scholarly journals Human microsporidian pathogen Encephalitozoon intestinalis impinges on enterocyte membrane trafficking and signaling

2021 ◽  
pp. jcs.253757
Author(s):  
Juan Flores ◽  
Peter M. Takvorian ◽  
Louis M. Weiss ◽  
Ann Cali ◽  
Nan Gao

Microsporidia are a large phylum of obligate intracellular parasites. Approximately a dozen species of microsporidia infect humans where they are responsible for a variety of diseases and occasionally death, especially in immunocompromised individuals. To better understand the impact of microsporidia on human cells, we infected human colonic Caco2 cells with Encephalitozoon intestinalis, and showed that these enterocyte cultures can be used to recapitulate the parasites’ life cycle, including the spread of infection with infective spores. Using transmission electron microscopy, we describe this lifecycle and demonstrate nuclear, mitochondrial, and microvillar alterations by this pathogen. We also analyzed the transcriptome of infected cells to reveal host cell signaling alterations upon infection. These high-resolution imaging and transcriptional profiling analysis shed light on the impact of the microsporidial infection on its primary human target cell type.

Author(s):  
J. W. Horn ◽  
B. J. Dovey-Hartman ◽  
V. P. Meador

Osmium tetroxide (OsO4) is a universally used secondary fixative for routine transmission electron microscopic evaluation of biological specimens. Use of OsO4 results in good ultrastructural preservation and electron density but several factors, such as concentration, length of exposure, and temperature, impact overall results. Potassium ferricyanide, an additive used primarily in combination with OsO4, has mainly been used to enhance the contrast of lipids, glycogen, cell membranes, and membranous organelles. The purpose of this project was to compare the secondary fixative solutions, OsO4 vs. OsO4 with potassium ferricyanide, and secondary fixative temperature for determining which combination gives optimal ultrastructural fixation and enhanced organelle staining/contrast.Fresh rat liver samples were diced to ∼1 mm3 blocks, placed into porous processing capsules/baskets, preserved in buffered 2% formaldehyde/2.5% glutaraldehyde solution, and rinsed with 0.12 M cacodylate buffer (pH 7.2). Tissue processing capsules were separated (3 capsules/secondary fixative.solution) and secondarily fixed (table) for 90 minutes. Tissues were buffer rinsed, dehydrated with ascending concentrations of ethanol solutions, infiltrated, and embedded in epoxy resin.


Author(s):  
Robert C. Cieslinski ◽  
H. Craig Silvis ◽  
Daniel J. Murray

An understanding of the mechanical behavior polymers in the ductile-brittle transition region will result in materials with improved properties. A technique has been developed that allows the realtime observation of dynamic plane stress failure mechanisms in the transmission electron microscope. With the addition of a cryo-tensile stage, this technique has been extented to -173°C, allowing the observation of deformation during the ductile-brittle transition.The technique makes use of an annealed copper cartridge in which a thin section of bulk polymer specimen is bonded and plastically deformed in tension in the TEM using a screw-driven tensile stage. In contrast to previous deformation studies on solvent-cast films, this technique can examine the frozen-in morphology of a molded part.The deformation behavior of polypropylene and polypropylene impact modified with EPDM (ethylene-propylene diene modified) and PE (polyethylene) rubbers were investigated as function of temperature and the molecular weight of the impact modifier.


Author(s):  
Terrence Reilly ◽  
Al Pelillo ◽  
Barbara Miner

The use of transmission electron microscopes (TEM) has proven to be very valuable in the observation of semiconductor devices. The need for high resolution imaging becomes more important as the devices become smaller and more complex. However, the sample preparation for TEM observation of semiconductor devices have generally proven to be complex and time consuming. The use of ion milling machines usually require a certain degree of expertise and allow a very limited viewing area. Recently, the use of an ultra high resolution "immersion lens" cold cathode field emission scanning electron microscope (CFESEM) has proven to be very useful in the observation of semiconductor devices. Particularly at low accelerating voltages where compositional contrast is increased. The Hitachi S-900 has provided comparable resolution to a 300kV TEM on semiconductor cross sections. Using the CFESEM to supplement work currently being done with high voltage TEMs provides many advantages: sample preparation time is greatly reduced and the observation area has also been increased to 7mm. The larger viewing area provides the operator a much greater area to search for a particular feature of interest. More samples can be imaged on the CFESEM, leaving the TEM for analyses requiring diffraction work and/or detecting the nature of the crystallinity.


