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spectrofluorometer, fluorescence, luminescence spectrometer, fluorescence spectrophotometer
This model was found at
882 locations
The model is used in
53 countries
Usage per year (up to 2020)
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144 related research fields
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About the Varian Cary Eclipse

The model Varian Cary Eclipse was found in 882 unique locations in 53 countries where it was mentioned from 2003 until recentlyIt is used by scientists in various research fields such as Biochemistry, Molecular Biology, Organic Chemistry, General Chemistry, and Physical and Theoretical Chemistry. The model is also used in General Medicine, Molecular Medicine, General Materials Science, Analytical Chemistry, Pharmaceutical Science, Drug Discovery, Spectroscopy, General Physics and Astronomy, Catalysis, General Biochemistry, Genetics and Molecular Biology, Inorganic Chemistry, Cell Biology, Bioengineering, Biophysics, Biotechnology, General Chemical Engineering, Chemistry, Pharmacology, Structural Biology, Computer Science Applications, Condensed Matter Physics, Microbiology, Biomedical Engineering, Biomaterials, and Surfaces and Interfaces.
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Research that uses the Varian Cary Eclipse

Miranda J. Nicholes, Christopher Williamson, Martyn Tranter, Alexandra Holland, Marian Yallop, Alexandre Anesio, 2020
Abstract. The surface (supraglacial) environment of the Greenland Ice Sheet (GrIS) is an active site for the storage, transformation and transport of carbon, which is driven by extremely high levels of solar radiation throughout the ablation season. Within the south west of the GrIS, blooms of Streptophyte micro-algae (hereafter glacier algae) at abundances of ~ 105 cell mL−1 dominate primary production in the surface ice and provide dissolved organic matter (DOM) to the heterotrophic bacterial community. Glacier algae contain photoprotective secondary phenolic pigment that comprises a large proportion of the cell (~ 4 % of the dry weight) and could represent a substantial, additional carbon source for the heterotrophic community. The transformation and degradation of DOM by solar radiation (photodegradation) and heterotrophic communities (biodegradation) represent two crucial controls on DOM composition and quantity; however, the influence of these processes within the surface ice is yet to be constrained. This study therefore assessed responses in the composition and quantity of two carbon sources (glacier algae secondary pigment and surface ice DOM) following exposure to UV, PAR, UV+PAR (photodegradation) and subsequent incubation with bacterial communities isolated from the ambient environment (biodegradation). Our results indicate that exposure to predominantly UV radiation altered the composition of glacier algal pigment and surface ice DOM; however, the quantity of DOM remained constant. Biodegradation caused the greatest changes to both DOM composition and quantity, particularly in surface ice DOM. Secondary pigment extracted from glacier algae was not a highly bioavailable source of carbon and did not support significant growth of surface ice heterotrophic bacterial communities. Conversely, low molecular weight compounds in surface ice DOM were rapidly utilised by heterotrophic bacteria supporting between a 3 and 9-fold increase in bacterial abundance over a 30-day incubation. We found that photodegradation of glacier algal pigment and surface ice DOM did not influence heterotrophic consumption. Photodegradation and biodegradation of DOM in the surface ice habitat are likely intimately linked and act as fundamental controls on the composition and quantity of DOM exported to downstream environments.
Byoung Chan Kim, Ji Young Choi, Kyung Kwang Joo, Seon Yeong Park, Ye Sung Song, Hee Jin Woo, Advances in High Energy Physics, 2021, 1-10, 2021
This paper proposes a new base material, a mixture of alcohol and water, for liquid scintillators. A possibility of using alcohol as a new detection solution in a particle detector is described. A liquid scintillator is widely used in various fields because of its high light yield. In addition, it is very important to develop a stable liquid scintillator for particle detectors or other medical applications. To date, there have been no previous R&D studies elsewhere for the use of alcohol in particle detectors, and no market products are available of this type. Thus, there is a room for improvement. This paper describes the brief synthesizing process of the alcohol-based liquid scintillator by varying the mixing ratio of each component that makes up the liquid scintillator. The several feasible physical and optical properties of an alcohol-based liquid scintillator were investigated and presented. Finally, as one of its applications, a range (beam-path length) measurement using an electron beam in medical physics is introduced after irradiating an alcohol-based liquid scintillator with electron beam energies of 6~12 MeV. The measurement results were compared with a Monte Carlo simulation, Novalis Tx, a phantom, and a CT image. In the near future, the new alcohol-based liquid scintillator could be used for particle detector or medical imaging applications.
