scholarly journals Identification of putative viroplasms within banana cells infected by banana streak MY virus

2020 ◽  
Vol 101 (12) ◽  
pp. 1305-1312
Author(s):  
Thu Ha Ngo ◽  
Richard Webb ◽  
Kathleen S. Crew ◽  
Megan E. Vance ◽  
John E. Thomas ◽  
...  

The badnavirus replication cycle is poorly understood and most knowledge is based on extrapolations from model viruses such as Cauliflower mosaic virus (CaMV). However, in contrast to CaMV, badnaviruses are thought not to produce viroplasms and therefore it has been a mystery as to where virion assembly occurs. In this study, ultrathin sections of a banana leaf infected with a badnavirus, banana streak MY virus (BSMYV), were examined by transmission electron microscopy. Electron-dense inclusion bodies (EDIBs) were sporadically distributed in parenchymatous tissues of the leaf, most commonly in the palisade and spongy mesophyll cells. These EDIBs had a characteristic structure, comprising an electron-dense core, a single, encircling lacuna and an outer ring of electron-dense material. However, much less frequently, EDIBs with two or three lacunae were observed. In the outer ring, densely packed virions were visible with a shape and size consistent with that expected for badnaviruses. Immunogold labelling was done with primary antibodies that detected the N-terminus of the capsid protein and strong labelling of the outer ring but not the central core or lacuna was observed. It is concluded that the EDIBs that were observed are equivalent in function to the viroplasms of CaMV, although obviously different in composition as there is not a paralogue of the transactivation/viroplasm protein in the badnavirus genome. It is postulated that production of a viroplasm could be a conserved characteristic of all members of the Caulimoviridae.

Author(s):  
S.L. Asa ◽  
K. Kovacs ◽  
J. M. Bilbao ◽  
R. G. Josse ◽  
K. Kreines

Seven cases of lymphocytic hypophysitis in women have been reported previously in association with various degrees of hypopituitarism. We report two pregnant patients who presented with mass lesions of the sella turcica, clinically mimicking pituitary adenoma. However, pathologic examination revealed extensive infiltration of the anterior pituitary by lymphocytes and plasma cells with destruction of the gland. To our knowledge, the ultrastructural features of lymphocytic hypophysitis have not been studied so far.For transmission electron microscopy, tissue from surgical specimens was fixed in glutaraldehyde, postfixed in OsO4, dehydrated and embedded in epoxy-resin. Ultrathin sections were stained with uranyl acetate and lead citrate and examined with a Philips 300 electron microscope.Electron microscopy revealed adenohypophysial cells of all types exhibiting varying degrees of injury. In the areas of most dense inflammatory cell infiltration pituitary cells contained large lysosomal bodies fusing with secretory granules (Fig. 1), as well as increased numbers of swollen mitochondria, indicating oncocytic transformation (Fig. 2).


Author(s):  
Kenichi Takaya

Mast cell and basophil granules of the vertebrate contain heparin or related sulfated proteoglycans. Histamine is also present in mammalian mast cells and basophils. However, no histamine is detected in mast cell granules of the amphibian or fish, while it is shown in those of reptiles and birds A quantitative x-ray microanalysis of mast cell granules of fresh frozen dried ultrathin sections of the tongue of Wistar rats and tree frogs disclosed high concentrations of sulfur in rat mast cell granules and those of sulfur and magnesium in the tree frog granules. Their concentrations in tree frog mast cell granules were closely correlated (r=0.94).Fresh frozen dried ultrathin sections and fresh air-dried prints of the tree frog tongue and spleen and young red-eared turtle (ca. 6 g) spleen and heart blood were examined by a quantitative energy-dispersive x-ray microanalysis (X-650, Kevex-7000) for the element constituents of the granules of mast cells and basophils. The specimens were observed by transmission electron microscopy (TEM) (80-200 kV) and followed by scanning transmission electron microscopy (STEM) under an analytical electron microscope (X-650) at an acceleration voltage of 40 kV and a specimen current of 0.2 nA. A spot analysis was performed in a STEM mode for 100 s at a specimen current of 2 nA on the mast cell and basophil granules and other areas of the cells. Histamine was examined by the o-phthalaldehyde method.


