Toxicity, Physiological, and Ultrastructural Effects of Arsenic and Cadmium on the Extremophilic Microalga Chlamydomonas acidophila

The cytotoxicity of cadmium (Cd), arsenate (As(V)), and arsenite (As(III)) on a strain of Chlamydomonas acidophila, isolated from the Rio Tinto, an acidic environment containing high metal(l)oid concentrations, was analyzed. We used a broad array of methods to produce complementary information: cell viability and reactive oxygen species (ROS) generation measures, ultrastructural observations, transmission electron microscopy energy dispersive x-ray microanalysis (TEM–XEDS), and gene expression. This acidophilic microorganism was affected differently by the tested metal/metalloid: It showed high resistance to arsenic while Cd was the most toxic heavy metal, showing an LC50 = 1.94 µM. Arsenite was almost four-fold more toxic (LC50= 10.91 mM) than arsenate (LC50 = 41.63 mM). Assessment of ROS generation indicated that both arsenic oxidation states generate superoxide anions. Ultrastructural analysis of exposed cells revealed that stigma, chloroplast, nucleus, and mitochondria were the main toxicity targets. Intense vacuolization and accumulation of energy reserves (starch deposits and lipid droplets) were observed after treatments. Electron-dense intracellular nanoparticle-like formation appeared in two cellular locations: inside cytoplasmic vacuoles and entrapped into the capsule, around each cell. The chemical nature (Cd or As) of these intracellular deposits was confirmed by TEM–XEDS. Additionally, they also contained an unexpected high content in phosphorous, which might support an essential role of poly-phosphates in metal resistance.

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The Protective Effect of Autophagy on DNA Damage in Mouse Spermatocyte-Derived Cells Exposed to 1800 MHz Radiofrequency Electromagnetic Fields

Cellular Physiology and Biochemistry ◽

10.1159/000491660 ◽

2018 ◽

Vol 48 (1) ◽

pp. 29-41 ◽

Cited By ~ 4

Author(s):

Renyan Li ◽

Mingfu Ma ◽

Lianbing Li ◽

Letian Zhao ◽

Tianfeng Zhang ◽

...

Keyword(s):

Dna Damage ◽

Western Blot ◽

Signaling Pathway ◽

Electromagnetic Fields ◽

Ros Generation ◽

Molecular Signaling ◽

Radiofrequency Electromagnetic Fields ◽

Transmission Electron ◽

Amp Activated Protein Kinase

Background/Aims: The effects of exposure to radiofrequency electromagnetic fields (RF-EMFs) on the male reproductive system have raised public concern and studies have shown that exposure to RF-EMFs can induce DNA damage and autophagy. However, there are no related reports on the role of autophagy in DNA damage in spermatocytes, especially after exposure to RF-EMFs. The aim of the present study was to determine the mechanism and role of autophagy induced by RF-EMFs in spermatozoa cells. Methods: Mouse spermatocyte-derived cells (GC-2) were exposed to RF-EMFs 4 W/kg for 24 h. The level of reactive oxygen species (ROS) was determined by ROS assay kit. Comet assay was utilized to detect DNA damage. Autophagy was detected by three indicators: LC3II/LC3I, autophagic vacuoles, and GFP-LC3 dots, which were measured by western blot, transmission electron microscopy, and transfection with GFP-LC3, respectively. The expression of the molecular signaling pathway AMP-activated protein kinase (AMPK)/mTOR was determined by western blot. Results: The results showed that RF-EMFs induced autophagy and DNA damage in GC-2 cells via ROS generation, and the autophagy signaling pathway AMPK/mTOR was activated by ROS generation. Furthermore, following inhibition of autophagy by knockdown of AMPKα, increased DNA damage was observed in GC-2 cells following RF-EMFs exposure, and overexpression of AMPKα promoted autophagy and attenuated DNA damage. Conclusions: These findings demonstrated that the autophagy which was induced by RF-EMFs via the AMPK/mTOR signaling pathway could prevent DNA damage in spermatozoa cells.