Antonina Naumenko, Nataliya Kutsevol, International Journal of Polymer Science, 2021, 1-7, 2021
Chlorin e6 and its derivatives are the basis of a number of drugs used in medicine in the treatment of various diseases, including cancer, by photodynamic therapy. Nonpolar derivatives of Chlorin e6—dimethyl ether of Chlorin e6 (DME Ce6) and trimethyl ether of Chlorin e6 (TME Ce6)—are actively studied for application during photodynamic therapy. In this work, based on the electron optical absorption spectra, the interaction of photosensitizer molecules with branched star-like copolymer dextran-graft-polyacrylamide in anionic form was investigated and the possibility of using the latter as a carrier for drug delivery to tumor cells was suggested.
Matthias Bauer, Jakob Steube, Ayla Päpcke, Olga Bokareva, Thomas Reuter, Serhiy Demeshko, Roland Schoch, Stephan Hohloch, Franc Meyer, Katja Heinze, Oliver Kühn, Stefan Lochbrunner, 2020
Abstract Photoactive compounds are essential for photocatalytic and luminescent applications, such as photoredox catalysis or light emitting diodes. However, the substitution of noble metals, which are almost exclusively used, by base metals remains a major challenge on the way to a more sustainable world.1 Iron is a dream candidate for this ambitious aim.2 But compared to noble metal complexes that show long-lived metal-to-ligand charge-transfer (MLCT) states, realization of emissive and photoactive iron complexes is demanding, due to the fast deactivation of charge transfer states into non-emissive inactive states. No MLCT emission has been observed for monometallic iron complexes before. Consequently, dual emission could also not yet be realized with iron complexes, as it is a very rare property even of noble metal compounds. Here we report the FeIII complex [Fe(ImP)2][PF6] (HImP = 1,1’-(1,3-phenylene)bis(3-methyl-1-imidazol-2-ylidene)), showing Janus-type dual emission by combining LMCT (ligand-to-metal charge transfer) with MLCT luminescence. The respective excited states are characterized by a record lifetime of τMLCT = 4.2 ns, and a moderate τLMCT = 0.2 ns. Only two emissive FeIII compounds are known so far and they show LMCT luminescence only.3,4 The unique properties of the presented complex are caused by the specific ligand design combining four N-heterocyclic carbenes with two cyclometalating groups, using the σ-donor strength of six carbon atoms and the acceptor capabilities of the central phenyl rings. Spectroscopically, doublet manifolds could be identified in the deactivation process, while (TD)DFT analysis revealed the presence of quartets as well. With three key advancements of realizing the first iron complex showing dual luminescence, a MLCT luminescence and a world record MLCT lifetime, the results constitute a basis for future application of iron complexes as white light emitters and new photocatalytic reactions making use of the Janus-type properties of the developed complex.
Margarida Rodrigues, Payel Bhattacharjee, Ann Brinkmalm, Dung Do, Colin Pearson, Suman De, Aleks Ponjavic, Juan Varela, Francesco Ruggeri, Isabelle Baudrexel, Ji Lee, Alexander Carr, Klara Kulenkampff, Tuomas Knowles, Henrik Zetterberg, Thomas Snaddon, Sonia Gandhi, Steven Lee, David Klenerman, 2020
Abstract The composition of soluble toxic protein aggregates formed in vivo is currently unknown in neurodegenerative diseases, due to their ultra-low concentration in human biofluids and their high degree of heterogeneity. We introduce the structure-specific chemical antibody; a Y shaped, bioinspired small molecule with a dimeric region to mimic avidity, and an attachment region to mimic the Fc region. Our probe, capture molecule for amyloid precipitation (CAP-1), consists of a derivative of Pittsburgh compound B (dimer) to target the cross β-sheets of amyloids and a biotin moiety for surface immobilization. By coupling CAP-1 to magnetic beads, we targeted the amyloid structure of protein aggregates in human cerebrospinal fluid, isolated them for analysis and then characterised them using single-molecule fluorescence imaging and mass spectrometry. AP allows unbiased determination of the molecular composition and structural features of the in vivo aggregates, formed in neurodegenerative diseases, that are present in biofluids.