2019 ◽  
Vol 25 (6) ◽  
pp. 1466-1470 ◽  
Author(s):  
Rituparna Chatterjee ◽  
Subhajit Saha ◽  
Karamjyoti Panigrahi ◽  
Uttam Kumar Ghorai ◽  
Gopes Chandra Das ◽  
...  

AbstractIn this work, strongly blue emitting Ce3+-activated BaAl2O4 nanophosphors were successfully synthesized by a sol–gel technique. The crystal structure, morphology, and microstructure of the nanophosphors have been studied by X-ray powder diffraction, field emission scanning electron microscopy, and high-resolution transmission electron microscopy. The photoluminescence spectra show the impact of concentration variation of Ce3+ on the photoluminescence emission of the phosphor. These nanophosphors display intense blue emission peaking at 422 nm generated by the Ce3+ 5d → 4f transition under 350 nm excitation. Our results reveal that this nanophosphor has the capability to take part in the emergent domain of solid-state lighting and field-emission display devices.


Plants ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 583
Author(s):  
Reda E. Abdelhameed ◽  
Nagwa I. Abu-Elsaad ◽  
Arafat Abdel Hamed Abdel Latef ◽  
Rabab A. Metwally

Important gaps in knowledge remain regarding the potential of nanoparticles (NPs) for plants, particularly the existence of helpful microorganisms, for instance, arbuscular mycorrhizal (AM) fungi present in the soil. Hence, more profound studies are required to distinguish the impact of NPs on plant growth inoculated with AM fungi and their role in NP uptake to develop smart nanotechnology implementations in crop improvement. Zinc ferrite (ZnFe2O4) NPs are prepared via the citrate technique and defined by X-ray diffraction (XRD) as well as transmission electron microscopy for several physical properties. The analysis of the XRD pattern confirmed the creation of a nanocrystalline structure with a crystallite size equal to 25.4 nm. The effects of ZnFe2O4 NP on AM fungi, growth and pigment content as well as nutrient uptake of pea (Pisum sativum) plants were assessed. ZnFe2O4 NP application caused a slight decrease in root colonization. However, its application showed an augmentation of 74.36% and 91.89% in AM pea plant shoots and roots’ fresh weights, respectively, compared to the control. Moreover, the synthesized ZnFe2O4 NP uptake by plant roots and their contents were enhanced by AM fungi. These findings suggest the safe use of ZnFe2O4 NPs in nano-agricultural applications for plant development with AM fungi.


BMJ Open ◽  
2020 ◽  
Vol 10 (10) ◽  
pp. e043010
Author(s):  
Jane Lyons ◽  
Ashley Akbari ◽  
Fatemeh Torabi ◽  
Gareth I Davies ◽  
Laura North ◽  
...  

IntroductionThe emergence of the novel respiratory SARS-CoV-2 and subsequent COVID-19 pandemic have required rapid assimilation of population-level data to understand and control the spread of infection in the general and vulnerable populations. Rapid analyses are needed to inform policy development and target interventions to at-risk groups to prevent serious health outcomes. We aim to provide an accessible research platform to determine demographic, socioeconomic and clinical risk factors for infection, morbidity and mortality of COVID-19, to measure the impact of COVID-19 on healthcare utilisation and long-term health, and to enable the evaluation of natural experiments of policy interventions.Methods and analysisTwo privacy-protecting population-level cohorts have been created and derived from multisourced demographic and healthcare data. The C20 cohort consists of 3.2 million people in Wales on the 1 January 2020 with follow-up until 31 May 2020. The complete cohort dataset will be updated monthly with some individual datasets available daily. The C16 cohort consists of 3 million people in Wales on the 1 January 2016 with follow-up to 31 December 2019. C16 is designed as a counterfactual cohort to provide contextual comparative population data on disease, health service utilisation and mortality. Study outcomes will: (a) characterise the epidemiology of COVID-19, (b) assess socioeconomic and demographic influences on infection and outcomes, (c) measure the impact of COVID-19 on short -term and longer-term population outcomes and (d) undertake studies on the transmission and spatial spread of infection.Ethics and disseminationThe Secure Anonymised Information Linkage-independent Information Governance Review Panel has approved this study. The study findings will be presented to policy groups, public meetings, national and international conferences, and published in peer-reviewed journals.