Author(s):  
K. S. Zaychuk ◽  
M. H. Chen ◽  
C. Hiruki

Wheat spot mosaic (WSpM), which frequently occurs with wheat streak mosaic virus was first reported in 1956 from Alberta. Singly isolated, WSpM causes chlorotic spots, chlorosis, stunting, and sometimes death of the wheat plants. The vector responsible for transmission is the eriophyid mite, Eriophyes tulipae Kiefer. The examination of leaf ultrastructure by electron microscopy has revealed double membrane bound bodies (DMBB’s) 0.1-0.2 μm in diameter. Dispersed fibrils within these bodies suggested the presence of nucleic acid. However, neither ribosomes characteristic of bacteria, mycoplasma and the psittacosis group of organisms nor an electron dense core characteristic of many viruses was commonly evident.In an attempt to determine if the DMBB’s contain nucleic acids, RNase A, DNase I, and lactoferrin protein were conjugated with 10 nm colloidal gold as previously described. Young root and leaf tissues from WSpM-affected wheat plants were fixed in glutaraldehyde, postfixed in osmium tetroxide,and embedded in Spurr’s resin.


Author(s):  
Ellen Holm Nielsen

In secretory cells a dense and complex network of actin filaments is seen in the subplasmalemmal space attached to the cell membrane. During exocytosis this network is undergoing a rearrangement facilitating access of granules to plasma membrane in order that fusion of the membranes can take place. A filamentous network related to secretory granules has been reported, but its structural organization and composition have not been examined, although this network may be important for exocytosis.Samples of peritoneal mast cells were frozen at -70°C and thawed at 4°C in order to rupture the cells in such a gentle way that the granule membrane is still intact. Unruptured and ruptured cells were fixed in 2% paraformaldehyde and 0.075% glutaraldehyde, dehydrated in ethanol. For TEM (transmission electron microscopy) cells were embedded in Lowicryl K4M at -35°C and for SEM (scanning electron microscopy) they were placed on copper blocks, critical point dried and coated. For immunoelectron microscopy ultrathin sections were incubated with monoclonal anti-actin and colloidal gold labelled IgM. Ruptured cells were also placed on cover glasses, prefixed, and incubated with anti-actin and colloidal gold labelled IgM.


Author(s):  
Blayne Fritz ◽  
Stanley J. Naides ◽  
Kenneth Moore

The pseudoreplica method of staining viral particles for visualization by transmission electron microscopy is a very popular technique. The ability to concentrate clinical specimens while semi-embedding viral particles makes it especially well suited for morphologic and diagnostic virology. Immunolabelling viral particles with colloidal gold is a technique frequently employed by both research and diagnostic virologists. We have characterized a procedure which provides the advantage of both by modifying and combining pseudoreplica staining and immunogold labelling.Modification of specimen retrieval and delay of staining allows us to utilize pseudoreplica processed specimens within our standard immunogold labelling protocol. In brief, we absorbed samples onto 2% agarose, added.25% Formvar and wicked dry. We then floated the Formvar-virus film onto double distilled water, added grids and retrieved with parafilm. The Formvar-virus specimens were then treated as thin tissue sections within our standard two stage immunolabelling protocol. Following completion of immunogold labelling; each grid was negatively stained with phosphotungstic acid or uranyl acetate contrast stains.


