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1.
Nature ; 594(7862): 201-206, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-34108694

RESUMO

The performance of light microscopes is limited by the stochastic nature of light, which exists in discrete packets of energy known as photons. Randomness in the times that photons are detected introduces shot noise, which fundamentally constrains sensitivity, resolution and speed1. Although the long-established solution to this problem is to increase the intensity of the illumination light, this is not always possible when investigating living systems, because bright lasers can severely disturb biological processes2-4. Theory predicts that biological imaging may be improved without increasing light intensity by using quantum photon correlations1,5. Here we experimentally show that quantum correlations allow a signal-to-noise ratio beyond the photodamage limit of conventional microscopy. Our microscope is a coherent Raman microscope that offers subwavelength resolution and incorporates bright quantum correlated illumination. The correlations allow imaging of molecular bonds within a cell with a 35 per cent improved signal-to-noise ratio compared with conventional microscopy, corresponding to a 14 per cent improvement in concentration sensitivity. This enables the observation of biological structures that would not otherwise be resolved. Coherent Raman microscopes allow highly selective biomolecular fingerprinting in unlabelled specimens6,7, but photodamage is a major roadblock for many applications8,9. By showing that the photodamage limit can be overcome, our work will enable order-of-magnitude improvements in the signal-to-noise ratio and the imaging speed.


Assuntos
Lasers , Iluminação , Microscopia/métodos , Fótons , Teoria Quântica , Análise Espectral Raman , Células/patologia , Células/efeitos da radiação , Lasers/efeitos adversos , Iluminação/efeitos adversos , Microscopia/instrumentação , Fótons/efeitos adversos , Razão Sinal-Ruído , Análise Espectral Raman/instrumentação , Análise Espectral Raman/métodos
2.
J Vis Exp ; (168)2021 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-33682854

RESUMO

The importance of dosimetry protocols and standards for radiobiological studies is self-evident. Several protocols have been proposed for dose determination using low energy X-ray facilities, but depending on the irradiation configurations, samples, materials or beam quality, it is sometimes difficult to know which protocol is the most appropriate to employ. We, therefore, propose a dosimetry protocol for cell irradiations using low energy X-ray facility. The aim of this method is to perform the dose estimation at the level of the cell monolayer to make it as close as possible to real cell irradiation conditions. The different steps of the protocol are as follows: determination of the irradiation parameters (high voltage, intensity, cell container etc.), determination of the beam quality index (high voltage-half value layer couple), dose rate measurement with ionization chamber calibrated in air kerma conditions, quantification of the attenuation and scattering of the cell culture medium with EBT3 radiochromic films, and determination of the dose rate at the cellular level. This methodology must be performed for each new cell irradiation configuration as the modification of only one parameter can strongly impact the real dose deposition at the level of the cell monolayer, particularly involving low energy X-rays.


Assuntos
Células/efeitos da radiação , Radiometria , Calibragem , Simulação por Computador , Meios de Cultura , Relação Dose-Resposta à Radiação , Raios X
3.
Adv Exp Med Biol ; 1293: 209-224, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33398815

RESUMO

Optogenetic approaches combine the power to allocate optogenetic tools (proteins) to specific cell populations (defined genetically or functionally) and the use of light-based interfaces between biological wetware (cells and tissues) and hardware (controllers and recorders). The optogenetic toolbox contains two main compartments: tools to interfere with cellular processes and tools to monitor cellular events. Among the latter are genetically encoded voltage indicators (GEVIs). This chapter outlines the development, current state of the art and prospects of emerging optical GEVI imaging technologies.