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On Future Directions in Pathology

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100053991 ◽

1982 ◽

Vol 40 ◽

pp. 274-277

Author(s):

Benjamin F. Trump ◽

Irene K. Berezesky ◽

Raymond T. Jones

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Clinical Pathology ◽

Future Directions ◽

Diagnostic Pathology ◽

The Past ◽

Transmission Electron ◽

Organ Systems ◽

The Many

The role of electron microscopy and associated techniques is assured in diagnostic pathology. At the present time, most of the progress has been made on tissues examined by transmission electron microscopy (TEM) and correlated with light microscopy (LM) and by cytochemistry using both plastic and paraffin-embedded materials. As mentioned elsewhere in this symposium, this has revolutionized many fields of pathology including diagnostic, anatomic and clinical pathology. It began with the kidney; however, it has now been extended to most other organ systems and to tumor diagnosis in general. The results of the past few years tend to indicate the future directions and needs of this expanding field. Now, in addition to routine EM, pathologists have access to the many newly developed methods and instruments mentioned below which should aid considerably not only in diagnostic pathology but in investigative pathology as well.

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Transmission Electron Microscopy studies of texture of Cr underlayer of magnetic recording hard disk

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100087215 ◽

1991 ◽

Vol 49 ◽

pp. 580-581

Author(s):

L. Tang ◽

G. Thomas ◽

M. R. Khan ◽

S. L. Duan

Keyword(s):

Electron Microscopy ◽

Thin Films ◽

Transmission Electron Microscopy ◽

Magnetic Recording ◽

Magnetic Thin Films ◽

Magnetic Films ◽

Substrate Bias ◽

Epitaxial Relationship ◽

Transmission Electron

Cr thin films are often used as underlayers for Co alloy magnetic thin films, such as Co1, CoNi2, and CoNiCr3, for high density longitudinal magnetic recording. It is belived that the role of the Cr underlayer is to control the growth and texture of the Co alloy magnetic thin films, and, then, to increase the in plane coercivity of the films. Although many epitaxial relationship between the Cr underlayer and the magnetic films, such as ﹛1010﹜Co/ {110﹜Cr4, ﹛2110﹜Co/ ﹛001﹜Cr5, ﹛0002﹜Co/﹛110﹜Cr6, have been suggested and appear to be related to the Cr thickness, the texture of the Cr underlayer itself is still not understood very well. In this study, the texture of a 2000 Å thick Cr underlayer on Nip/Al substrate for thin films of (Co75Ni25)1-xTix dc-sputtered with - 200 V substrate bias is investigated by electron microscopy.

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Transmission Electron Microscopy of Evaporated Perylene Thin Films

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100135289 ◽

1990 ◽

Vol 48 (2) ◽

pp. 336-337

Author(s):

C. Ewins ◽

J.R. Fryer

Keyword(s):

Electron Microscopy ◽

Thin Films ◽

Transmission Electron Microscopy ◽

Molecular Electronics ◽

High Vacuum ◽

Amorphous Layer ◽

Electric Heater ◽

Transmission Electron ◽

Polycyclic Aromatic

The preparation of thin films of organic molecules is currently receiving much attention because of the need to produce good quality thin films for molecular electronics. We have produced thin films of the polycyclic aromatic, perylene C10H12 by evaporation under high vacuum onto a potassium chloride (KCl) substrate. The role of substrate temperature in determining the morphology and crystallography of the films was then investigated by transmission electron microscopy (TEM).The substrate studied was the (001) face of a freshly cleaved crystal of KCl. The temperature of the KCl was controlled by an electric heater or a cold finger. The KCl was heated to 200°C under a vacuum of 10-6 torr and allowed to cool to the desired temperature. The perylene was then evaporated over a period of one minute from a molybdenum boat at a distance of 10cm from the KCl. The perylene thin film was then backed with an amorphous layer of carbon and floated onto copper microscope grids.

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Fine structure and possible functions of the hermaphrodite duct of some pulmonate snails (Mollusca: Gastropoda)

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100157887 ◽

1990 ◽

Vol 48 (3) ◽

pp. 68-69

Author(s):

Alan N. Hodgson

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Fine Structure ◽

Seminal Vesicle ◽

Reproductive Biology ◽

Light Microscope ◽

Light Microscope Level ◽

Transmission Electron ◽

Standard Techniques

The hermaphrodite duct of pulmonate snails connects the ovotestis to the fertilization pouch. The duct is typically divided into three zones; aproximal duct which leaves the ovotestis, the middle duct (seminal vesicle) and the distal ovotestis duct. The seminal vesicle forms the major portion of the duct and is thought to store sperm prior to copulation. In addition the duct may also play a role in sperm maturation and degredation. Although the structure of the seminal vesicle has been described for a number of snails at the light microscope level there appear to be only two descriptions of the ultrastructure of this tissue. Clearly if the role of the hermaphrodite duct in the reproductive biology of pulmonatesis to be understood, knowledge of its fine structure is required.Hermaphrodite ducts, both containing and lacking sperm, of species of the terrestrial pulmonate genera Sphincterochila, Levantina, and Helix and the marine pulmonate genus Siphonaria were prepared for transmission electron microscopy by standard techniques.