1993 ◽  
Vol 71 (11) ◽  
pp. 2282-2290 ◽  
Author(s):  
M. T. Pardue ◽  
J. G. Sivak ◽  
K. M. Kovacs

The corneal anatomy of fin whales (Balaenoptera physalus), minke whales (Balaenoptera acutorostrata), harp seals (Phoca groenlandica), ringed seals (Phoca hispida), and bearded seals (Eriganthus barbatus) was examined to determine if marine mammals have evolved specialized corneas for life in a marine habitat. One to seven eyes of each species were analyzed: paraffin sections stained with haematoxylin and eosin for light microscopy; and ultrathin sections for transmission electron microscopy. All corneas contain the five typical mammalian layers: epithelium, Bowman's layer, stroma, Descemet's membrane, and endothelium. The corneas of these marine mammals are thicker than human corneas because of a thicker stromal layer. The other layers are thinner than those found in humans, except for the epithelial layer in the bearded seal and the cetaceans where it may provide extra protection for the eye during feeding behaviour. The epithelial cells in all corneas studied have an abundance of tonofilaments, which may strengthen the cells and distribute force across the corneal surface. No special organization of collagen fibrils was found in the stroma that would offer protection from ultraviolet radiation or glare for pinnipeds when on ice. The thickness of the sclera in the cetaceans may serve to hold the inner globe of the eye in an elliptical shape, while the thinning of the sclera in the equatorial region in pinnipeds may flatten the eye in air to reduce aerial myopia.


2001 ◽  
Vol 44 (4) ◽  
pp. 405-410 ◽  
Author(s):  
Maria das Graças Sajo ◽  
Silvia Rodrigues Machado

The leaf ultrastructure of five Xyris species were examined using scanning electron microscope (SEM), transmission electron microscope (TEM) and histochemical methods. All studied leaves show some features in epidermis and mesophyll, which were of considerable adaptative significance to drought stress. Such features included the occurrence of a pectic layer on the stomatal guard cells and the presence of a network of pectic compounds in the cuticle. Pectic compunds were also in abundance in lamellated walls of the mesophyll cells and on the inner surface of the sclerified cell walls of the vascular bundle sheaths. There were also specialized chlorenchymatous "peg cells" in the mesophyll and drops of phenolic compounds inside the epidermal cells.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Jaroslav Ondruš ◽  
Pavel Kulich ◽  
Oldřich Sychra ◽  
Pavel Široký

Abstract Neoehrlichia mikurensis is an emerging tick-borne intracellular pathogen causing neoehrlichiosis. Its putative morphology was described in mammalian, but not in tick cells. In this study, we aim to show the presumptive morphology of N. mikurensis in salivary glands of engorged females of Ixodes ricinus. To accomplish this, we collected I. ricinus ticks in a locality with a high N. mikurensis prevalence, allowed them to feed in the artificial in vitro feeding system, dissected salivary glands and screened them by PCR for N. mikurensis and related bacteria. Ultrathin sections of salivary glands positive for N. mikurensis but negative for other pathogens were prepared and examined by transmission electron microscopy. We observed two individual organisms strongly resembling N. mikurensis in mammalian cells as described previously. Both bacteria were of ovoid shape between 0.5–0.8 μm surrounded by the inner cytoplasmic and the rippled outer membrane separated by an irregular electron-lucent periplasmic space. Detection of N. mikurensis in salivary glands of I. ricinus suggests that this bacterium uses the “salivary pathway of transmission” to infect mammals.


2016 ◽  
Vol 682 ◽  
pp. 3-9 ◽  
Author(s):  
Anna Urbańczyk-Gucwa ◽  
Kinga Rodak ◽  
Adam Płachta ◽  
Joanna Sobota ◽  
Zbigniew Rdzawski

The results of the microstructure and hardness investigations of the Cu-0.8Cr alloy after application of severe plastic deformation (SPD) implemented by rolling with the cyclic movement of rolls (RCMR) are presented in this paper. Performed substructure investigations showed that using the RCMR method can refine the microstructure of Cu-0.8Cr alloy to the ultrafine scale. The structure of the Cu-0.8Cr alloy was analyzed using light microscope (LM) and scanning transmission electron microscope (STEM). The quantitative studies of the substructure was performed with "MET-ILO" software, on the basis of images acquired on STEM microscope.


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