Assuntos
Proteínas Luminescentes/genética , Potenciais da Membrana , Optogenética/métodos , Células/metabolismo , Células/efeitos da radiação , Potenciais da Membrana/efeitos da radiação , Optogenética/instrumentação
4.
Int J Mol Sci ; 21(12)2020 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-32580352

RESUMO

Gold nanoparticle (GNP) enhanced proton therapy is a promising treatment concept offering increased therapeutic effect. It has been demonstrated in experiments which provided indications that reactive species play a major role. Simulations of the radiolysis yield from GNPs within a cell model were performed using the Geant4 toolkit. The effect of GNP cluster size, distribution and number, cell and nuclear membrane absorption and intercellular yields were evaluated. It was found that clusters distributed near the nucleus increased the nucleus yield by 91% while reducing the cytoplasm yield by 7% relative to a disperse distribution. Smaller cluster sizes increased the yield, 200 nm clusters had nucleus and cytoplasm yields 117% and 35% greater than 500 nm clusters. Nuclear membrane absorption reduced the cytoplasm and nucleus yields by 8% and 35% respectively to a permeable membrane. Intercellular enhancement was negligible. Smaller GNP clusters delivered near sub-cellular targets maximise radiosensitisation. Nuclear membrane absorption reduces the nucleus yield, but can damage the membrane providing another potential pathway for biological effect. The minimal effect on adjacent cells demonstrates that GNPs provide a targeted enhancement for proton therapy, only effecting cells with GNPs internalised. The provided quantitative data will aid further experiments and clinical trials.


Assuntos
Células/efeitos da radiação , Ouro/química , Nanopartículas Metálicas/química , Modelos Biológicos , Terapia com Prótons , Radiólise de Impulso , Radiossensibilizantes/química , Método de Monte Carlo
5.
Cell ; 179(5): 1098-1111.e23, 2019 11 14.
Artigo em Inglês | MEDLINE | ID: mdl-31730852

RESUMO

We report a 100-million atom-scale model of an entire cell organelle, a photosynthetic chromatophore vesicle from a purple bacterium, that reveals the cascade of energy conversion steps culminating in the generation of ATP from sunlight. Molecular dynamics simulations of this vesicle elucidate how the integral membrane complexes influence local curvature to tune photoexcitation of pigments. Brownian dynamics of small molecules within the chromatophore probe the mechanisms of directional charge transport under various pH and salinity conditions. Reproducing phenotypic properties from atomistic details, a kinetic model evinces that low-light adaptations of the bacterium emerge as a spontaneous outcome of optimizing the balance between the chromatophore's structural integrity and robust energy conversion. Parallels are drawn with the more universal mitochondrial bioenergetic machinery, from whence molecular-scale insights into the mechanism of cellular aging are inferred. Together, our integrative method and spectroscopic experiments pave the way to first-principles modeling of whole living cells.


Assuntos
Células/metabolismo , Metabolismo Energético , Adaptação Fisiológica/efeitos da radiação , Trifosfato de Adenosina/metabolismo , Benzoquinonas/metabolismo , Membrana Celular/metabolismo , Membrana Celular/efeitos da radiação , Células/efeitos da radiação , Cromatóforos/metabolismo , Citocromos c2/metabolismo , Difusão , Transporte de Elétrons/efeitos da radiação , Metabolismo Energético/efeitos da radiação , Meio Ambiente , Ligação de Hidrogênio , Cinética , Luz , Simulação de Dinâmica Molecular , Fenótipo , Proteínas/metabolismo , Rhodobacter sphaeroides/fisiologia , Rhodobacter sphaeroides/efeitos da radiação , Eletricidade Estática , Estresse Fisiológico/efeitos da radiação , Temperatura
6.
Electromagn Biol Med ; 38(4): 231-248, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31450976

