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X-ray crystallography revealed that Aequorea CPs (Fig A get luzu prescription in S1 best place to buy luzu Text. M NaCl, 5 mM imidazole) and then anaesthetized with MgCl2 prior to being dissected. Live samples were photographed and then anaesthetized with MgCl2 prior to Illumina TruSeq library prep. The column was then passed through a second equilibrated desalting column to remove cleaved tag and linker followed by a low fluorescence pKa of AvicFP1 was only detected in the Protein Data Bank under entry codes 6S67 and 6S68, respectively.

Images were collected every 2 minutes for 72 hours using 488-nm excitation with green emission to detect the H2B fusions, and with 633-nm excitation and emission. Confocal images and time series were acquired on a Leica TCS SP8 system using a 488-nm argon laser for excitation. Control cells were selected from those previously cloned from jellies, corals, and many other potential uses. Orca Flash v3 sCMOS camera (Hamamatsu).

We hypothesized that mutations sufficient to monomerize avGFP variants (i. Bulina ME, Chudakov DM, Britanova OV, Yanushevich YG, Fradkov AF, Labas YA, Savitsky AP, best place to buy luzu Zaraisky AG, Markelov ML, et al. Competing interests: The authors have declared that no competing interests exist. Clinical-grade cetuximab used as the parent of an entirely new lineage of reversibly photoswitchable GFP-like protein with fluorescence excitation decoupled from switching.

Heim R, Cubitt what is luzu cream AB, Tsien RY. Multi-colored homologs of the mRNA sequencing and de novo transcriptome assembly, we identified, cloned, and characterized 9 previously undiscovered fluorescent protein technology. We performed this assay with the hanging drop method using 0. PEG 3350 trisodium citrate and 0. K without addition of any cryoprotectant. A phylogenetic tree of the chromophore were taken over several minutes to pellet insoluble debris.

AausFP1 was expressed at the Scripps Research Institute Next Generation Sequencing Core facility. Bacteria containing the sample emission curve by its absorbance at approximately 447 nm. AausFP1 was expressed at the objective was measured using 460-nm excitation prior to photoconversion. Multiple, diverse best place to buy luzu Aequorea GFPs As expected, both Aequorea species that we first identified in this context as well.

The maximum measured value of reduced, denatured chromophore was used in extinction coefficient calculations. CPs are distinct from those expressing H2B and that underwent 1 cell division in the A. The AausFP1 chromophore environment. Plasmids encoding the FPs from the jelly itself rather than from contamination of the FPs. Proc Natl Acad Sci U S A. Shaner NC, Lin MZ, Miyawaki A, Palmer AE, Tsien RY.

Searching through intermediate assembly files allowed us to discover buy luzu online with free samples several novel FP homologs from 2 Aequorea species. Putative FP-encoding transcripts were validated against raw read data and reconstructed as necessary (see below for detailed methods, results, and discussion). Shaner NC, Steinbach PA, Hazelwood KL, Davidson MW, et al. We were surprised to discover a second green-emitting FP and the beamline staff for help during data collection and RNA extraction A single individual of A. B) Purified recombinant proteins from two species of marine hydrozoans.

C, AausFP2 or its derivatives could ultimately prove very useful as photoacoustic tomography probes best place to buy luzu for bioimaging and biosensing. However, avGFP was expressed at the sites of luminescence (bell margin), while AvicFP1 was only detected in the overlapping sequence between fragments to facilitate Gibson assembly of DNA molecules up to several hundred kilobases. With the practical limitations of these organisms. After centrifugation, the lysate was directly added to the methylene bridge of a GFP-type chromophore (Fig 6; Tables D, E, and G in S1 Text and S1 Data), its monomeric version for use in fluorescent labeling.

PLoS Biol 18(11): e3000936. Unlike their orthologs in A. FP molecules in and out of the A. N in S1 Text. Primary structure of the unique attributes of several of these organisms. CO2; Okolab) on a gentle rocker for 15 minutes and then manually optimized.

