scholarly journals Correlative Light and Electron Microscopy (CLEM) Analysis of Nuclear Reorganization Induced by Clustered DNA Damage Upon Charged Particle Irradiation

2020 ◽  
Vol 21 (6) ◽  
pp. 1911 ◽  
Author(s):  
Susanne Tonnemacher ◽  
Mikhail Eltsov ◽  
Burkhard Jakob
Keyword(s):  
Electron Microscopy ◽  
Dna Damage ◽  
Light And Electron Microscopy ◽  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Dna Double Strand Breaks ◽  
Nuclear Reorganization ◽  
Particle Irradiation ◽  
Transmission Electron ◽  
Radiation Induced

Chromatin architecture plays major roles in gene regulation as well as in the repair of DNA damaged by endogenous or exogenous factors, such as after radiation. Opening up the chromatin might provide the necessary accessibility for the recruitment and binding of repair factors, thus facilitating timely and correct repair. The observed formation of ionizing radiation-induced foci (IRIF) of factors, such as 53BP1, upon induction of DNA double-strand breaks have been recently linked to local chromatin decompaction. Using correlative light and electron microscopy (CLEM) in combination with DNA-specific contrasting for transmission electron microscopy or tomography, we are able to show that at the ultrastructural level, these DNA damage domains reveal a chromatin compaction and organization not distinguishable from regular euchromatin upon irradiation with carbon or iron ions. Low Density Areas (LDAs) at sites of particle-induced DNA damage, as observed after unspecific uranyl acetate (UA)-staining, are thus unlikely to represent pure chromatin decompaction. RNA-specific terbium-citrate (Tb) staining suggests rather a reduced RNA density contributing to the LDA phenotype. Our observations are discussed in the view of liquid-like phase separation as one of the mechanisms of regulating DNA repair.

Effects of O3 on submucosal gland of bonnet monkey trachea

1983 ◽  
Vol 41 ◽  
pp. 678-679
Author(s):  
S. Yamashiro ◽  
D. Wilson ◽  
J. St. George ◽  
D. Hyde ◽  
C. Plopper ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Periodic Acid ◽  
Light And Electron Microscopy ◽  
Lung Parenchyma ◽  
Uranyl Acetate ◽  
Upper Airways ◽  
Periodic Acid Schiff ◽  
Transmission Electron ◽  
Bonnet Monkey ◽  
Submucosal Glands

In the past, ozone inhalation studies have focused on the lower airways and lung parenchyma. The purpose of this study was to evaluate the effects of ozone on submucosal glands of upper airways. Six adult male bonnet monkeys were exposed to 0.64 ppm ozone continuously for 7 days, and three were exposed to chamber conditions without ozone. The animals were exsanguinated under barbiturate anesthesia. The trachea and lung were fixed by airway infusion of Karnovsky's fixative, which was adjusted to pH 7.4 and 440 milliOsmols. Sagittal sections of ventral trachea were embedded in glycol methacrylate and Araldite 502 for light and electron microscopy. One micrometer methacrylate sections were stained with Alcian blue-periodic acid Schiff (AB/PAS). Selected areas of Araldite-embedded tissue were sectioned for transmission electron microscopy, stained with uranyl acetate and lead citrate and examined with a Zeiss EM 10. Volume percentages of the lumen, granular and nongranular regions of fhe gland and the duct wall, respectively, were estimated by stereologic methods on AB/PAS stained sections.

Annealing Of Radiation Damage in Tungsten

1970 ◽  
Vol 28 ◽  
pp. 412-413
Author(s):  
Robert C. Rau ◽  
John Moteff
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Radiation Damage ◽  
Thermal Annealing ◽  
Neutron Fluence ◽  
Dislocation Loops ◽  
Defect Clusters ◽  
Polycrystalline Tungsten ◽  
Transmission Electron ◽  
Radiation Induced

Transmission electron microscopy has been used to study the thermal annealing of radiation induced defect clusters in polycrystalline tungsten. Specimens were taken from cylindrical tensile bars which had been irradiated to a fast (E > 1 MeV) neutron fluence of 4.2 × 1019 n/cm2 at 70°C, annealed for one hour at various temperatures in argon, and tensile tested at 240°C in helium. Foils from both the unstressed button heads and the reduced areas near the fracture were examined.Figure 1 shows typical microstructures in button head foils. In the unannealed condition, Fig. 1(a), a dispersion of fine dot clusters was present. Annealing at 435°C, Fig. 1(b), produced an apparent slight decrease in cluster concentration, but annealing at 740°C, Fig. 1(C), resulted in a noticeable densification of the clusters. Finally, annealing at 900°C and 1040°C, Figs. 1(d) and (e), caused a definite decrease in cluster concentration and led to the formation of resolvable dislocation loops.

The Ultrastructure of Lymphocytic Hypophysitis

1981 ◽  
Vol 39 ◽  
pp. 466-467
Author(s):  
S.L. Asa ◽  
K. Kovacs ◽  
J. M. Bilbao ◽  
R. G. Josse ◽  
K. Kreines
Keyword(s):  
Electron Microscopy ◽  
Plasma Cells ◽  
Secretory Granules ◽  
Sella Turcica ◽  
Uranyl Acetate ◽  
Lymphocytic Hypophysitis ◽  
Pituitary Cells ◽  
Transmission Electron ◽  
Ultrathin Sections ◽  
Mass Lesions

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).