Animals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1414
Author(s):  
Josep M. Cambra ◽  
Emilio A. Martinez ◽  
Heriberto Rodriguez-Martinez ◽  
Maria A. Gil ◽  
Cristina Cuello

The development of chemically defined media is a growing trend in in vitro embryo production (IVP). Recently, traditional undefined culture medium with bovine serum albumin (BSA) has been successfully replaced by a chemically defined medium using substances with embryotrophic properties such as platelet factor 4 (PF4). Although the use of this medium sustains IVP, the impact of defined media on the embryonic transcriptome has not been fully elucidated. This study analyzed the transcriptome of porcine IVP blastocysts, cultured in defined (PF4 group) and undefined media (BSA group) by microarrays. In vivo-derived blastocysts (IVV group) were used as a standard of maximum embryo quality. The results showed no differentially expressed genes (DEG) between the PF4 and BSA groups. However, a total of 2780 and 2577 DEGs were detected when comparing the PF4 or the BSA group with the IVV group, respectively. Most of these genes were common in both in vitro groups (2132) and present in some enriched pathways, such as cell cycle, lysosome and/or metabolic pathways. These results show that IVP conditions strongly affect embryo transcriptome and that the defined culture medium with PF4 is a guaranteed replacement for traditional culture with BSA.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mikolaj Grabowski ◽  
Ewa Grzanka ◽  
Szymon Grzanka ◽  
Artur Lachowski ◽  
Julita Smalc-Koziorowska ◽  
...  

AbstractThe aim of this paper is to give an experimental evidence that point defects (most probably gallium vacancies) induce decomposition of InGaN quantum wells (QWs) at high temperatures. In the experiment performed, we implanted GaN:Si/sapphire substrates with helium ions in order to introduce a high density of point defects. Then, we grew InGaN QWs on such substrates at temperature of 730 °C, what caused elimination of most (but not all) of the implantation-induced point defects expanding the crystal lattice. The InGaN QWs were almost identical to those grown on unimplanted GaN substrates. In the next step of the experiment, we annealed samples grown on unimplanted and implanted GaN at temperatures of 900 °C, 920 °C and 940 °C for half an hour. The samples were examined using Photoluminescence, X-ray Diffraction and Transmission Electron Microscopy. We found out that the decomposition of InGaN QWs started at lower temperatures for the samples grown on the implanted GaN substrates what provides a strong experimental support that point defects play important role in InGaN decomposition at high temperatures.


Nanomaterials ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1929
Author(s):  
Tereza Cervena ◽  
Andrea Rossnerova ◽  
Tana Zavodna ◽  
Jitka Sikorova ◽  
Kristyna Vrbova ◽  
...  

The evaluation of the frequency of micronuclei (MN) is a broadly utilised approach in in vitro toxicity testing. Nevertheless, the specific properties of nanomaterials (NMs) give rise to concerns regarding the optimal methodological variants of the MN assay. In bronchial epithelial cells (BEAS-2B), we tested the genotoxicity of five types of NMs (TiO2: NM101, NM103; SiO2: NM200; Ag: NM300K, NM302) using four variants of MN protocols, differing in the time of exposure and the application of cytochalasin-B combined with the simultaneous and delayed co-treatment with NMs. Using transmission electron microscopy, we evaluated the impact of cytochalasin-B on the transport of NMs into the cells. To assess the behaviour of NMs in a culture media for individual testing conditions, we used dynamic light scattering measurement. The presence of NMs in the cells, their intracellular aggregation and dispersion properties were comparable when tests with or without cytochalasin-B were performed. The genotoxic potential of various TiO2 and Ag particles differed (NM101 < NM103 and NM302 < NM300K, respectively). The application of cytochalasin-B tended to increase the percentage of aberrant cells. In conclusion, the comparison of the testing strategies revealed that the level of DNA damage induced by NMs is affected by the selected methodological approach. This fact should be considered in the interpretation of the results of genotoxicity tests.


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