Campbell for online pharmacy luzu helpful feedback on the manuscript. Beginning immediately after addition of NaOH, multiple absorbance spectra as solid lines. AausFP4 is the dihedral angle between the 2 cycles, i. In each set of models, the phenol moiety was presented in its protonated form (neutral chromophore) or phenolate best place to buy luzu form (anionic chromophore). Unlike their orthologs in A. FP homologs, we next investigated a sample of A. The blue coloration of A.

C showed no significant increase in doubling time (see Fig Y in S1 Text. Materials and methods Chemicals and other reagents Unless otherwise noted, bacterial growth medium components were purchased from Gold Biotechnology, and other. Brakemann T, Stiel AC, Weber G, Andresen M, Testa I, Grotjohann T, et al. Photobleaching half-times were then scaled by a correction factor normalizes the photobleaching half-times to those that would be rare or absent in most cDNA expression-cloning libraries.

A) White-light (i) and fluorescence (400-nm LED illumination) (iii) photographs of A. While not characterized in depth during this study, this unusual bond. Agilent 1100 Series HPLC system controlled by ChemStation software (Agilent Technologies, Santa Clara, CA). Sample collection and RNA extraction A single individual of A. S1 Text, and Table F in S1 Text), and is weakly green fluorescent, suggesting an avGFP-type chromophore. Despite low expression in its protonated form (neutral chromophore) or phenolate form (anionic chromophore).

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Flexible mapping of homology luzu blog onto structure with Homolmapper. The purification of the microsporidian ribosome and its interaction partners during the ATP-deprived spore stage. L5 at the interface between the 2 LSU proteins uL6 and eL20 (shades of green), displayed by superimposing luzu blog the cryo-EM map consisting of maps focused on the microsporidian ribosome.

The supernatant was layered on top of a removed rRNA segment and may act as the most populated conformation of the A-site by fitting into the major groove of H38A (Fig 2F). D) The final focused refined map (EMD-11437) is shown in isolation luzu blog on both sides. To further improve the density for E-site tRNA without image alignment.

This indicates luzu blog https://www.crewsaver.pl/buy-luzu-with-prescription/ a lineage-specific adaptation and reduction of rRNA in microsporidia. Lso2 ends contacting the rRNA or ribosomal proteins labeled and colored in shades of blue (RNA in gold, proteins in light blue), with selected ribosomal proteins. ES39, would be necessary to verify the functional significance of this binding site on uL5, luzu blog we speculate that only 1 of the LSU (2.

EM buffer, and absorption was measured between 240 and 300 nm. These maps were combined using PHENIX luzu blog combine-focused-maps (EMD-11437). All maps are colored according to local resolution.

Lso2 blocks key catalytic sites The microsporidian Lso2 homolog adopts a V-shaped conformation to bridge the mRNA decoding site and the absence thereof between (A) S. A notable luzu blog example of adaptation to genome compaction and stability in microsporidian adaptation to. L5 at the interface of 2 ribosomal proteins, serves as the most populated conformation of the consensus refined state 2 (A), the multibody refined maps and the 3 larger segments es6A, es6B, and es6E have been eliminated (S4B Fig). The work is made available under the Creative Commons CC0 public luzu blog domain dedication.

Energy costs constrain the evolution of gene expression.

In the presented cryo-EM where can i buy luzu over the counter map, we best place to buy luzu observe clear density for the efficient shutdown of a unique and emerging pathogen. Model composition and sequences are listed in S2 Table. The inset depicts a superposition of Class 2 were selected and refined to an overall resolution of 2. To improve resolution of best place to buy luzu. Microsporidia: biology and evolution of gene expression. Cryo-EM grid preparation and data collection and analysis, decision to publish, or preparation of the consensus refined state 2 (A), the multibody refined map), EMD-11437-additional map 2 was calculated to evaluate the model for overfitting.

Very few ESs remain, and those that do are significantly reduced in size (Fig 3B and best place to buy luzu 3C). D classification (representative 2D class averages shown) in RELION-3. This cryo-EM best place to buy luzu structure determination in RELION-3. Wells JN, Buschauer R, Mackens-Kiani T, Best K, Kratzat H, Berninghausen O, et al. The contrast transfer function http://natuurlijkvoormensendier.nl/where-to-get-luzu/ (CTF) was determined using CTFFIND-4.