Xiang Xia, Ran Wang, Yingqi Hu, WeiJian Liu, Ting Liu, Wen Sun, Jiangli Fan, Xiaojun Peng, Frontiers in Chemistry, 9, 2021
Lipid droplets (LDs), an extremely important cellular organelle, are responsible for the storage of neutral lipids in multiple biological processes, which could be a potential target site for photodynamic therapy (PDT) of cancer. Herein, a lipid droplet–targeted photosensitizer (BODSeI) is developed, allowing for fluorescence imaging–guided PDT. Owing to the location of lipid droplets, BODSeI demonstrates enhanced PDT efficiency with an extremely low IC50 value (around 125 nM). Besides, BODSeI shows good biocompatibility and high photostability. Therefore, BODSeI is promising for droplet-location PDT, which may trigger wide interest for exploring the pathway of lipid droplet–location PDT.
Rathi Saravanan, Yeu Khai Choong, Chun Hwee Lim, Li Ming Lim, Jitka Petrlova, Artur Schmidtchen, Frontiers in Immunology, 12, 2021
Cell-free DNA (cfDNA) is the major structural component of neutrophil extracellular traps (NETs), an innate immune response to infection. Antimicrobial proteins and peptides bound to cfDNA play a critical role in the bactericidal property of NETs. Recent studies have shown that NETs have procoagulant activity, wherein cfDNA triggers thrombin generation through activation of the intrinsic pathway of coagulation. We have recently shown that thrombin binds to NETs in vitro and consequently can alter the proteome of NETs. However, the effect of NETs on thrombin is still unknown. In this study, we report that DNA binding leads to thrombin autolysis and generation of multiple thrombin-derived C-terminal peptides (TCPs) in vitro. Employing a 25-residue prototypic TCP, GKY25 (GKYGFYTHVFRLKKWIQKVIDQFGE), we show that TCPs bind NETs, thus conferring mutual protection against nuclease and protease degradation. Together, our results demonstrate the complex interplay between coagulation, NET formation, and thrombin cleavage and identify a previously undisclosed mechanism for formation of TCPs.
Lisa Marx, Enrico F. Semeraro, Johannes Mandl, Johannes Kremser, Moritz P. Frewein, Nermina Malanovic, Karl Lohner, Georg Pabst, Frontiers in Medical Technology, 3, 2021
We coupled the antimicrobial activity of two well-studied lactoferricin derivatives, LF11-215 and LF11-324, in Escherichia coli and different lipid-only mimics of its cytoplasmic membrane using a common thermodynamic framework for peptide partitioning. In particular, we combined an improved analysis of microdilution assays with ζ-potential measurements, which allowed us to discriminate between the maximum number of surface-adsorbed peptides and peptides fully partitioned into the bacteria. At the same time, we measured the partitioning of the peptides into vesicles composed of phosphatidylethanolamine (PE), phosphatidylgylcerol (PG), and cardiolipin (CL) mixtures using tryptophan fluorescence and determined their membrane activity using a dye leakage assay and small-angle X-ray scattering. We found that the vast majority of LF11-215 and LF11-324 readily enter inner bacterial compartments, whereas only 1−5% remain surface bound. We observed comparable membrane binding of both peptides in membrane mimics containing PE and different molar ratios of PG and CL. The peptides' activity caused a concentration-dependent dye leakage in all studied membrane mimics; however, it also led to the formation of large aggregates, part of which contained collapsed multibilayers with sandwiched peptides in the interstitial space between membranes. This effect was least pronounced in pure PG vesicles, requiring also the highest peptide concentration to induce membrane permeabilization. In PE-containing systems, we additionally observed an effective shielding of the fluorescent dyes from leakage even at highest peptide concentrations, suggesting a coupling of the peptide activity to vesicle fusion, being mediated by the intrinsic lipid curvatures of PE and CL. Our results thus show that LF11-215 and LF11-324 effectively target inner bacterial components, while the stored elastic stress makes membranes more vulnerable to peptide translocation.
