Is Autophagy Involved in Pepper Fruit Ripening?

Autophagy is a universal self-degradation process involved in the removal and recycling of cellular constituents and organelles; however, little is known about its possible role in fruit ripening, in which the oxidation of lipids and proteins and changes in the metabolism of different cellular organelles occur. In this work, we analyzed several markers of autophagy in two critical maturation stages of pepper (Capsicum annuum L.) fruits where variations due to ripening become clearly visible. Using two commercial varieties that ripen to yellow and red fruits respectively, we studied changes in the gene expression and protein content of several autophagy (ATG) components, ATG4 activity, as well as the autophagy receptor NBR1 and the proteases LON1 and LON2. Additionally, the presence of intravacuolar vesicles was analyzed by electron microscopy. Altogether, our data reveal that autophagy plays a role in the metabolic changes which occur during ripening in the two studied varieties, suggesting that this process may be critical to acquiring final optimal quality of pepper fruits.

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Nitric oxide-dependent regulation of sweet pepper fruit ripening

Journal of Experimental Botany ◽

10.1093/jxb/erz136 ◽

2019 ◽

Vol 70 (17) ◽

pp. 4557-4570 ◽

Cited By ~ 19

Author(s):

Salvador González-Gordo ◽

Rocío Bautista ◽

M Gonzalo Claros ◽

Amanda Cañas ◽

José M Palma ◽

...

Keyword(s):

Nitric Oxide ◽

Fruit Ripening ◽

Biochemical Analysis ◽

Sweet Pepper ◽

Supporting Evidence ◽

Functional Categories ◽

Fatty Acid Profiling ◽

Pepper Fruit

Abstract Ripening is a complex physiological process that involves changes in reactive nitrogen and oxygen species that govern the shelf-life and quality of fruits. Nitric oxide (NO)-dependent changes in the sweet pepper fruit transcriptome were determined by treating fruits at the initial breaking point stage with NO gas. Fruits were also harvested at the immature (green) and ripe (red) stages. Fruit ripening in the absence of NO resulted in changes in the abundance of 8805 transcripts whose function could be identified. Among these, functional clusters associated with reactive oxygen/nitrogen species and lipid metabolism were significantly modified. NO treatment resulted in the differential expression of 498 genes framed within these functional categories. Biochemical analysis revealed that NO treatment resulted in changes in fatty acid profiling, glutathione and proline contents, and the extent of lipid peroxidation, as well as increases in the activity of ascorbate peroxidase and lipoxygenase. These data provide supporting evidence for the crucial role of NO in the ripening of pepper fruit.

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The Current Situation in Chemical Stabilization of Biological Materials

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100093912 ◽

1972 ◽

Vol 30 ◽

pp. 132-133

Author(s):

John H. Luft

Keyword(s):

Electron Microscopy ◽

Electron Microscope ◽

Osmium Tetroxide ◽

Molecular Level ◽

Cross Linking ◽

Chemical Stabilization ◽

Specimen Preparation ◽

Sufficient Condition ◽

Radio Telescopes

With information processing devices such as radio telescopes, microscopes or hi-fi systems, the quality of the output often is limited by distortion or noise introduced at the input stage of the device. This analogy can be extended usefully to specimen preparation for the electron microscope; fixation, which initiates the processing sequence, is the single most important step and, unfortunately, is the least well understood. Although there is an abundance of fixation mixtures recommended in the light microscopy literature, osmium tetroxide and glutaraldehyde are favored for electron microscopy. These fixatives react vigorously with proteins at the molecular level. There is clear evidence for the cross-linking of proteins both by osmium tetroxide and glutaraldehyde and cross-linking may be a necessary if not sufficient condition to define fixatives as a class.

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Characterization of homoepitaxial diamond films by Electron microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100088713 ◽

1991 ◽

Vol 49 ◽

pp. 880-881

Author(s):

D.P. Malta ◽

S.A. Willard ◽

R.A. Rudder ◽

G.C. Hudson ◽

J.B. Posthill ◽

...

Keyword(s):

Electron Microscopy ◽

Surface Defects ◽

Substrate Surface ◽

Diamond Films ◽

Chemical Vapor ◽

Polycrystalline Diamond ◽

Large Degree ◽

Rf Plasma ◽

Semiconducting Diamond

Semiconducting diamond films have the potential for use as a material in which to build active electronic devices capable of operating at high temperatures or in high radiation environments. A major goal of current device-related diamond research is to achieve a high quality epitaxial film on an inexpensive, readily available, non-native substrate. One step in the process of achieving this goal is understanding the nucleation and growth processes of diamond films on diamond substrates. Electron microscopy has already proven invaluable for assessing polycrystalline diamond films grown on nonnative surfaces.The quality of the grown diamond film depends on several factors, one of which is the quality of the diamond substrate. Substrates commercially available today have often been found to have scratched surfaces resulting from the polishing process (Fig. 1a). Electron beam-induced current (EBIC) imaging shows that electrically active sub-surface defects can be present to a large degree (Fig. 1c). Growth of homoepitaxial diamond films by rf plasma-enhanced chemical vapor deposition (PECVD) has been found to planarize the scratched substrate surface (Fig. 1b).