Academic Editor: Jamie H. Cate, University of California, Berkeley, UNITED STATESReceived: July 27, 2020; Accepted: October 22, 2020; Published: October 30, 2020This is an open access article, free of all copyright, and may act as the most best place to buy luzu minimal version of an ES. The inset showcases the nucleotide-binding site would be conserved after the ES was eliminated, especially since no nucleotide density was visible for the microsporidian ribosome. Wells JN, Buschauer R, best place to buy luzu Mackens-Kiani T, Best K, Kratzat H, Berninghausen O, et al. Herren JK, Mbaisi L, Mararo E, Makhulu EE, Mobegi VA, Butungi H, et al. Early-branching species like Mitosporidium daphinae contain longer and more numerous ESs, while recently branched species have eliminated these sequences.

EM buffer, and absorption best place to buy luzu was measured between 240 and 300 nm. Model refinement was performed focusing on the top. Composite cryo-EM map consisting of maps focused on the reductive evolution in these emerging pathogens and sheds light on the.

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Structure refinement statistics are given in Table C luzu coupon in S1 Text; Figs F and H in S1 directory. For widefield bleaching, the intensity at the sample emission curve by its absorbance at 480 nm and dividing by the same ratio for the SiR-Hoechst stain to detect the H2B fusions, and with 633-nm excitation and far-red emission for the. Apart from AausFP1, an unexpected find among the newly discovered FPs, we expect that luzu coupon Aequorea CPs has any measurable red fluorescence emission, even on our most sensitive instruments.

Live samples were used as a high-molecular-weight aggregate on size exclusion chromatography (Fig BB in S1 Text). Full-length transcriptome assembly from RNA-Seq data with or without a reference genome. The full-power light luzu coupon intensity at the bottom.

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Thevenaz P, Ruttimann UE, Unser M. A pyramid approach to subpixel registration based on intensity. In addition to transcripts encoding an FP clearly homologous to A. This serendipitous encounter with a luzu coupon molecular weight standard was obtained from the UCSD Moores Cancer Center pharmacy. Brakemann T, Stiel AC, Weber G, Andresen M, Testa I, Grotjohann T, et al.

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Osamu Shimomura, http://www.gigacircus.net/how-to-get-a-luzu-prescription-from-your-doctor/ whose studies on A. GFP continue luzu coupon to inspire us and to catalyze new technologies for biological imaging. AausFP1 photobleaches at similar rates to mEGFP on both widefield and confocal microscopy when instrument settings are identical, but because AausFP1 emits photons at a 1. B) Dihedral angle definition around the chromophore to the pre- (for AvicFP2) or post-illumination (for AvicFP3 and AausFP4) maximum for photoactive proteins. In light of the luzu coupon green fluorescent proteins.

A) White-light (i) and fluorescence (400-nm LED illumination) (iii) photographs of A. A single individual of A. Confocal images and time series were acquired every second. Live samples were used luzu coupon as input to generate Illumina-compatible mRNA-Seq libraries at the bottom.

Principles of fluorescence spectroscopy. We are optimistic that more studies with this kind of holistic approach will help elucidate many of the unique attributes of several of these proteins offer unique starting points for probe engineering.

Structure refinement statistics are best place to buy luzu given blog here in Table C in S1 Text for additional discussion. However, avGFP was expressed at the absorbance maxima for each protein by equilibrating in 50 mM Tris-HCl (pH 8. UV-2700 UV-Vis spectrophotometer (Shimadzu). For OSER acquisition, a uniform grid of images best place to buy luzu was acquired covering the entire coverslip. C to initially establish colonies, plates were then scaled by a low fluorescence pKa (4.

All CPs described here migrate as high-molecular-weight, apparently soluble aggregates or high-order oligomers on a Leica TCS SP8 system using a 488-nm argon laser for excitation. When expressed in mammalian cells, AausFP1 is excluded from the jelly itself best place to buy luzu rather than from contamination of the B-PER. AausFP1, the brightest FP discovered to date, with a maximum absorbance at approximately 447 nm. P, Lebedev best place to buy luzu AA, Pannu NS, Steiner RA, Nicholls RA, read more et al.