Margarita Shleeva, Alexander Savitsky, Arseny Kaprelyants, Frontiers in Microbiology, 11, 2020
Pathogenic non-spore forming bacteria enter a dormant state under stressful conditions, which likely allows them to acquire resistance to various antibiotics. This work revealed the efficient formation of dormant “non-culturable” (NC) Corynebacterium jeikeium cells in stationary phase upon gradual acidification of the growth medium. Such cells were unable to form colonies and existed in a prolonged stationary phase. At an early stage of dormancy (approximately 14 days post-inoculation), dormant cells are able for resuscitation in liquid medium. However, those stored for long time in dormant state needed addition of supernatant taking from active C. jeikeium cultures for successful resuscitation. NC cells possessed low RNA synthesis and significant tolerance to antibiotics (rifampicin and vancomycin). They also accumulated free porphyrins, and 5-aminolevulinic acid addition enhanced free porphyrin accumulation which makes them potentially sensitive to photodynamic inactivation (PDI). PDI of dormant bacteria was accomplished by exposing cells to a 565 nm wavelength of light using a SOLIS-4C light-emitting diode for 60 min. This revealed that increased porphyrin concentrations were correlated with elevated PDI sensitivity. Results shown here demonstrate the potential utility of employing PDI to minimize levels of dormant, persistent corynebacteria and the C. jeikeium dormancy model developed here may be useful for finding new drugs and techniques for combatting persistent corynebacteria.
Priyank Singhvi, Juhi Verma, Neha Panwar, Tabiya Qayoom Wani, Akansha Singh, Md. Qudratullah, Arnab Chakraborty, Ankit Saneja, Debi P. Sarkar, Amulya K. Panda, Frontiers in Microbiology, 12, 2021
Understanding the structure–function of inclusion bodies (IBs) in the last two decades has led to the development of several mild solubilization buffers for the improved recovery of bioactive proteins. The recently developed freeze–thaw-based inclusion body protein solubilization method has received a great deal of attention due to its simplicity and cost-effectiveness. The present report investigates the reproducibility, efficiency, and plausible mechanism of the freeze–thaw-based IB solubilization. The percentage recovery of functionally active protein species of human growth hormone (hGH) and L-asparaginase from their IBs in Escherichia coli and the quality attributes associated with the freeze–thaw-based solubilization method were analyzed in detail. The overall yield of the purified hGH and L-asparaginase protein was found to be around 14 and 25%, respectively. Both purified proteins had functionally active species lower than that observed with commercial proteins. Biophysical and biochemical analyses revealed that the formation of soluble aggregates was a major limitation in the case of tough IB protein like hGH. On the other hand, the destabilization of soft IB protein like L-asparaginase led to the poor recovery of functionally active protein species. Our study provides insight into the advantages, disadvantages, and molecular–structural information associated with the freeze–thaw-based solubilization method.
Renata Tisi, Michela Spinelli, Alessandro Palmioli, Cristina Airoldi, Paolo Cazzaniga, Daniela Besozzi, Marco S. Nobile, Elisa Mazzoleni, Simone Arnhold, Luca De Gioia, Rita Grandori, Francesco Peri, Marco Vanoni, Elena Sacco, Frontiers in Molecular Biosciences, 8, 2021
Ras oncoproteins play a crucial role in the onset, maintenance, and progression of the most common and deadly human cancers. Despite extensive research efforts, only a few mutant-specific Ras inhibitors have been reported. We show that cmp4–previously identified as a water-soluble Ras inhibitor– targets multiple steps in the activation and downstream signaling of different Ras mutants and isoforms. Binding of this pan-Ras inhibitor to an extended Switch II pocket on HRas and KRas proteins induces a conformational change that down-regulates intrinsic and GEF-mediated nucleotide dissociation and exchange and effector binding. A mathematical model of the Ras activation cycle predicts that the inhibitor severely reduces the proliferation of different Ras-driven cancer cells, effectively cooperating with Cetuximab to reduce proliferation even of Cetuximab-resistant cancer cell lines. Experimental data confirm the model prediction, indicating that the pan-Ras inhibitor is an appropriate candidate for medicinal chemistry efforts tailored at improving its currently unsatisfactory affinity.