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The annealed (0001) α-alumina surface and its influence on thin-film growth

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100138051 ◽

1995 ◽

Vol 53 ◽

pp. 336-337

Author(s):

Michael W. Bench ◽

Paul G. Kotula ◽

C. Barry Carter

Keyword(s):

Electron Microscopy ◽

Surface Energy ◽

Film Growth ◽

Thin Film Growth ◽

Energy Calculations ◽

Transmission Electron ◽

Reflection Electron Microscopy ◽

Low Energy Electron ◽

Surface Nature

The growth of semiconductors, superconductors, metals, and other insulators has been investigated using alumina substrates in a variety of orientations. The surface state of the alumina (for example surface reconstruction and step nature) can be expected to affect the growth nature and quality of the epilayers. As such, the surface nature has been studied using a number of techniques including low energy electron diffraction (LEED), reflection electron microscopy (REM), transmission electron microscopy (TEM), molecular dynamics computer simulations, and also by theoretical surface energy calculations. In the (0001) orientation, the bulk alumina lattice can be thought of as a layered structure with A1-A1-O stacking. This gives three possible terminations of the bulk alumina lattice, with theoretical surface energy calculations suggesting that termination should occur between the Al layers. Thus, the lattice often has been described as being made up of layers of (Al-O-Al) unit stacking sequences. There is a 180° rotation in the surface symmetry of successive layers and a total of six layers are required to form the alumina unit cell.

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Scanning Secondary Electron Microscopy of Myofibrils Using Transmission Techniques

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100116401 ◽

1975 ◽

Vol 33 ◽

pp. 556-557

Author(s):

M. K. Lamvik

Keyword(s):

Electron Microscopy ◽

Electron Microscope ◽

Carbon Films ◽

Photographic Recording ◽

Transmission Electron ◽

Thin Carbon ◽

The Common ◽

Scanning Electron ◽

Better Than ◽

Cellular Organelles

When observing small objects such as cellular organelles by scanning electron microscopy, it is often valuable to use the techniques of transmission electron microscopy. The common practice of mounting and coating for SEM may not always be necessary. These possibilities are illustrated using vertebrate skeletal muscle myofibrils.Micrographs for this study were made using a Hitachi HFS-2 scanning electron microscope, with photographic recording usually done at 60 seconds per frame. The instrument was operated at 25 kV, with a specimen chamber vacuum usually better than 10-7 torr. Myofibrils were obtained from rabbit back muscle using the method of Zak et al. To show the component filaments of this contractile organelle, the myofibrils were partially disrupted by agitation in a relaxing medium. A brief centrifugation was done to clear the solution of most of the undisrupted myofibrils before a drop was placed on the grid. Standard 3 mm transmission electron microscope grids covered with thin carbon films were used in this study.

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Stain contamination and embedding in electron microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100141986 ◽

1986 ◽

Vol 44 ◽

pp. 50-53

Author(s):

Hilton H. Mollenhauer

Keyword(s):

Electron Microscopy ◽

Information Retrieval ◽

Epoxy Resins ◽

Image Contrast ◽

Sample Preservation ◽

Factors Associated ◽

Amount Of Information ◽

Electron Microscopical ◽

Contrast Information

Many factors (e.g., resolution of microscope, type of tissue, and preparation of sample) affect electron microscopical images and alter the amount of information that can be retrieved from a specimen. Of interest in this report are those factors associated with the evaluation of epoxy embedded tissues. In this context, informational retrieval is dependant, in part, on the ability to “see” sample detail (e.g., contrast) and, in part, on tue quality of sample preservation. Two aspects of this problem will be discussed: 1) epoxy resins and their effect on image contrast, information retrieval, and sample preservation; and 2) the interaction between some stains commonly used for enhancing contrast and information retrieval.

