Structural analysis of photosynthetic membranes by cryo-electron tomography of intact Rhodopseudomonas viridis cells

During the photosynthetic process, highly organized membranal assemblies convert light into biochemical energy with high efficiency. We have used whole-mount cryo-electron tomography to study the intracellular architecture of the photosynthetic membranes of the anaerobic purple photosynthetic bacterium Rhodopseudomonas viridis, as well as the organization of the photosynthetic units within the membranes. Three-dimensional reconstruction demonstrates a continuity of the plasma membrane with the photosynthetic membranes that form tunnel-like structures with an average diameter of 31 nm ± 8 nm at the connection sites. The spacing between the photosynthetic membranes at their cytoplasmic faces was found to be 11 nm, thus enforcing a highly close packaging of the photosynthetic membranes. Analysis of successive tomographic slices allowed for derivation of the spacing between adjacent photosynthetic core complexes from a single-layered photosynthetic membrane, in situ. This analysis suggests that most, if not all, photosynthetic membranes in R. viridis are characterized by a similar two-dimensional hexagonal lattice organization.     

Expedited approaches to whole cell electron tomography and organelle mark-up in situ in high-pressure frozen pancreatic islets

We have developed a simplified, efficient approach for the 3D reconstruction and analysis of mammalian cells in toto by electron microscope tomography (ET), to provide quantitative information regarding ‘global’ cellular organization at 15–20 nm resolution. Two insulin-secreting beta cells—deemed ‘functionally equivalent’ by virtue of their location at the periphery of the same pancreatic islet—were reconstructed in their entirety in 3D after fast-freezing/freeze-substitution/plastic embedment in situ within a glucose-stimulated islet of Langerhans isolated intact from mouse pancreata. These cellular reconstructions have afforded several unique insights into fundamental structure–function relationships among key organelles involved in the biosynthesis and release of the crucial metabolic hormone, insulin, that could not be provided by other methods. The Golgi ribbon, mitochondria and insulin secretory granules in each cell were segmented for comparative analysis. We propose that relative differences between the two cells in terms of the number, dimensions and spatial distribution (and for mitochondria, also the extent of branching) of these organelles per cubic micron of cellular volume reflects differences in the two cells’ individual capacity (and/or readiness) to respond to secretagogue stimulation, reflected by an apparent inverse relationship between the number/size of insulin secretory granules versus the number/size of mitochondria and the Golgi ribbon. We discuss the advantages of this approach for quantitative cellular ET of mammalian cells, briefly discuss its application relevant to other complementary techniques, and summarize future strategies for overcoming some of its current limitations.     

Optical Doppler tomography based on a field programmable gate array

We report the design of and results obtained by using a field programmable gate array (FPGA) to digitally process optical Doppler tomography signals. The processor fits into the analog signal path in an existing optical coherence tomography setup. We demonstrate both Doppler frequency and envelope extraction using the Hilbert transform, all in a single FPGA. An FPGA implementation has certain advantages over general purpose digital signal processor (DSP) due to the fact that the processing elements operate in parallel as opposed to the DSP, which is primarily a sequential processor.     

Coronary artery anomalies detected by MSCT-coronary angiography in the adult

Background. Before coronary evaluation by modern imaging techniques was feasible, premorbid diagnoses of coronary artery anomalies (CAAs) were usually made fortuitously by invasive coronary angiography (ICA). However, this technique is limited by its invasive and projectional nature. Coronary magnetic resonance angiography (CMRA) and multi-slice computed tomography (MSCT) broadened clinical information by enabling visualisation of the coronary arteries in their anatomical environment. Methods. This case series visualises and reviews anomalous coronary artery from the opposite sinus (ACAOS) and coronary artery fistulae. All CAAs were detected by means of 64-slice dual source computed tomography after 1000 cardiac scans at the Erasmus MC, Rotterdam, the Netherlands. Results. Eight ACAOS cases, one anomalous left coronary artery from the pulmonary artery (ALCAPA) and one congenital aneurysm of an aortic sinus were found. Seven out often detected CAAs were considered malignant whereas three CAAs of the ACAOS type (retroaortic path) were considered benign. Significant coronary artery disease was found in three out of eight ACAOS cases. In one of the ACAOS cases complete evaluation of the anomalous coronary artery was limited by motion artifacts. All five cases of right ACAOS were referred for MSCT because the right coronary artery could not be located by invasive angiography. Conclusion. All CAAs were easy to diagnose because of 3D imaging and high temporal and spatial resolution. High resolution made it possible to not only depict coronary artery abnormalities, but also to quantify luminal and vessel properties such as stenosis grade, aspects of plaque, anomalous vessel length, luminal area ratio and the asymmetry ratio. Because of its comprehensiveness, MSCT can be an effective imaging modality in patients suspected of coronary artery abnormalities caused by coronary artery disease, CAAs, or a combination of both. (Neth Heart J 2008;16:369-75.)     