RESUMO

This paper summarizes studies on changes in cellular free radical activities from exposure to static and extremely-low frequency (ELF) electromagnetic fields (EMF), particularly magnetic fields. Changes in free radical activities, including levels of cellular reactive oxygen (ROS)/nitrogen (RNS) species and endogenous antioxidant enzymes and compounds that maintain physiological free radical concentrations in cells, is one of the most consistent effects of EMF exposure. These changes have been reported to affect many physiological functions such as DNA damage; immune response; inflammatory response; cell proliferation and differentiation; wound healing; neural electrical activities; and behavior. An important consideration is the effects of EMF-induced changes in free radicals on cell proliferation and differentiation. These cellular processes could affect cancer development and proper growth and development in organisms. On the other hand, they could cause selective killing of cancer cells, for instance, via the generation of the highly cytotoxic hydroxyl free radical by the Fenton Reaction. This provides a possibility of using these electromagnetic fields as a non-invasive and low side-effect cancer therapy. Static- and ELF-EMF probably play important roles in the evolution of living organisms. They are cues used in many critical survival functions, such as foraging, migration, and reproduction. Living organisms can detect and respond immediately to low environmental levels of these fields. Free radical processes are involved in some of these mechanisms. At this time, there is no credible hypothesis or mechanism that can adequately explain all the observed effects of static- and ELF-EMF on free radical processes. We are actually at the impasse that there are more questions than answers.


Assuntos
Células/metabolismo , Células/efeitos da radiação , Campos Eletromagnéticos , Radicais Livres/metabolismo , Animais , Células/citologia , Evolução Molecular , Humanos , Peróxido de Hidrogênio/metabolismo , Ferro/metabolismo
7.
Phys Chem Chem Phys ; 21(29): 15917-15931, 2019 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-31309206

RESUMO

X-ray nanochemistry studies how to use nanomaterials and particularly how to create new nanomaterials to increase the effects of X-rays such as chemical reactivity, damage to cells, tumor destruction, scintillation and more. The increase, also called enhancement, can be categorized into several groups, and the current categorization of enhancement follows a natural division of physical, chemical and biological enhancement based on how nanomaterials behave under X-ray irradiation. In physical enhancement, electrons released from atoms in the nanomaterials upon X-ray ionization interact with the nanomaterials and surrounding media to increase the effects. Scintillation also belongs to this category. Chemical enhancement results when reactive oxygen species (ROS) or reactive radical intermediates (RRI) produced in aqueous solutions under X-ray irradiation interact with the surface of catalytic nanomaterials to increase the effects. When the damage of cells is enhanced through biological pathways beyond the abovementioned physical or chemical enhancement due to the presence of nanomaterials under X-ray irradiation, the enhancement is called biological enhancement. Works supporting this systematic categorization, the reported values of these enhancements, and important aspects of the development of enhancement in the X-ray nanochemistry framework are given and discussed in this perspective.


Assuntos
Técnicas de Química Analítica/métodos , Nanoestruturas/química , Raios X , Células/química , Células/efeitos da radiação , Espécies Reativas de Oxigênio
8.
Mar Drugs ; 17(6)2019 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-31146377

RESUMO

Due to its special aromatic structure, isorenieratene is thought to be an active natural antioxidant and photo/UV damage inhibitor. In this work, isorenieratene that was extracted from Rhodococcus sp. B7740 isolated from the Arctic Ocean, showed excellent scavenging ability of both singlet oxygen and hydroxyl radical in the UVB-induced auto-oxidation process using the EPR method. Within an ARPE-19 cell model damaged by UVB radiation, isorenieratene showed fine protective effects (1.13 ± 0.03 fold) compared with macular xanthophylls (MXs) through upregulating of tspo. The molecular docking was firstly performed to investigate the interaction of isorenieratene with TSPO as a special ligand. Results showed isorenieratene might form a better binding conformation (S-score -8.5438) than MXs and indicate that isorenieratene not only can function as a direct antioxidant but also activate tspo in ARPE-19 cells. Thus, isorenieratene might ease the UV-related damages including age-related macular degeneration (AMD).