Investigation of Tickborne Pathogens within Naturally Infected Brown Dog Tick (Ixodidae: Rhipicephalus Sanguineus) in Egypt by Light and Electron Microscopy

2020 ◽  
Keyword(s):  
Electron Microscopy ◽  
Salivary Gland ◽  
Light And Electron Microscopy ◽  
Rhipicephalus Sanguineus ◽  
Mammalian Host ◽  
Babesia Canis ◽  
Brown Dog Tick ◽  
Transmission Electron ◽  
Dog Tick ◽  
Theileria Spp

Tick borne pathogens present a significant health challenge to animals and human because a single tick may transmit multiple pathogens to a mammalian host during feeding. The present study detected tick-borne pathogens from pet dogs. A total of 666 ticks were collected from 144 pet and sheltered dogs in Egypt from April to September 2018. For hemolymph, midgut and salivary gland smears 546 ticks were used as well as 360 egg smears from 120 female tick were examined by light microscope. The infected ticks were prepared for transmission electron microscopy (TEM). Ticks were identified; Rhipicephalus sanguineus. Light microscopy showed infection rates of 44.69%, 68.50% & 15.75%, in hemolymph, midgut and salivary gland, respectively. H. canis recorded the highest rates in hemolymph and midgut (35.89% & 49.82%, respectively), but Theileria spp. was the lowest (0.73% & 2.93%, respectively). In salivary gland smears, Babesia canis. was detected in 13.55% and Theileria spp. in 1.83%. Mixed infection in same tick was recorded in 4.76% &0.37% in midgut and salivary gland smears, respectively. Babesia canis stages were recovered from 15% of egg smears. R. sanguineus was natural infected by Babesia, Theileria, Hepatozoon and Anaplasma phagocytophilum as well as mixed infections of protozoa accompanied by a complicated sign of diseases and failure in accurate diagnosis.

scholarly journals Focused Ion Microbeam Irradiation Induces Clustering of DNA Double-Strand Breaks in Heterochromatin Visualized by Nanoscale-Resolution Electron Microscopy

2021 ◽  
Vol 22 (14) ◽  
pp. 7638
Author(s):  
Yvonne Lorat ◽  
Judith Reindl ◽  
Anna Isermann ◽  
Christian Rübe ◽  
Anna A. Friedl ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Dna Damage ◽  
Spatial Clustering ◽  
Dna Lesions ◽  
Double Strand Breaks ◽  
Stimulated Emission ◽  
Nuclear Chromatin ◽  
Carbon Ions ◽  
Dna Double Strand Breaks ◽  
Strand Breaks

Background: Charged-particle radiotherapy is an emerging treatment modality for radioresistant tumors. The enhanced effectiveness of high-energy particles (such as heavy ions) has been related to the spatial clustering of DNA lesions due to highly localized energy deposition. Here, DNA damage patterns induced by single and multiple carbon ions were analyzed in the nuclear chromatin environment by different high-resolution microscopy approaches. Material and Methods: Using the heavy-ion microbeam SNAKE, fibroblast monolayers were irradiated with defined numbers of carbon ions (1/10/100 ions per pulse, ipp) focused to micrometer-sized stripes or spots. Radiation-induced lesions were visualized as DNA damage foci (γH2AX, 53BP1) by conventional fluorescence and stimulated emission depletion (STED) microscopy. At micro- and nanoscale level, DNA double-strand breaks (DSBs) were visualized within their chromatin context by labeling the Ku heterodimer. Single and clustered pKu70-labeled DSBs were quantified in euchromatic and heterochromatic regions at 0.1 h, 5 h and 24 h post-IR by transmission electron microscopy (TEM). Results: Increasing numbers of carbon ions per beam spot enhanced spatial clustering of DNA lesions and increased damage complexity with two or more DSBs in close proximity. This effect was detectable in euchromatin, but was much more pronounced in heterochromatin. Analyzing the dynamics of damage processing, our findings indicate that euchromatic DSBs were processed efficiently and repaired in a timely manner. In heterochromatin, by contrast, the number of clustered DSBs continuously increased further over the first hours following IR exposure, indicating the challenging task for the cell to process highly clustered DSBs appropriately. Conclusion: Increasing numbers of carbon ions applied to sub-nuclear chromatin regions enhanced the spatial clustering of DSBs and increased damage complexity, this being more pronounced in heterochromatic regions. Inefficient processing of clustered DSBs may explain the enhanced therapeutic efficacy of particle-based radiotherapy in cancer treatment.