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Methods to Monitor and Improve the Performance of Specimen Holders for Transmission Electron Cryomicroscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100164738 ◽

1996 ◽

Vol 54 ◽

pp. 454-455

Author(s):

B. L. Armbruster ◽

B. Kraus ◽

M. Pan

Keyword(s):

Electron Microscopy ◽

Electron Beam ◽

Electron Beam Irradiation ◽

Beam Irradiation ◽

Quality Of Data ◽

Electron Cryomicroscopy ◽

Thermal Equilibration ◽

Transmission Electron ◽

Electron Microscopes

One goal in electron microscopy of biological specimens is to improve the quality of data to equal the resolution capabilities of modem transmission electron microscopes. Radiation damage and beam- induced movement caused by charging of the sample, low image contrast at high resolution, and sensitivity to external vibration and drift in side entry specimen holders limit the effective resolution one can achieve. Several methods have been developed to address these limitations: cryomethods are widely employed to preserve and stabilize specimens against some of the adverse effects of the vacuum and electron beam irradiation, spot-scan imaging reduces charging and associated beam-induced movement, and energy-filtered imaging removes the “fog” caused by inelastic scattering of electrons which is particularly pronounced in thick specimens.Although most cryoholders can easily achieve a 3.4Å resolution specification, information perpendicular to the goniometer axis may be degraded due to vibration. Absolute drift after mechanical and thermal equilibration as well as drift after movement of a holder may cause loss of resolution in any direction.

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Introduction to Games User Research

10.1093/oso/9780198794844.003.0001 ◽

2018 ◽

Cited By ~ 1

Author(s):

Anders Drachen ◽

Pejman Mirza-Babaei ◽

Lennart E. Nacke

Keyword(s):

User Experience ◽

Video Game ◽

Digital Games ◽

Comprehensive Overview ◽

User Research ◽

Interdisciplinary Field ◽

Present Book ◽

Optimal Quality ◽

Experienced User

This chapter provides an introduction to the field of Games User Research (GUR) and to the present book. GUR is an interdisciplinary field of practice and research concerned with ensuring the optimal quality of usability and user experience in digital games. GUR inevitably involves any aspect of a video game that players interface with, directly or indirectly. This book aims to provide the foundational, accessible, go-to resource for people interested in GUR. It is a community-driven effort—it is written by passionate professionals and researchers in the GUR community as a handbook and guide for everyone interested in user research and games. We aim to provide the most comprehensive overview from an applied perspective, for a person new to GUR, but which is also useful for experienced user researchers.

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Clay-Based Brick Porosity Estimation Using Image Processing Techniques

Journal of Intelligent Systems ◽

10.1515/jisys-2018-0191 ◽

2019 ◽

Vol 29 (1) ◽

pp. 1226-1234

Author(s):

Safa Jida ◽

Hassan Ouallal ◽

Brahim Aksasse ◽

Mohammed Ouanan ◽

Mohamed El Amraoui ◽

...

Keyword(s):

Electron Microscopy ◽

Image Processing ◽

Scanning Electron Microscopy ◽

Computer Vision ◽

Image Processing Techniques ◽

Processing Techniques ◽

Porosity Estimation ◽

Microscopy Images ◽

Scanning Electron

Abstract This work intends to apprehend and emphasize the contribution of image-processing techniques and computer vision in the treatment of clay-based material known in Meknes region. One of the various characteristics used to describe clay in a qualitative manner is porosity, as it is considered one of the properties that with “kill or cure” effectiveness. For this purpose, we use scanning electron microscopy images, as they are considered the most powerful tool for characterising the quality of the microscopic pore structure of porous materials. We present various existing methods of segmentation, as we are interested only in pore regions. The results show good matching between physical estimation and Voronoi diagram-based porosity estimation.

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Cryo-EM model validation recommendations based on outcomes of the 2019 EMDataResource challenge

Nature Methods ◽

10.1038/s41592-020-01051-w ◽

2021 ◽

Vol 18 (2) ◽

pp. 156-164 ◽

Cited By ~ 4

Author(s):

Catherine L. Lawson ◽

Andriy Kryshtafovych ◽

Paul D. Adams ◽

Pavel V. Afonine ◽

Matthew L. Baker ◽

...

Keyword(s):

Electron Microscopy ◽

Structure Data ◽

Model Evaluation ◽

Data Bank ◽

High Accuracy ◽

Atomic Resolution ◽

Software Developers ◽

Data Archives ◽

Modeling Software

AbstractThis paper describes outcomes of the 2019 Cryo-EM Model Challenge. The goals were to (1) assess the quality of models that can be produced from cryogenic electron microscopy (cryo-EM) maps using current modeling software, (2) evaluate reproducibility of modeling results from different software developers and users and (3) compare performance of current metrics used for model evaluation, particularly Fit-to-Map metrics, with focus on near-atomic resolution. Our findings demonstrate the relatively high accuracy and reproducibility of cryo-EM models derived by 13 participating teams from four benchmark maps, including three forming a resolution series (1.8 to 3.1 Å). The results permit specific recommendations to be made about validating near-atomic cryo-EM structures both in the context of individual experiments and structure data archives such as the Protein Data Bank. We recommend the adoption of multiple scoring parameters to provide full and objective annotation and assessment of the model, reflective of the observed cryo-EM map density.

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