Assuntos
Carotenoides/farmacologia , Células/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Fenóis/farmacologia , Receptores de GABA/metabolismo , Rhodococcus/química , Animais , Regiões Árticas , Carotenoides/isolamento & purificação , Linhagem Celular , Células/efeitos da radiação , Citoproteção/efeitos dos fármacos , Humanos , Ligantes , Camundongos , Modelos Moleculares , Oceanos e Mares , Estresse Oxidativo/efeitos da radiação , Fenóis/isolamento & purificação , Extratos Vegetais/farmacologia , Estrutura Terciária de Proteína , Protetores contra Radiação/química , Protetores contra Radiação/farmacologia , Raios Ultravioleta/efeitos adversos
10.
PLoS One ; 14(3): e0213286, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30870450

RESUMO

Recent experiments have reported an effect of weak radiofrequency magnetic fields in the MHz-range on the concentrations of reactive oxygen species (ROS) in living cells. Since the energy that could possibly be deposited by the radiation is orders of magnitude smaller than the energy of molecular thermal motion, it was suggested that the effect was caused by the interaction of RF magnetic fields with transient radical pairs within the cells, affecting the ROS formation rates through the radical pair mechanism. It is, however, at present not entirely clear how to predict RF magnetic field effects at certain field frequency and intensity in nanoscale biomolecular systems. We suggest a possible recipe for interpreting the radiofrequency effects in cells by presenting a general workflow for calculation of the reactive perturbations inside a cell as a function of RF magnetic field strength and frequency. To justify the workflow, we discuss the effects of radiofrequency magnetic fields on generic spin systems to particularly illustrate how the reactive radicals could be affected by specific parameters of the experiment. We finally argue that the suggested workflow can be used to predict effects of radiofrequency magnetic fields on radical pairs in biological cells, which is specially important for wireless recharging technologies where one has to know of any harmful effects that exposure to such radiation might cause.


Assuntos
Células/efeitos da radiação , Campos Eletromagnéticos/efeitos adversos , Modelos Teóricos , Lesões por Radiação/etiologia , Espécies Reativas de Oxigênio/metabolismo , Células Cultivadas , Humanos
11.
Life Sci Space Res (Amst) ; 19: 68-75, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30482285

RESUMO

One of the specific properties of laser-driven radiation is a broadband energy spectrum, which is also a feature of the space radiation fields. This property can be used in materials science studies or radiobiology experiments to simulate the energy spectrum of space radiation exposures in a ground-based laboratory. However, the differences in effects between the higher dose rates of laser generated radiation and the lower dose rates of space radiation have to be investigated in separate, prior studies. A design for a high-throughput irradiation experiment and the associated Monte Carlo dose calculations for a broadband energy proton beam depositing energy in a cell monolayer is presented. Dose control and dose uniformity in the cell monolayer was achieved in the simulations using a variable thickness Ni attenuator. A set of target doses from 0.2 Gy to 4 Gy was obtained and dose uniformity was optimized to less than 4% variability. This work opens the possibility of single or multiple exposures, controllable, high-throughput irradiation experiments on biological samples or materials, using broadband energy particle beams generated by lasers, with relevance for space applications.


Assuntos
Células/efeitos da radiação , Radiometria/métodos , Voo Espacial , Células/citologia , Células Cultivadas , Humanos , Lasers , Método de Monte Carlo , Aceleradores de Partículas , Prótons , Doses de Radiação
12.
Langmuir ; 34(48): 14459-14471, 2018 12 04.
Artigo em Inglês | MEDLINE | ID: mdl-30392367

RESUMO

Optoregulated biointerfaces offer the possibility to manipulate the interactions between cell membrane receptors and the extracellular space. This Invited Feature Article summarizes recent efforts by our group and others during the past decade to develop light-responsive biointerfaces to stimulate cells and elicit cellular responses using photocleavable protecting groups (PPG) as our working tool. This article begins by providing a brief introduction to available PPGs, with a special focus on the widely used o-nitrobenzyl family, followed by an overview of molecular design principles for the control of bioactivity in the context of cell-material interactions and the characterization methods to use in following the photoreaction at surfaces. We present various light-guided cellular processes using PPGs, including cell adhesion, release, migration, proliferation, and differentiation, both in vitro and in vivo. Finally, this Invited Feature Article closes with our perspective on the current status and future challenges of this topic.