中国数字化可视人体鞍上池的横断面解剖与CT、MRI对照研究

目的:探讨鞍上池在中国数字化可视人体(Chinese visible human,CVH)与CT、MRI上的横断面解剖形态学表现。方法:选择做64层螺旋CT和MRI头部检查的健康志愿者各60例,获得5mm层厚横断面图像。从第2例中国数字化可视人体数据集中选取与CT、MRI相对应层面的头部薄层连续横断面标本图像,对照观察鞍上池在CVH、MRI与CT图像上的正常解剖形态、毗邻及内部结构。结果:CVH图像上,鞍上池表现为六角形和五角形两种形状。CVH薄层横断面图像能连续、清晰地显示鞍上池的正常形态、毗邻及内部结构。60例CT及MRI图像上,鞍上池全部显示,但解剖结构均不及CVH清晰。鞍上池在CT、MRI横断面图像上形状变化更大,以六角形最多,五角形次之,四角形最少,相应毗邻及内部结构也有所不同。六角形鞍上池在CVH、CT、MRI上有良好的对应关系,五角形鞍上池部分相匹配,CVH图像上无四角形鞍上池。结论:通过与CT、MRI进行对照研究,中国数字化可视人体能为颅脑疾病的影像识别和诊断提供断层解剖学依据。     

Echocardiographic Abnormalities in Normotensive Obese Patients: Relationship with Visceral Fat

Objective: To evaluate the relationship of echocardiographic characteristics and visceral adipose tissue (VAT) distribution in normotensive obese patients. Research Methods and Procedures: Echocardiographic parameters were assessed in 28 normotensive obese patients [7 men, 21 women, mean age, 43.2 years; mean body mass index (BMI), 37.2 kg/m²; 10 with impaired glucose tolerance (IGT); 6 with type 2 diabetes] and 18 sex‐ and age‐matched healthy, normal‐weight controls (4 men, 14 women; mean age, 45.8 years; mean BMI, 22.4 kg/m²) by an M‐mode, color‐doppler videofluoroscope. VAT in the obese patients was assessed by computed tomography (at L4 level). Results: The obese patients had a significantly larger internal diastolic left ventricular (LV) diameter (p < 0.05), a thicker end‐diastolic septum (p < 0.001) and posterior wall (p < 0.001), a greater indexed (g/m^2.7) LV mass (p < 0.001), a higher atrial diastolic filling wave velocity (p < 0.001), a lower ratio between early and atrial diastolic filling wave velocities (p < 0.01), and a prolonged isovolumic relaxation time (p < 0.05). End‐diastolic septum and posterior wall thickness and the LV mass were significantly greater in patients with a VAT area >130 cm² than with <130 cm². In the multivariate regression analysis, only VAT (p < 0.0001), waist‐to‐hip ratio (p < 0.001), and sex (p < 0.001) were associated with the most important echocardiographic alterations. Discussion: The morphological and functional echocardiographic alterations usually found in normotensive obese patients closely correlate with the amount of intra‐abdominal fat deposition, even in the presence of diabetes or IGT.     

Study of the Morphological and Functional State of Higher Plant Tissues by Optical Coherence Microscopy and Optical Coherence Tomography

Comparative analysis of two optical methods—optical coherence tomography (OCT) and optical coherence microscopy (OCM)—was made for vital visualization of plant tissues in tomato (Lycopersicon esculentum Mill), spiderwort (Tradescantia pallida (Rose) D. Hunt), orach (Atriplex sp.), and leaves and seeds of medium starwort (Stellaria media L.). The obtained OCT- and OCM-images allowed the morphological and functional state of plant tissues to be assessed in vivo. A higher spatial resolution of the OCM method, as compared to OCT method, allowed plant morphological structures to be identified with greater confidence. The morphological and functional state of tissues can be monitored with a time resolution of 1–4 s in intact plants, without removing them from the habitat.__________Translated from Fiziologiya Rastenii, Vol. 52, No. 4, 2005, pp. 628–634.Original Russian Text Copyright © 2005 by Kutis, Sapozhnikova, Kuranov, Kamenskii.     

