukaryotic situation.     

Multiscale Morphological and Electrical Characterization of Charge Transport Limitations to the Power Performance of Positive Electrode Blends for Lithium‐Ion Batteries

In this work, exhaustive characterizations of 3D geometries of LiNi_1/3Mn_1/3Co_1/3O₂ (NMC), LiFePO₄ (LFP), and NMC/LFP blended electrodes are undertaken for rational interpretation of their measured electrical properties and electrochemical performance. X‐ray tomography and focused ion beam in combination with scanning electron microscopy tomography are used for a multiscale analysis of electrodes 3D geometries. Their multiscale electrical properties are measured by using broadband dielectric spectroscopy. Finally, discharge rate performance are measured and analyzed by simple, yet efficient methods. It allows us to discriminate between electronic and ionic wirings as the performance limiting factors, depending on the discharge rate. This approach is a unique exhaustive analysis of the experimental relationships between the electrochemical behavior, the transport properties within the electrode, and its 3D geometry.     

奥扎格雷钠联合川芎嗪注射液对急性脑梗死患者血清同型半胱氨酸、C 反应蛋白及纤维蛋白原水平的影响

目的:探讨奥扎格雷钠联合川芎嗪注射液对急性脑梗死患者血清同型半胱氨酸(HCY)、C反应蛋白(CRP)及纤维蛋白原(FIB)的水平的影响。方法:选择我院收治的急性脑梗死患者68例,随机分为实验组与对照组,每组34例。对照组患者给予奥扎格雷钠治疗,实验组患者给予奥扎格雷钠联合使用川芎嗪注射液治疗,观察并比较治疗前后两组患者血清HCY,CRP及FIB的水平以及颈内动脉粥样狭窄程度的变化和临床总有效率。结果:与治疗前相比,两组患者血清HCY,CRP及FIB水平均显著降低,颈内动脉粥样硬化情况均好转(P0.05);且与对照组相比,实验组患者血清HCY,CRP及FIB水平较低,颈内动脉粥样硬化情况较轻(P0.05);与对照组相比,实验组患者临床总有效率较高(P0.05)。结论:奥扎格雷钠联合川芎嗪注射液治疗可显著提高急性脑梗死积的临床疗效,并能够降低患者颈内动脉粥样狭窄程度,其机制可能与降低血清HCY,CRP及纤维蛋白原水平有关。     

替米沙坦联合叶酸对老年H型高血压合并2型糖尿病患者凝血功能及肾功的影响

目的:观察替米沙坦联合叶酸治疗对老年H型高血压合并2型糖尿病患者凝血功能及肾功的影响.为老年H型高血压合并2型糖尿病患者的临床治疗以及延缓病程提供理论依据.方法:选择66例符合标准的老年H型高血压合并2型糖尿病患者,测定血压,胰岛素敏感指数,糖化血红蛋白,同型半胱氨酸,凝血指标及尿白蛋白排泄率(UAE)等基线数据.上述患者随机分为对照组及实验组,分别接受替米沙坦80 mg/d及替米沙坦80 mg/d+叶酸片0.8 mg/d治疗后,于24周后复查上述指标,并与用药前进行对比研究.结果:对照组及实验组患者均较治疗前血压下降,糖化血红蛋白、纤维蛋白原、尿白蛋白排泄率明显下降(P＜0.05),胰岛素敏感指数较治疗前升高(P＜0.05).经治疗后,实验组患者较对照组血压、糖化血红蛋白、同型半胱氨酸、纤维蛋白原、尿白蛋白排泄率下降均低于对照组(P＜0.05),胰岛素敏感指数较对照组升高(P＜0.05).结论:对于老年H型高血压合并2型糖尿病患者替米沙坦联合叶酸治疗较单纯替米沙坦治疗具有更良好的降压作用,增加胰岛素敏感性,改善高凝状态,改善肾功.     

Who's In and Who's Out—Compositional Control of Biomolecular Condensates

Biomolecular condensates are two- and three-dimensional compartments in eukaryotic cells that concentrate specific collections of molecules without an encapsulating membrane. Many condensates behave as dynamic liquids and appear to form through liquid–liquid phase separation driven by weak, multivalent interactions between macromolecules. In this review, we discuss current models and data regarding the control of condensate composition, and we describe our current understanding of the composition of representative condensates including PML nuclear bodies, P-bodies, stress granules, the nucleolus, and two-dimensional membrane localized LAT and nephrin clusters. Specific interactions, such as interactions between modular binding domains, weaker interactions between intrinsically disorder regions and nucleic acid base pairing, and nonspecific interactions, such as electrostatic interactions and hydrophobic interactions, influence condensate composition. Understanding how specific condensate composition is determined is essential to understanding condensates as biochemical entities and ultimately discerning their cellular and organismic functions.     