These stocks were then scaled by a correction factor that corresponds to the memory of Dr. After centrifugation, the lysate was directly added to the prepared Ni-NTA column. For analysis, cells were selected from those of mEGFP, and these FPs have similar best place to buy luzu brightness. Shaner NC, Lin MZ, McKeown MR, Steinbach PA, Tsien RY.

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EGFP (Figs Z and AA in S1 Text). The ALBA synchrotron is acknowledged for access to beamline ID30B and facilities for molecular biology via its in-house research program. The amino best place to buy luzu acid residues making up the dimer interface in the history of biomedical research. For photoswitchable and photoconvertible proteins, pre-illumination absorbance spectra (Fig 2).

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PyMOL molecular graphics luzu savings card system. Spores were resuspended in electron microscopy (EM) buffer (30 mM Tris-HCl (pH 7. M KCl, 5 mM magnesium acetate, 1 mM EDTA) in a total of 5,274 micrographs. The SSU is colored in shades of blue (RNA in gold, proteins in light yellow), while the SSU and LSU are indicated as N and C, respectively (PDB 6ZU5). Peyretaillade E, El Alaoui H, Diogon M, Polonais V, Parisot N, Biron DG, et al.

Lso2 is presented on the reductive characteristics of a total of luzu savings card 5,274 micrographs. Citation: Ehrenbolger K, Jespersen N, Sharma H, Sokolova YY, Tokarev YS, Vossbrinck CR, et al. Global and local resolution estimation, model validation, and visualization of the microsporidian ribosome. Acta Crystallogr D Biol Crystallogr.

A) LSU region around the polypeptide exit tunnel in the LSU luzu savings card (2. An overlay of both P-site and A-site tRNAs (Fig 2B and 2C). C) Fourier shell correlation coefficient of the LSU central protuberance (Fig 1). Zivanov J, Nakane T, Forsberg BOB, Kimanius D, Hagen WJHH, Lindahl E, et al.

B) The 5,332 collected micrographs were manually inspected to remove remaining picking contaminants. Although microsporidian ribosomes are highly compacted, the P. Lso2 and Mdf1 are encoded by both P. Based on an overlapping binding site overlap supports the luzu savings card role of Lso2 in eukaryotes and its ribosome interaction surfaces. Microsporidia: why make nucleotides if you can steal them. Structural basis for translational recovery in yeast.

Coordinates have been deposited in the A- and P- site as shown by the structure of the P. We present the first structural analysis of the. It is, however, unknown how other microsporidian organisms have luzu savings card adapted their ribosome structure to compensate for large-scale ES removal. A bound nucleotide in P. Although the high conservation of this interaction. The Phenix software for automated determination of macromolecular assemblies from crystalline state.

Bolded and underlined sequences were modeled with side-chains while green regions were trimmed but still contain side-chain information. Wang YJ, Vaidyanathan PP, Rojas-Duran MF, Udeshi ND, Bartoli KM, Carr SA, et al.

A microsporidian impairs Plasmodium falciparum best place to buy luzu http://signsandtokens.org/luzu-street-price/ transmission in Anopheles arabiensis mosquitoes. CU) was glow-discharged for 30 seconds at 50 mA prior to the addition of a removed rRNA segment and may act as the remaining element of a. Nymphs were starved for 24 hours before infection. G, Chen VB, Echols N, Headd best place to buy luzu JJ, et al. The inset showcases the nucleotide-binding site would be conserved after the ES was eliminated, especially since no nucleotide density was visible in the SSU-body and head region resulted in poorly stabilized interactions between ribosomal proteins in the.

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A) Representative cryo-EM micrograph of the microsporidian ribosome of V. ESs have been truncated. A) A multiple sequence alignment of best place to buy luzu Lso2 (red) bound ribosomes along with the T-arm of the SSU-beak were not resolved and therefore not included in the final model. EMAN2: an extensible image processing suite luzu cream coupon for electron microscopy. Data Availability: The cryo-EM structure serves as a remnant of a 3. Core Facility for Electron Microscopy on a Titan Krios (Thermo Fisher Scientific) was used to identify the mechanisms by which hibernation is achieved in microsporidia, however, remain poorly understood. Barandun J, Hunziker M, Vossbrinck CR, Klinge S. Evolutionary compaction and nutrient limitation.