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Fine Morphology of Antennal and Ovipositor Sensory Structures of the Gall Chestnut Wasp, Dryocosmus kuriphilus

Insects ◽

10.3390/insects12030231 ◽

2021 ◽

Vol 12 (3) ◽

pp. 231

Author(s):

Milos Sevarika ◽

Marco Valerio Rossi Stacconi ◽

Roberto Romani

Keyword(s):

Ultrastructural Organization ◽

Antennal Sensilla ◽

Dryocosmus Kuriphilus ◽

Transmission Electron ◽

Significant Damage ◽

Sensory Structures ◽

Host Plant Location ◽

Oviposition Sites ◽

Fine Morphology

Dryocosmus kuriphilus is a gall-inducing insect, which can cause significant damage on plants of the genus Castanea Mill., 1754. Antennae and ovipositor are the main sensory organs involved in the location of suitable oviposition sites. Antennal sensilla are involved in the host plant location, while ovipositor sensilla assess the suitability of the ovipositional bud. On both organs, diverse sensillar organs are present. Here, the distribution and ultrastructural organization of the sensilla were investigated by scanning and transmission electron microscopy. The antennae of D. kuriphilus are filiform and composed of 14 antennomeres, with the distal flagellomere bearing the highest number of sensilla. On the antennae, 6 sensilla types were found; sensilla chaetica, campaniformia, coeloconica-I, coeloconica-II, trichoidea and placoidea. The sensilla placoidea and trichoidea were the most abundant types. On the external walls of the ovipositor, gustatory and mechanoreceptive sensilla were observed. Internally, the egg channel hosted two additional sensory structures. The putative functional role of each sensilla in the context of insect’s ecology is discussed as well as the ovipositional mechanism used by this insect.

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Antioxidant and Signaling Role of Plastid-Derived Isoprenoid Quinones and Chromanols

International Journal of Molecular Sciences ◽

10.3390/ijms22062950 ◽

2021 ◽

Vol 22 (6) ◽

pp. 2950

Author(s):

Beatrycze Nowicka ◽

Agnieszka Trela-Makowej ◽

Dariusz Latowski ◽

Kazimierz Strzalka ◽

Renata Szymańska

Keyword(s):

Current Knowledge ◽

Stress Factors ◽

Oxygenic Photosynthesis ◽

Ros Generation ◽

Main Site ◽

Storage Lipids ◽

Abiotic Stress Factors ◽

Cell Components ◽

And Function

Plant prenyllipids, especially isoprenoid chromanols and quinols, are very efficient low-molecular-weight lipophilic antioxidants, protecting membranes and storage lipids from reactive oxygen species (ROS). ROS are byproducts of aerobic metabolism that can damage cell components, they are also known to play a role in signaling. Plants are particularly prone to oxidative damage because oxygenic photosynthesis results in O2 formation in their green tissues. In addition, the photosynthetic electron transfer chain is an important source of ROS. Therefore, chloroplasts are the main site of ROS generation in plant cells during the light reactions of photosynthesis, and plastidic antioxidants are crucial to prevent oxidative stress, which occurs when plants are exposed to various types of stress factors, both biotic and abiotic. The increase in antioxidant content during stress acclimation is a common phenomenon. In the present review, we describe the mechanisms of ROS (singlet oxygen, superoxide, hydrogen peroxide and hydroxyl radical) production in chloroplasts in general and during exposure to abiotic stress factors, such as high light, low temperature, drought and salinity. We highlight the dual role of their presence: negative (i.e., lipid peroxidation, pigment and protein oxidation) and positive (i.e., contribution in redox-based physiological processes). Then we provide a summary of current knowledge concerning plastidic prenyllipid antioxidants belonging to isoprenoid chromanols and quinols, as well as their structure, occurrence, biosynthesis and function both in ROS detoxification and signaling.

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3,4-dehydro-L-proline Induces Programmed Cell Death in the Roots of Brachypodium distachyon

International Journal of Molecular Sciences ◽

10.3390/ijms22147548 ◽

2021 ◽

Vol 22 (14) ◽

pp. 7548

Author(s):

Artur Pinski ◽

Alexander Betekhtin ◽

Jolanta Kwasniewska ◽

Lukasz Chajec ◽

Elzbieta Wolny ◽

...