Sathish Kumar, Akshay Kapadia, Sandra Theil, Pranav Joshi, Florian Riffel, Michael T. Heneka, Jochen Walter, Frontiers in Molecular Neuroscience, 13, 2021
Aggregation and deposition of amyloid-β (Aβ) peptides in extracellular plaques and in the cerebral vasculature are prominent neuropathological features of Alzheimer’s disease (AD) and closely associated with the pathogenesis of AD. Amyloid plaques in the brains of most AD patients and transgenic mouse models exhibit heterogeneity in the composition of Aβ deposits, due to the occurrence of elongated, truncated, and post-translationally modified Aβ peptides. Importantly, changes in the deposition of these different Aβ variants are associated with the clinical disease progression and considered to mark sequential phases of plaque and cerebral amyloid angiopathy (CAA) maturation at distinct stages of AD. We recently showed that Aβ phosphorylated at serine residue 26 (pSer26Aβ) has peculiar characteristics in aggregation, deposition, and neurotoxicity. In the current study, we developed and thoroughly validated novel monoclonal and polyclonal antibodies that recognize Aβ depending on the phosphorylation-state of Ser26. Our results demonstrate that selected phosphorylation state-specific antibodies were able to recognize Ser26 phosphorylated and non-phosphorylated Aβ with high specificity in enzyme-linked immunosorbent assay (ELISA) and Western Blotting (WB) assays. Furthermore, immunofluorescence analyses with these antibodies demonstrated the occurrence of pSer26Aβ in transgenic mouse brains that show differential deposition as compared to non-phosphorylated Aβ (npAβ) or other modified Aβ species. Notably, pSer26Aβ species were faintly detected in extracellular Aβ plaques but most prominently found intraneuronally and in cerebral blood vessels. In conclusion, we developed new antibodies to specifically differentiate Aβ peptides depending on the phosphorylation state of Ser26, which are applicable in ELISA, WB, and immunofluorescence staining of mouse brain tissues. These site- and phosphorylation state-specific Aβ antibodies represent novel tools to examine phosphorylated Aβ species to further understand and dissect the complexity in the age-related and spatio-temporal deposition of different Aβ variants in transgenic mouse models and human AD brains.
Zhiwei Zhang, Xiaowei Zhang, Lei Meng, Mengqi Gong, Jian Li, Wen Shi, Jiuchun Qiu, Yajuan Yang, Jianping Zhao, Ya Suo, Xue Liang, Xinghua Wang, Gary Tse, Ning Jiang, Guangping Li, Yungang Zhao, Tong Liu, Frontiers in Pharmacology, 12, 2021
Background: Oxidative stress contributes to adverse atrial remodeling in diabetes mellitus. This remodeling can be prevented by the PPAR-γ agonist pioglitazone via its antioxidant and anti-inflammatory effects. In this study, we examined the molecular mechanisms underlying the protective effects of pioglitazone on atrial remodeling in a rabbit model of diabetes.Methods: Rabbits were randomly divided into control, diabetic, and pioglitazone-treated diabetic groups. Echocardiographic, hemodynamic, and electrophysiological parameters were measured. Serum PPAR-γ levels, serum and tissue oxidative stress and inflammatory markers, mitochondrial morphology, reactive oxygen species (ROS) production rate, respiratory function, and mitochondrial membrane potential (MMP) levels were measured. Protein expression of the pro-fibrotic marker TGF-β1, the PPAR-γ coactivator-1α (PGC-1α), and the mitochondrial proteins (biogenesis-, fusion-, and fission-related proteins) was measured. HL-1 cells were transfected with PGC-1α small interfering RNA (siRNA) to determine the underlying mechanisms of pioglitazone improvement of mitochondrial function under oxidative stress.Results: The diabetic group demonstrated a larger left atrial diameter and fibrosis area than the controls, which were associated with a higher incidence of inducible atrial fibrillation (AF). The lower serum PPAR-γ level was associated with lower PGC-1α and higher NF-κB and TGF-β1 expression. Lower mitochondrial biogenesis (PGC-1α, NRF1, and TFAM)-, fusion (Opa1 and Mfn1)-, and fission (Drp1)-related proteins were detected. Mitochondrial swelling, higher mitochondrial ROS, lower respiratory control