Anatomy of the Cranial Endocast of the Bottlenose Dolphin, Tursiops truncatus, Based on HRXCT

Endocranial surfaces, volumes, and interconnectivities of extant and fossil odontocetes potentially offer information on the general architecture of the brain and on the structure of the specialized cetacean circulatory system. Although conventional methods for acquiring such data have generally involved invasive preparation of the specimen, particularly in the case of fossils, new tomographic technologies afford nondestructive access to these internal morphologies. In this study we used high-resolution X-ray computed tomography (HRXCT) to scan a skull of the extant Tursiops truncatus (Cetacea: Odontoceti). We processed the data to reveal the cranial endocast and details of internal skeletal architecture (data at www.digimorph.org). Major features that can be discerned include aspects of the specimen's hypertrophied retia mirabilia, the major canals and openings of the cranial cavity, and the relationship of the brain and endocranial circulatory structures to the surrounding skeleton. CT data also provide information on the shape of the brain that may be lost in conventional anatomical preparations, and readily provide volumetric and linear measurements of the endocast and its individual segments. These results demonstrate the utility of HRXCT for interpreting the internal cranial anatomy of both extant and fossil cetaceans.     

强直性脊柱炎骶髂关节CT MRI表现

目的 :旨在评估强直性脊柱炎 (AS)患者骶髂关节炎的MR影像学特征。并比较X线平片、CT和MR影像在诊断骶髂关节炎中的作用。方法 :搜集 1 2例AS患者 ,分别行X线平片、CT和MR检查。增强前MR扫描序列包括SET1 W1 、FSET2 WI和梯度回波的准T2 (GRT2 WI)。增强后MR扫描序列参数与增强前SET2 WI相同。另选 9例志愿者 ,行MR平扫检查。结果 :8例志愿者 1 6个骶髂关节的T1 WI和T2 WI可直接显示正常骶髂关节软骨表现为线形或点样的中等信号影。 9例志愿者 1 1个骶髂关节的骨髓内可见局灶性脂肪沉积。 1 2例AS患者 2 1个骶髂关节可见软骨异常 ,表现为T1 WI和T2 WI上正常线形中等信号的软骨影像消失代之以不规则增粗或扭曲样中等信号。患者组CT表现为Ⅱ～Ⅳ级骶髂关节炎 ,即关节软骨下骨板硬化 ,侵蚀囊变 ,关节间隙宽窄不均或部分强直 ,韧带部受累。分析表明 :在骶髂关节炎的诊断方面 ,MR和CT明显优于X线平片 (P <0 .0 0 1 )。结论 :同X线片比较虽CT和MRI均有助于AS骶髂关节炎的诊断 ,但MR影像可显示CT和X线所不能显示的软骨异常和骨髓内水肿改变 ;骨髓内脂肪沉积可属正常变异 ,扭曲样中等强度信号应视为软骨的异常征象。     

Electron microscopy of high pressure frozen samples: bridging the gap between cellular ultrastructure and atomic resolution

Transmission electron microscopy has provided most of what is known about the ultrastructural organization of tissues, cells, and organelles. Due to tremendous advances in crystallography and magnetic resonance imaging, almost any protein can now be modeled at atomic resolution. To fully understand the workings of biological “nanomachines” it is necessary to obtain images of intact macromolecular assemblies in situ. Although the resolution power of electron microscopes is on the atomic scale, in biological samples artifacts introduced by aldehyde fixation, dehydration and staining, but also section thickness reduces it to some nanometers. Cryofixation by high pressure freezing circumvents many of the artifacts since it allows vitrifying biological samples of about 200 μm in thickness and immobilizes complex macromolecular assemblies in their native state in situ. To exploit the perfect structural preservation of frozen hydrated sections, sophisticated instruments are needed, e.g., high voltage electron microscopes equipped with precise goniometers that work at low temperature and digital cameras of high sensitivity and pixel number. With them, it is possible to generate high resolution tomograms, i.e., 3D views of subcellular structures. This review describes theory and applications of the high pressure cryofixation methodology and compares its results with those of conventional procedures. Moreover, recent findings will be discussed showing that molecular models of proteins can be fitted into depicted organellar ultrastructure of images of frozen hydrated sections. High pressure freezing of tissue is the base which may lead to precise models of macromolecular assemblies in situ, and thus to a better understanding of the function of complex cellular structures.