Methods for Physical Characterization of Phase-Separated Bodies and Membrane-less Organelles

Membrane-less organelles are cellular structures which arise through the phenomenon of phase separation. This process enables compartmentalization of specific sets of macromolecules (e.g., proteins, nucleic acids), thereby regulating cellular processes by increasing local concentration, and modulating the structure and dynamics of their constituents. Understanding the connection between structure, material properties and function of membrane-less organelles requires inter-disciplinary approaches, which address length and timescales that span several orders of magnitude (e.g., Ångstroms to micrometer, picoseconds to hours). In this review, we discuss the wide variety of methods that have been applied to characterize the morphology, rheology, structure and dynamics of membrane-less organelles and their components, in vitro and in live cells.     

Towards native-state imaging in biological context in the electron microscope

Modern cell biology is reliant on light and fluorescence microscopy for analysis of cells, tissues and protein localisation. However, these powerful techniques are ultimately limited in resolution by the wavelength of light. Electron microscopes offer much greater resolution due to the shorter effective wavelength of electrons, allowing direct imaging of sub-cellular architecture. The harsh environment of the electron microscope chamber and the properties of the electron beam have led to complex chemical and mechanical preparation techniques, which distance biological samples from their native state and complicate data interpretation. Here we describe recent advances in sample preparation and instrumentation, which push the boundaries of high-resolution imaging. Cryopreparation, cryoelectron microscopy and environmental scanning electron microscopy strive to image samples in near native state. Advances in correlative microscopy and markers enable high-resolution localisation of proteins. Innovation in microscope design has pushed the boundaries of resolution to atomic scale, whilst automatic acquisition of high-resolution electron microscopy data through large volumes is finally able to place ultrastructure in biological context.     

Ligand Incorporation into Protein Microcrystals for MicroED by On-Grid Soaking

1. Download : Download high-res image (237KB)
2. Download : Download full-size image

     

The multi‐scale architecture of mammalian sperm flagella and implications for ciliary motility

Motile cilia are molecular machines used by a myriad of eukaryotic cells to swim through fluid environments. However, available molecular structures represent only a handful of cell types, limiting our understanding of how cilia are modified to support motility in diverse media. Here, we use cryo‐focused ion beam milling‐enabled cryo‐electron tomography to image sperm flagella from three mammalian species. We resolve in‐cell structures of centrioles, axonemal doublets, central pair apparatus, and endpiece singlets, revealing novel protofilament‐bridging microtubule inner proteins throughout the flagellum. We present native structures of the flagellar base, which is crucial for shaping the flagellar beat. We show that outer dense fibers are directly coupled to microtubule doublets in the principal piece but not in the midpiece. Thus, mammalian sperm flagella are ornamented across scales, from protofilament‐bracing structures reinforcing microtubules at the nano‐scale to accessory structures that impose micron‐scale asymmetries on the entire assembly. Our structures provide vital foundations for linking molecular structure to ciliary motility and evolution.     

Crystallographic characterization of the crossed lamellar structure in the bivalve Meretrix lamarckii using electron beam techniques

To understand the formation mechanism of crossed lamellar structures in molluskan shells, the crystallographic structural features in the shell of a bivalve, Meretrix lamarckii, were investigated using scanning electron microscopy, electron backscattered diffraction, and transmission electron microscopy with a focused ion beam sample preparation technique. Approximately 0.5 μm-thick lamellae (the second-order units) are piled up obliquely toward the growth direction to form the first-order unit and the obliquity is inverted between adjacent units along the shell thickness direction. The first-order units originate around the center of the shell, initially growing parallel to the shell and subsequently curving toward the inner or outer surfaces. The lamellae consist of aragonite granular and columnar layers, which group together to adopt the same crystal orientation forming crystallographic units (crystallites). Multiple {1 1 0} twins are common both in the granular and columnar layers. The crystallite c-axis is parallel to the columns and is inclined at angles 0–50° from the lamellar normal (dispersing among individual lamellae), toward the shell growth direction. Probably, the directions of the a- and b-axes are random in the lamellae, showing no specific orientation.