rate, and lower MMP were observed. The pioglitazone group showed a reversal of structural remodeling and a lower incidence of inducible AF, which were associated with higher PPAR-γ and PGC-1α. The pioglitazone group had lower NF-κB and TGF-β1 expression levels, whereas biogenesis-, fusion-, and fission-related protein expression was higher. Further, mitochondrial structure and function were improved. In HL-1 cells, PGC-1α siRNA transfection blunted the effect of pioglitazone on Mn-SOD protein expression and MMP collapse in H2O2-treated cells.Conclusion: Diabetes mellitus induces adverse atrial structural, electrophysiological remodeling, and mitochondrial damage and dysfunction. Pioglitazone prevented these abnormalities through the PPAR-γ/PGC-1α pathway.
Renhua Sun, Chengjun Song, Yuanze Sun, Qian Wang, Hongbin Liu, Jie Wang, Agronomy, 10, 1316 (9), 2020
Despite the essential roles of soil dissolved organic matter (DOM) and soil microbes in agro-ecosystems, we still have a limited understanding of the extent by which they are impacted by agronomic strategies in ecological and conventional farming. Using three-dimensional fluorescence excitation–emission matrices (3D-EEM) and high-throughput microbial sequencing, the characteristics of soil DOM and microbiota under realistic field conditions were estimated in the farming soils with long-term ecological (EM) and conventional management (CM). Specifically, the role of hedgerows in the ecologically managed land (EMH) was assessed. The total fluorescent intensity of soil DOM in the EMH system was significantly higher than the values in CM and EM systems. Additionally, the five normalized excitation–emission area volumes from regional integration analysis increased in the order CM < EM < EMH. In comparison with CM and EM soils, the hedgerow significantly increased the evenness of the bacterial communities in the EMH system, whereas no differences were found for the alpha-diversity of eukaryotic communities. The composition of soil microbiota was significantly distinct among the three farming systems, with a hedgerow-specific effect on bacterial community and a management-specific effect on eukarya. The predicted functional profiles indicated that the hedgerow showed a higher contribution to the dissimilarity of bacterial functions. Furthermore, the distinction of the soil microbiota was modulated by the soil DOM composition and significantly positive correlations between the microbiota involved in nutrient cycling and soil DOM were observed. The findings in this work strengthen our understanding of the different responses of bacterial and eukaryotic communities under the long-term ecological management and highlight the beneficial roles of hedgerows in increasing organic matter and modulating community assembly.
Ana Belén Uceda, Josefa Donoso, Juan Frau, Bartolomé Vilanova, Miquel Adrover, Antioxidants, 10, 315 (2), 2021
Frataxin is a mitochondrial protein which deficiency causes Friedreich’s ataxia, a cardio-neurodegenerative disease. The lack of frataxin induces the dysregulation of mitochondrial iron homeostasis and oxidative stress, which finally causes the neuronal death. The mechanism through which frataxin regulates the oxidative stress balance is rather complex and poorly understood. While the absence of human (Hfra) and yeast (Yfh1) frataxins turn out cells sensitive to oxidative stress, this does not occur when the frataxin gene is knocked-out in E. coli. To better understand the biological roles of Hfra and Yfh1 as endogenous antioxidants, we have studied their ability to inhibit the formation of reactive oxygen species (ROS) from Cu2+- and Fe3+-catalyzed degradation of ascorbic acid. Both proteins drastically reduce the formation of ROS, and during this process they are not oxidized. In addition, we have also demonstrated that merely the presence of Yfh1 or Hfra is enough to protect a highly oxidation-prone protein such as α-synuclein. This unspecific intervention (without a direct binding) suggests that frataxins could act as a shield to prevent the oxidation of a broad set of intracellular proteins, and reinforces that idea that frataxin can be used to prevent neurological pathologies linked to an enhanced oxidative stress.