Radiation Induced Formation of Acrylated Palm Oil (APO) Nanoparticles Using Cetyltrimethylammonium Bromide Microemulsion System

2011 ◽  
Vol 364 ◽  
pp. 278-282
Author(s):  
Rida Tajau ◽  
Dahlan Khairul Mohd. Zaman ◽  
Mohd Hilmi Mahmood ◽  
Wan Md Zin Wan Yunus ◽  
Hashim Kamaruddin
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Palm Oil ◽  
Chemical Structure ◽  
Cetyltrimethylammonium Bromide ◽  
Control Process ◽  
Microemulsion System ◽  
Transmission Electron ◽  
Radiation Induced ◽  
The Fourier Transform

In this study, we report the preparation of Acrylated Palm Oil (APO) nanoparticles using aqueous Cetyltrimethylammonium bromide (CTAB) microemulsion system. This microemulsion system which contains the dispersed APO nanodroplets was subjected to the gamma irradiation to induce the formation of the crosslinked APO nanoparticles. The dynamic light scattering (DLS), the Fourier Transform Infrared (FTIR) spectrocospy and the Transmission Electron Microscopy (TEM) were used to characterize the size and the chemical structure of the nanoparticle. Sized of the APO nanoor microparticle can be varied depended on the volumes of the modified palm oil and the irradiation doses. Their size is in the range of 73 to 100 nanometer (nm) after irradiation using gamma irradiator. This radiation-induced method provides a free initiator induced and easy to control process as compared to the classical or chemical initiator process.

scholarly journals Localization of ‘Candidatus Liberibacter solanacearum’ and Evidence for Surface Appendages in the Potato Psyllid Vector

2016 ◽  
Vol 106 (2) ◽  
pp. 142-154 ◽  
Author(s):  
J. M. Cicero ◽  
T. W. Fisher ◽  
J. K. Brown
Keyword(s):  
Electron Microscopy ◽  
Light And Electron Microscopy ◽  
Fluorescence Labeling ◽  
Potato Psyllid ◽  
Bactericera Cockerelli ◽  
Visual Identification ◽  
Transmission Electron ◽  
Candidatus Liberibacter ◽  
Identification Of Bacteria

The potato psyllid Bactericera cockerelli is implicated as the vector of the causal agent of zebra chip of potato and vein-greening of tomato diseases. Until now, visual identification of bacteria in the genus ‘Candidatus Liberibacter’ has relied on direct imaging by light and electron microscopy without labeling, or with whole-organ fluorescence labeling only. In this study, aldehyde fixative followed by a coagulant fixative, was used to process adult psyllids for transmission electron microscopy (TEM) colloidal gold in situ hybridization experiments. Results indicated that ‘Ca. Liberibacter solanacearum’ (CLso)-specific DNA probes annealed to a bacterium that formed extensive, monocultural biofilms on gut, salivary gland, and oral region tissues, confirming that it is one morphotype of potentially others, that is rod-shaped, approximately 2.5 µm in diameter and of variable length, and has a rough, granular cytosol. In addition, CLso, prepared from shredded midguts, and negatively stained for TEM, possessed pili- and flagella-like surface appendages. Genes implicating coding capacity for both types of surface structures are encoded in the CLso genome sequence. Neither type was seen for CLso associated with biofilms within or on digestive organs, suggesting that their production is stimulated only in certain environments, putatively, in the gut during adhesion leading to multiplication, and in hemolymph to afford systemic invasion.

scholarly journals Tetrahymena Meiotic Nuclear Reorganization Is Induced by a Checkpoint Kinase–dependent Response to DNA Damage

2009 ◽  
Vol 20 (9) ◽  
pp. 2428-2437 ◽  
Author(s):  
Josef Loidl ◽  
Kazufumi Mochizuki
Keyword(s):  
Dna Damage ◽  
Kinase Inhibitors ◽  
Signal Transduction Pathway ◽  
Dna Lesions ◽  
Meiotic Pairing ◽  
Dna Double Strand Breaks ◽  
Strand Breaks ◽  
Nuclear Reorganization ◽  
Bivalent Formation ◽  
Atr Kinase

In the ciliate Tetrahymena, meiotic micronuclei (MICs) undergo extreme elongation, and meiotic pairing and recombination take place within these elongated nuclei (the “crescents”). We have previously shown that elongation does not occur in the absence of Spo11p-induced DNA double-strand breaks (DSBs). Here we show that elongation is restored in spo11Δ mutants by various DNA-damaging agents including ones that may not cause DSBs to a notable extent. MIC elongation following Spo11p-induced DSBs or artificially induced DNA lesions is probably a DNA-damage response mediated by a phosphokinase signal transduction pathway, since it is suppressed by the ATM/ATR kinase inhibitors caffeine and wortmannin and by knocking out Tetrahymena's ATR orthologue. MIC elongation occurs concomitantly with the movement of centromeres away from the telomeric pole of the MIC. This DNA damage–dependent reorganization of the MIC helps to arrange homologous chromosomes alongside each other but is not sufficient for exact pairing. Thus, Spo11p contributes to bivalent formation in two ways: by creating a favorable spatial disposition of homologues and by stabilizing pairing by crossovers. The polarized chromosome orientation inside the crescent resembles the conserved meiotic bouquet, and crescent and bouquet also share the putative function of aiding meiotic pairing. However, they are regulated differently because in Tetrahymena, DSBs are required for entering rather than exiting this stage.

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