scholarly journals Confocal microscopy reveals alterations of thylakoids in Limnospira fusiformis during prophage induction

PROTOPLASMA ◽  
2021 ◽  
Author(s):  
Maryam Alsadat Zekri ◽  
Michael Schagerl ◽  
Johannes Schweichhart ◽  
Ingeborg Lang
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Flow Cytometry ◽  
3D Visualization ◽  
Laser Scanning Microscopy ◽  
Host Cells ◽  
Strong Increase ◽  
Prophage Induction ◽  
Intact Cells ◽  
Transmission Electron

AbstractThe alkaliphilic cyanobacterium Limnospira fusiformis is an integral part in food webs of tropical soda lakes. Recently, sudden breakdowns of Limnospira sp. blooms in their natural environment have been linked to cyanophage infections. We studied ultrastructural details and prophage components in the laboratory by means of confocal laser scanning microscopy (CLSM) and transmission electron microscopy (TEM). For a comparison at the subcellular level, we included transmission electron microscopy (TEM) material of infected cells collected during a field survey. Compared to TEM, CLSM has the advantage to rapidly providing results for whole, intact cells. Moreover, many cells can be studied at once. We chemically induced lysogenic cyanophages by means of mitomycin C (MMC) treatments and studied the ultrastructural alterations of host cells. In parallel, the number of cyanophages was obtained by flow cytometry. After treatment of the culture with MMC, flow cytometry showed a strong increase in viral counts, i.e., prophage induction. CLSM reflected the re-organization of L. fusiformis with remarkable alterations of thylakoid arrangements after prophage induction. Our study provides a first step towards 3D visualization of ultrastructure of cyanobacteria and showed the high potential of CLSM to investigate viral-mediated modifications in these groups.

Characterization of cellular development and fatty acid accumulation in thraustochytrid Aurantiochytrium strains of Taiwan

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Natarajan Velmurugan ◽  
Yesupatham Sathishkumar ◽  
Shashanka Sonowal ◽  
Ka-Lai Pang ◽  
Yang Soo Lee
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Flow Cytometry ◽  
Cell Growth ◽  
Intracellular Lipid ◽  
Fatty Acid Accumulation ◽  
Transmission Electron ◽  
Intracellular Lipid Accumulation ◽  
High Level

Abstract Long-chain saturated and polyunsaturated fatty acids of two new thraustochytrid isolates cultured from Taiwan mangroves, Aurantiochytrium sp. IMB169 and Aurantiochytrium sp. IMB171, were characterized through their cell growth and development in relation to their intracellular lipid accumulation using transmission electron microscopy. Flow cytometry in combination with the lipophilic fluorescent dye BODIPY 505/515 was used to stain and characterize intracellular lipid bodies in the two isolates. The transmission electron microscopy and flow cytometry analyses revealed a progressive accumulation of lipid products in IMB169 and IMB171. Further, selective BODIPY stained cells were successfully separated and enriched using flow cytometry at single cell level. Among the two isolates, IMB169 was found to produce a high level of docosahexaenoic acid. The qualitative and analytical results obtained using electron microscopy and flow cytometry studies were validated by gas chromatography (GC). In addition, a quantitative baseline was established using cell growth, flow cytometry and GC analyses for developing an efficient bioprocessing methodology to selectively enrich thraustochytrids phenotypes with desirable characteristics.

scholarly journals Endlicheria bracteolata (Meisn.) Essential Oil as a Weapon Against Leishmania amazonensis: In Vitro Assay

Molecules ◽  
2019 ◽  
Vol 24 (14) ◽  
pp. 2525 ◽  
Author(s):  
Mariana Margatto Rottini ◽  
Ana Claudia Fernandes Amaral ◽  
José Luiz Pinto Ferreira ◽  
Edinilze Souza Coelho Oliveira ◽  
Jefferson Rocha de Andrade Silva ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Flow Cytometry ◽  
Essential Oil ◽  
Cutaneous Leishmaniasis ◽  
Leishmania Amazonensis ◽  
Vitro Assay ◽  
Transmission Electron ◽  
Biological Potential ◽  
Intracellular Amastigote

The difficulties encountered and the numerous side effects present in the treatment of cutaneous leishmaniasis have encouraged the research for new compounds that can complement or replace existing treatment. The growing scientific interest in the study of plants, which are already used in folk remedies, has led our group to test Endlicheria bracteolata essential oil against Leishmania amazonensis. Several species of the Lauraceae family, or their compounds, have relevant antiprotozoal activities Therefore, the biological potential on L. amazonensis forms from the essential oil of Endlicheria bracteolata leaves was verified for the first time in that work. The antileishmanial activity was evaluated against promastigotes and intracellular amastigotes, and cytotoxicity were performed with J774.G8, which were incubated with different concentrations of E. bracteolata essential oil. Transmission electron microscopy and flow cytometry were performed with E. bracteolata essential oil IC50. Promastigote forms showed E. bracteolata essential oil IC50 of 7.945 ± 1.285 µg/mL (24 h) and 6.186 ± 1.226 µg/mL (48 h), while for intracellular amastigote forms it was 3.546 ± 1.184 µg/mL (24 h). The CC50 was 15.14 ± 0.090 µg/mL showing that E. bracteolata essential oil is less toxic to macrophages than to parasites. Transmission electron microscopy showed that E. bracteolata essential oil treatment is capable of inducing mitochondrial damage to promastigote and intracellular amastigote forms, while flow cytometry showed ΔѰm disruption in treated parasites. These results could bring about new possibilities to develop products based on E. bracteolata essential oil to treat cutaneous leishmaniasis, especially for people who cannot receive the conventional therapy.

Microtubule targeted therapeutics loaded polymeric assembled nanospheres for potentiation of antineoplastic activity

2016 ◽  
Vol 186 ◽  
pp. 45-59 ◽  
Author(s):  
Radhika Poojari ◽  
Rohit Srivastava ◽  
Dulal Panda
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Laser Scanning ◽  
Present Approach ◽  
Laser Scanning Microscopy ◽  
Microtubule Assembly ◽  
Confocal Laser ◽  
Cancer Cell Proliferation ◽  
Transmission Electron ◽  
Hepatocarcinoma Cells

Polymeric nanoassemblies represent an attractive strategy for efficient cellular internalization of microtubule targeted anticancer drugs. Using dynamic light scattering, zeta potential, transmission electron microscopy and scanning electron microscopy, the physical properties and surface morphology of microtubule-binding PEGylated PLGA assembled nanospheres (100–200 nm) were analyzed. The present approach leads to strong internalization as observed by confocal laser scanning microscopy and transmission electron microscopy in hepatocarcinoma cells. The effect of these nanoassemblies on microtubules and mitosis were explored using immunofluorescence microscopy. The effects of these nanoassemblies on cancer cell proliferation and cell death revealed their antitumor enhancing effects. Perturbation of the microtubule assembly, mitosis and nuclear modulations potentiated the antineoplastic effects delivered via nanospheres in hepatocarcinoma cells. The extensive biomolecular and physical characterizations of the synthesized nanoassemblies will help to design potent therapeutic materials and the present approach can be applied to deliver microtubule-targeted drugs for liver cancer therapy.

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