Megan J. Reiniers, Lianne R. de Haan, Laurens F. Reeskamp, Mans Broekgaarden, Rowan F. van Golen, Michal Heger, Antioxidants, 10, 674 (5), 2021
Numerous liver pathologies encompass oxidative stress as molecular basis of disease. The use of 2′,7′-dichlorodihydrofluorescein-diacetate (DCFH2-DA) as fluorogenic redox probe is problematic in liver cell lines because of membrane transport proteins that interfere with probe kinetics, among other reasons. The properties of DCFH2-DA were analyzed in hepatocytes (HepG2, HepaRG) to characterize methodological issues that could hamper data interpretation and falsely skew conclusions. Experiments were focused on probe stability in relevant media, cellular probe uptake/retention/excretion, and basal oxidant formation and metabolism. DCFH2-DA was used under optimized experimental conditions to intravitally visualize and quantify oxidative stress in real-time in HepG2 cells subjected to anoxia/reoxygenation. The most important findings were that: (1) the non-fluorescent DCFH2-DA and the fluorescent DCF are rapidly taken up by hepatocytes, (2) DCF is poorly retained in hepatocytes, and (3) DCFH2 oxidation kinetics are cell type-specific. Furthermore, (4) DCF fluorescence intensity was pH-dependent at pH < 7 and (5) the stability of DCFH2-DA in cell culture medium relied on medium composition. The use of DCFH2-DA to measure oxidative stress in cultured hepatocytes comes with methodological and technical challenges, which were characterized and solved. Optimized in vitro and intravital imaging protocols were formulated to help researchers conduct proper experiments and draw robust conclusions.
Bo Shao, Li Mao, Miao Tang, Zhu-Ying Yan, Jie Shao, Chun-Hua Huang, Zhi-Guo Sheng, Ben-Zhan Zhu, Antioxidants, 10, 798 (5), 2021
Caffeic acid phenethyl ester (CAPE) and its structurally-related caffeic acid (CA), ferulic acid (FA) and ethyl ferulate (EF) are constituents of honeybee propolis that have important pharmacological activities. This study found that CAPE—but not CA, FA, and EF—could effectively prevent cellular DNA damage induced by overloaded iron through decreasing the labile iron pool (LIP) levels in HeLa cells. Interestingly, CAPE was found to be more effective than CA in protecting against plasmid DNA damage induced by Fe(II)–H2O2 or Fe(III)–citrate–ascorbate-H2O2 via the inhibition of hydroxyl radical (•OH) production. We further provided more direct and unequivocal experimental evidences for the formation of inactive CAPE/CA–iron complexes. CAPE was found to have a stronger iron-binding ability and a much higher lipophilicity than CA. Taken together, we propose that the esterification of the carboxylic moiety with phenethyl significantly enhanced the iron-binding ability and lipophilicity of CAPE, which is also responsible for its potent protection against iron-mediated cellular DNA damage. A study on the iron coordination mechanism of such natural polyphenol antioxidants will help to design more effective antioxidants for the treatment and prevention of diseases caused by metal-induced oxidative stress, as well as help to understand the structure–activity relationships of these compounds.