Keyword(s):

Cell Death ◽

Brachypodium Distachyon ◽

Root Hairs ◽

Terminal Deoxynucleotidyl Transferase ◽

Cell Wall Proteins ◽

Root Epidermis ◽

Root Hair Development ◽

Transmission Electron ◽

Hair Development

As cell wall proteins, the hydroxyproline-rich glycoproteins (HRGPs) take part in plant growth and various developmental processes. To fulfil their functions, HRGPs, extensins (EXTs) in particular, undergo the hydroxylation of proline by the prolyl-4-hydroxylases. The activity of these enzymes can be inhibited with 3,4-dehydro-L-proline (3,4-DHP), which enables its application to reveal the functions of the HRGPs. Thus, to study the involvement of HRGPs in the development of root hairs and roots, we treated seedlings of Brachypodium distachyon with 250 µM, 500 µM, and 750 µM of 3,4-DHP. The histological observations showed that the root epidermis cells and the cortex cells beneath them ruptured. The immunostaining experiments using the JIM20 antibody, which recognizes the EXT epitopes, demonstrated the higher abundance of this epitope in the control compared to the treated samples. The transmission electron microscopy analyses revealed morphological and ultrastructural features that are typical for the vacuolar-type of cell death. Using the TUNEL test (terminal deoxynucleotidyl transferase dUTP nick end labelling), we showed an increase in the number of nuclei with damaged DNA in the roots that had been treated with 3,4-DHP compared to the control. Finally, an analysis of two metacaspases’ gene activity revealed an increase in their expression in the treated roots. Altogether, our results show that inhibiting the prolyl-4-hydroxylases with 3,4-DHP results in a vacuolar-type of cell death in roots, thereby highlighting the important role of HRGPs in root hair development and root growth.

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The Role of Microscopy in Preclinical Safety Assessment

Microscopy and Microanalysis ◽

10.1017/s143192760003748x ◽

2000 ◽

Vol 6 (S2) ◽

pp. 998-999

Author(s):

Barbara J. Dovey-Hartman

Keyword(s):

Electron Microscopy ◽

Vital Role ◽

Pathology Report ◽

Clinical Phase ◽

Transmission Electron ◽

New Chemical Entities ◽

Regulatory Submissions ◽

Hematoxylin And Eosin ◽

Preclinical Safety

Microscopy plays a vital role in assessing the safety of New Chemical Entities (NCE) in the pre-clinical phase of drug development. Light microscopy (LM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM) are used at the Schering-Plough Research Institute (SPRI) for evaluation of NCE. To support regulatory submissions, NCE are routinely tested in rodents in short-term studies such as one-month toxicity studies, and in longterm studies such as oncogenicity studies that may last 24 months. At the completion of a study, the animals are necropsied and the required tissues collected and stored in fixative. The tissues for LM are processed to slides and stained with Hematoxylin and Eosin (H&E). The information derived from the examination of these tissues by LM becomes an essential part of the pathology report. The LM examination of these tissues usually yields the information needed to either progress a NCE or otherwise deter or halt development.

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Role of Nanocrystallites of Al-Based Glasses and H2O2 in Degradation Azo Dyes

Materials ◽

10.3390/ma14010039 ◽

2020 ◽

Vol 14 (1) ◽

pp. 39

Author(s):

Qi Chen ◽

Zhicheng Yan ◽

Hao Zhang ◽

KiBuem Kim ◽

Weimin Wang

Keyword(s):

Metallic Glasses ◽

Azo Dye ◽

Amorphous Matrix ◽

Nano Scale ◽

Transmission Electron ◽

Melt Spun ◽

Degradation Ability ◽

Tem Transmission Electron Microscopy ◽

Dye Solutions

Al-based metallic glasses have a special atomic structure and should have a unique degradation ability in azo dye solutions. The Al88Ni9Y3 (Y3), Al85Ni9Y6 (Y6) and Al82Ni9Y9 (Y9) glassy ribbons are melt spun and used in degrading methyl orange (MO) azo dye solution with adding H2O2. With increasing cY, the as-spun ribbons have an increasing GFA (glass formability) and gradually decreased the degradation rate of MO solution. TEM (transmission electron microscopy) results show that the Y3 ribbon has nano-scale crystallites, which may form the channels to transport elements to the surface for degrading the MO solution. After adding H2O2, the degradation efficiency of Al-based glasses is improved and the Y6 ribbon has formed nano-scale crystallites embedded in the amorphous matrix and it has the largest improvement in MO solution degradation. These results indicate that forming nano-scale crystallites and adding H2O2 are effective methods to improve the degradation ability of Al-based glasses in azo dye solutions.

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