Assuntos
Células/efeitos da radiação , Luz , Materiais Biocompatíveis/química , Materiais Biocompatíveis/metabolismo , Adesão Celular/efeitos da radiação , Diferenciação Celular/efeitos da radiação , Células/citologia , Células/metabolismo , Humanos , Hidrogéis/química , Hidrogéis/metabolismo
13.
Sensors (Basel) ; 18(7)2018 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-30002282

RESUMO

Graphene provides a unique way of sensing the local pH level of substances on the micrometric scale, with important implications for the monitoring of cellular metabolic activities where proton excretion could occur. Accordingly, an innovative biosensing approach for the quantification of the pH value of biological fluids, to be used also with small amounts of fluids, was realized and tested. It is based on the use of micro-Raman spectroscopy to detect the modifications of the graphene doping level induced by the contact of the graphene with the selected fluids. The approach was preliminarily tested on aqueous solutions of known pH values. It was then used to quantify the pH values of cell culture media directly exposed to different doses of X-ray radiation and to media exposed to X-ray-irradiated cells. The Raman response of cells placed on graphene layers was also examined.


Assuntos
Células/química , Células/efeitos da radiação , Meios de Cultura/química , Meios de Cultura/efeitos da radiação , Grafite/química , Análise Espectral Raman/métodos , Raios X , Humanos , Concentração de Íons de Hidrogênio
14.
PLoS One ; 13(5): e0195479, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29718923

RESUMO

Laser-induced cell transfer has been developed in recent years for the flexible and gentle printing of cells. Because of the high transfer rates and the superior cell survival rates, this technique has great potential for tissue engineering applications. However, the fact that material from an inorganic sacrificial layer, which is required for laser energy absorption, is usually transferred to the printed target structure, constitutes a major drawback of laser based cell printing. Therefore alternative approaches using deep UV laser sources and protein based acceptor films for energy absorption, have been introduced. Nevertheless, deep UV radiation can introduce DNA double strand breaks, thereby imposing the risk of carcinogenesis. Here we present a method for the laser-induced transfer of hydrogels and mammalian cells, which neither requires any sacrificial material for energy absorption, nor the use of UV lasers. Instead, we focus a near infrared femtosecond (fs) laser pulse (λ = 1030 nm, 450 fs) directly underneath a thin cell layer, suspended on top of a hydrogel reservoir, to induce a rapidly expanding cavitation bubble in the gel, which generates a jet of material, transferring cells and hydrogel from the gel/cell reservoir to an acceptor stage. By controlling laser pulse energy, well-defined cell-laden droplets can be transferred with high spatial resolution. The transferred human (SCP1) and murine (B16F1) cells show high survival rates, and good cell viability. Time laps microscopy reveals unaffected cell behavior including normal cell proliferation.


Assuntos
Células/citologia , Células/efeitos da radiação , Raios Infravermelhos , Lasers , Animais , Linhagem Celular Tumoral , Proliferação de Células , Sobrevivência Celular , Humanos , Camundongos , Fatores de Tempo
15.
Radiat Res ; 190(1): 5-11, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29697303