Lidija Milkovic, Tea Vukovic, Neven Zarkovic, Franz Tatzber, Egils Bisenieks, Zenta Kalme, Imanta Bruvere, Zaiga Ogle, Janis Poikans, Astrida Velena, Gunars Duburs, Antioxidants, 7, 123 (9), 2018
Oxidative stress has been implicated in pathophysiology of different human stress- and age-associated disorders, including osteoporosis for which antioxidants could be considered as therapeutic remedies as was suggested recently. The 1,4-dihydropyridine (DHP) derivatives are known for their pleiotropic activity, with some also acting as antioxidants. To find compounds with potential antioxidative activity, a group of 27 structurally diverse DHPs, as well as one pyridine compound, were studied. A group of 11 DHPs with 10-fold higher antioxidative potential than of uric acid, were further tested in cell model of human osteoblast-like cells. Short-term combined effects of DHPs and 50 µM H2O2 (1-h each), revealed better antioxidative potential of DHPs if administered before a stressor. Indirect 24-h effect of DHPs was evaluated in cells further exposed to mild oxidative stress conditions induced either by H2O2 or tert-butyl hydroperoxide (both 50 µM). Cell growth (viability and proliferation), generation of ROS and intracellular glutathione concentration were evaluated. The promotion of cell growth was highly dependent on the concentrations of DHPs used, type of stressor applied and treatment set-up. Thiocarbatone III-1, E2-134-1 III-4, Carbatone II-1, AV-153 IV-1, and Diethone I could be considered as therapeutic agents for osteoporosis although further research is needed to elucidate their bioactivity mechanisms, in particular in respect to signaling pathways involving 4-hydroxynoneal and related second messengers of free radicals.
Marianna Lauricella, Valentina Lo Galbo, Cesare Cernigliaro, Antonella Maggio, Antonio Palumbo Piccionello, Giuseppe Calvaruso, Daniela Carlisi, Sonia Emanuele, Michela Giuliano, Antonella D’Anneo, Antioxidants, 8, 422 (10), 2019
Ethanolic extracts from Mangifera indica L. have been proved to possess anti-tumor properties in many cancer systems. However, although most effects have been demonstrated with fruit pulp extract, the underlying molecular mechanisms of mango peel are still unclear. This study was designed to explore the effects of mango peel extract (MPE) on colon cancer cell lines. MPE affected cell viability and inhibited the colony formation trend of tumor cells, while no effects were observed in human dermal fibroblasts used as a non-cancerous cell line model. These events were a consequence of the induction of apoptosis associated to reactive oxygen species (ROS) production, activation of players of the oxidative response such as JNK and ERK1/2, and the increase in Nrf2 and manganese superoxide dismutase (MnSOD). Significantly, mango peel-activated stress triggered a DNA damage response evidenced by the precocious phosphorylation of histone 2AX (γH2AX), as well as phosphorylated Ataxia telangiectasia-mutated (ATM) kinase and p53 upregulation. Mango peel extract was also characterized, and HPLC/MS (High Performance Liquid Chromatography/Mass Spectrometry) analysis unveiled the presence of some phenolic compounds that could be responsible for the anti-cancer effects. Collectively, these findings point out the importance of the genotoxic stress signaling pathway mediated by γH2AX in targeting colon tumor cells to apoptosis.
undefined Khrabrova, undefined Loiko, undefined Tolkacheva, undefined Cherepanova, undefined Zvereva, undefined Kirsanova, undefined Gromova, Biomolecules, 10, 8 (1), 2019
In mammals, DNA methylation is necessary for the maintenance of genomic stability, gene expression regulation, and other processes. During malignant diseases progression, changes in both DNA methylation patterns and DNA methyltransferase (MTase) genes are observed. Human de novo MTase DNMT3A is most frequently mutated in acute myeloid leukemia (AML) with a striking prevalence of R882H mutation, which has been extensively studied. Here, we investigate the functional role of the missense mutations (S714C, R635W, R736H, R771L, P777R, and F752V) found in the catalytic domain of DNMT3A in AML patients. These were accordingly mutated in the murine Dnmt3a catalytic domain (S124C, R45W, R146H, R181L, P187R, and F162V) and in addition, one-site CpG-containing DNA substrates were used as a model system. The 3–15-fold decrease (S124C and P187R) or complete loss (F162V, R45W, and R146H) of Dnmt3a-CD methylation activity was observed. Remarkably, Pro 187 and Arg 146 are not located at or near the Dnmt3a functional motives. Regulatory protein Dnmt3L did not enhance the methylation activity of R45W, R146H, P187R, and F162V mutants. The key steps of the Dnmt3a-mediated methylation mechanism, including DNA binding and transient covalent intermediate formation, were examined. There was a complete loss of DNA-binding affinity for R45W located in the AdoMet binding region and for R146H. Dnmt3a mutants studied in vitro suggest functional impairment of DNMT3A during pathogenesis.
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