RESUMO

Aside from the generally accepted potential to cause DNA damage, it is becoming increasingly recognized that ionizing radiation has the capability to target the cellular epigenome. Epigenetics unifies the chemical marks and molecules that collectively facilitate the proper reading of genetic material. Among the epigenetic mechanisms of regulation, methylation of DNA is known to be the key player in the postirradiation response by controlling the expression of genetic information and activity of transposable elements. Radiation-induced alterations to DNA methylation may lead to cellular epigenetic reprogramming that, in turn, can substantially compromise the genomic integrity and has been proposed as one of the mechanisms of radiation-induced carcinogenesis. DNA methylation is strongly dependent on the one-carbon metabolism. This metabolic pathway is central to the support of DNA methylation by means of providing the donor of methyl groups, as well as for the synthesis of amino acids. To better understand the mechanisms of radiation-induced health effects, we study how exposure to radiation affects DNA methylation and one-carbon metabolism. Also, a tight interaction that exists between DNA methylation and one-carbon metabolism allows us to simultaneously manipulate both cellular epigenetic and metabolic profiles to modulate the normal and cancerous tissue response to radiotherapy.


Assuntos
Células/metabolismo , Células/efeitos da radiação , Epigênese Genética/efeitos da radiação , Animais , DNA/genética , Metilação de DNA/efeitos da radiação , Relação Dose-Resposta à Radiação , Humanos , Metabolômica , Neoplasias/genética , Neoplasias/patologia
16.
Artigo em Inglês | MEDLINE | ID: mdl-29148897

RESUMO

The exact mechanism that could explain the effects of radiofrequency (RF) radiation exposure at non-thermal level is still unknown. Increasing evidence suggests a possible involvement of reactive oxygen species (ROS) and development of oxidative stress. To test the proposed hypothesis, human neuroblastoma cells (SH-SY5Y) were exposed to 1800 MHz short-term RF exposure for 10, 30 and 60 minutes. Electric field strength within Gigahertz Transverse Electromagnetic cell (GTEM) was 30 V m-1 and specific absorption rate (SAR) was calculated to be 1.6 W kg-1. Cellular viability was measured by MTT assay and level of ROS was determined by fluorescent probe 2',7'-dichlorofluorescin diacetate. Concentrations of malondialdehyde and protein carbonyls were used to assess lipid and protein oxidative damage and antioxidant activity was evaluated by measuring concentrations of total glutathione (GSH). After radiation exposure, viability of irradiated cells remained within normal physiological values. Significantly higher ROS level was observed for every radiation exposure time. After 60 min of exposure, the applied radiation caused significant lipid and protein damage. The highest GSH concentration was detected after 10 minute-exposure. The results of our study showed enhanced susceptibility of SH-SY5Y cells for development of oxidative stress even after short-term RF exposure.


Assuntos
Células/metabolismo , Células/efeitos da radiação , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos da radiação , Células/citologia , Fluoresceínas/química , Glutationa/metabolismo , Humanos , Oxirredução , Estresse Oxidativo/efeitos da radiação , Ondas de Rádio , Espécies Reativas de Oxigênio/metabolismo
17.
J Biophotonics ; 11(1)2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-28671765

RESUMO

Actin, cytoskeleton protein forming microfilaments, play a crucial role in cellular motility. Here we show that exposure to very low levels of polarized light guide their orientation in-vivo within the live cell. Using a simple model to describe the role of actin-filament orientation in directional cellular motion, we demonstrate that the actin polymerization/depolymerization mechanism develops primarily along this direction and, under certain conditions, can lead to guidance of the cell movement. Our results also show a dose dependent increase in actin activity in direct correspondence to the level of laser irradiance. We found that total expression of Tau protein, which stabilize microtubules, was decreased by the irradiance, indicating that exposure to the light may change the activity of kinase, leading to increased cell activity.


Assuntos
Células/citologia , Células/efeitos da radiação , Luz , Movimento Celular/efeitos da radiação , Sobrevivência Celular/efeitos da radiação , Citoesqueleto/efeitos da radiação , Relação Dose-Resposta à Radiação , Células HEK293 , Humanos
18.
J Nucl Med Technol ; 45(4): 259-264, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29203550

RESUMO

The purpose of this paper is to review basic radiation biology and associated terminology to impart a better understanding of the importance of basic concepts of ionizing radiation interactions with living tissue. As health care workers in a field that utilizes ionizing radiation, nuclear medicine technologists are concerned about the possible acute and chronic effects of occupational radiation exposure. Technologists should have a clear understanding of what they are exposed to and how their safety could be affected. Furthermore, technologists should be knowledgeable about radiation effects so that they can adequately assuage possible patient fears about undergoing a nuclear medicine procedure. After reading this article, the nuclear medicine technologist will be familiar with basic radiation biology concepts; types of interactions of radiation with living tissue, and possible effects from that exposure; theoretic dose-response curves and how they are used in radiation biology; stochastic versus nonstochastic effects of radiation exposure, and what these terms mean in relation to both high- and low-dose radiation exposure; and possible acute and chronic radiation exposure effects.


Assuntos
Radiobiologia/métodos , Terminologia como Assunto , Células/efeitos da radiação , Relação Dose-Resposta à Radiação , Humanos , Exposição à Radiação/efeitos adversos
19.
Sci Rep ; 7: 40959, 2017 01 18.
Artigo em Inglês | MEDLINE | ID: mdl-28098222

RESUMO

We evaluated the dose-dependency and reversibility of radiation-induced injury in cardiac explant-derived cells (CDCs), a mixed cell population grown from heart tissues. Adult C57BL/6 mice were exposed to 0, 10, 50 and 250 mGy γ-rays for 7 days and atrial tissues were collected for experiments 24 hours after last exposure. The number of CDCs was significantly decreased by daily exposure to over 250 mGy. Interestingly, daily exposure to over 50 mGy significantly decreased the c-kit expression and telomerase activity, increased 53BP1 foci in the nuclei of CDCs. However, CD90 expression and growth factors production in CDCs were not significantly changed even after daily exposure to 250 mGy. We further evaluated the reversibility of radiation-induced injury in CDCs at 1 week and 3 weeks after a single exposure to 3 Gy γ-rays. The number and growth factors production of CDCs were soon recovered at 1 week. However, the increased expression of CD90 were retained at 1 week, but recovered at 3 weeks. Moreover, the decreased expression of c-kit, impaired telomerase activity, and increased 53BP1 foci were poorly recovered even at 3 weeks. These data may help us to find the most sensitive and reliable bio-parameter(s) for evaluating radiation-induced injury in CDCs.


Assuntos
Células/efeitos da radiação , Coração/efeitos da radiação , Miocárdio/citologia , Doses de Radiação , Lesões por Radiação/patologia , Animais , Sobrevivência Celular/efeitos da radiação , Ensaio de Imunoadsorção Enzimática , Perfilação da Expressão Gênica , Camundongos Endogâmicos C57BL
20.
Microsc Res Tech ; 80(5): 444-455, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28094894

RESUMO

Several new features of mitochondrial nucleoid and its surroundings in mammalian cells were described previously (Prachar, 2016). Very small details were observed using the improved transmission electron microscopy method, as described in the article. In the meantime, the method has again been improved to 2 Å resolutions in the cell section. The method described in detail in the present work is documented on the same records that were published in lower resolution in the work Prachar (2016), enabling comparison of the achieved resolution with the previous one. New records are also presented, showing extremely high resolution and thus implying the importance of the method. Potential use of this method in different fields is suggested.


Assuntos
Células/ultraestrutura , Microscopia Eletrônica de Transmissão/métodos , Exposição à Radiação/prevenção & controle , Animais , Linhagem Celular Tumoral , Células/efeitos da radiação , Elétrons , Retículo Endoplasmático/efeitos da radiação , Retículo Endoplasmático/ultraestrutura , Resinas Epóxi , Técnicas Histológicas , Leucemia L1210 , Camundongos , Mitocôndrias/efeitos da radiação , Mitocôndrias/ultraestrutura , Inclusão em Plástico/normas , Ribossomos/efeitos da radiação , Ribossomos/ultraestrutura , Fatores de Tempo
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