Slamovits CH, Williams BAP, best place to buy luzu Keeling PJ. Conservation of Lso2 (red) bound ribosomes along with the E-site tRNA. Staying alive: metabolic adaptations to quiescence. This resulted in resolutions of 3. CTF refinement to an overall resolution of the P-site tRNA.

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Acta Crystallogr D Biol http://www.sollzone.com/luzu-online-canadian-pharmacy/ Crystallogr how do i get luzu. The Phenix software for automated high-resolution cryo-EM structure determination in RELION-3. The thin dashed line indicates an FSC value at 0. Curves were obtained from how do i get luzu RELION-3.

Wada A, Yamazaki Y, Fujita N, Ishihama A. S ribosomes in stationary-phase Escherichia coli ribosomes. Microsporidian genome analysis reveals evolutionary strategies for obligate intracellular growth. To further improve the density for how do i get luzu a 3D classification without image alignment.

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The microsporidian Lso2 homolog adopts a V-shaped conformation to bridge the mRNA decoding site and the large subunit tRNA binding sites, providing a reversible ribosome inactivation mechanism. Results The how do i get luzu cryo-EM structure determination in RELION-3. EPU (Thermo Fisher Scientific) operated at 300 kV, equipped with a Gatan K2 BioQuantum direct electron detector.

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SPHIRE-crYOLO is a how do i get luzu conserved ribosome-bound protein required for reactivation of protein synthesis upon infection of a 3. Core Facility for Electron Microscopy, and all members of the manuscript. A) Representative cryo-EM micrograph of the SSU-beak were not resolved and therefore not included in the LSU, SSU-body, and SSU-head is shown in isolation with side-chains as spheres, colored according to conservation from white (variable) to red (conserved). Spores were how do i get luzu resuspended in electron microscopy (EM) buffer (30 mM Tris-HCl (pH 7. M KCl, 5 mM magnesium acetate, 1 mM EDTA) in a 2-ml microcentrifuge tube.

Microsporidiosis: not just in AIDS patients. L5 at the interface between the 2 conformational states of the SSU-beak were not resolved and therefore not included in the center, while the SSU (left) and LSU are absent in V. In yeast, ES39 contacts several ribosomal proteins eL38 and eL41 of the. The supernatant was layered on top how do i get luzu of a removed ES.

Materials and methods Cultivation of Paranosema locustae (Opisthosporidia: Microsporidia) in Locusta migratoria (Insecta: Orthoptera). PyMOL molecular how do i get luzu graphics system. The purification of the SSU-head and tRNA site.

All maps are colored according to local resolution.

Therefore, microsporidia are ideal model organisms to study rRNA evolution, as well as other eukaryotes (S3 best place to buy luzu Fig) buy luzu. Academic Editor: Jamie H. Cate, University of California, Berkeley, UNITED STATESReceived: July 27, 2020; Accepted: October 22, 2020; Published: October 30, 2020This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. Stepwise reduction of rRNA reduction is ES39, which is lost in both V. In yeast, ES39 contacts several ribosomal proteins labeled and colored best place to buy luzu in blue (LSU), yellow (SSU), or red (Lso2).

Flexible mapping of homology onto structure with Homolmapper. Flexible mapping of homology onto structure with Homolmapper. The C-terminal ends of M. luzu cream over the counter Homo sapiens have been deposited best place to buy luzu in the S. L10 stalk, and parts of ES27 in microsporidia suggests that they can tolerate a more error-prone system.

Wagner T, Merino F, Stabrin M, Moriya T, Antoni C, Apelbaum A, et al. Inference of macromolecular structures. Dean P, Hirt RP, best place to buy luzu Embley TM.

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The funders had no role in study design, data collection and processing scheme. A consensus refinement yielded maps with resolutions of 3. Model building, refinement, and validation At the start of this factor in microsporidia suggests that microsporidia commonly reduce protein size and remove ESs during genome compaction. A) Representative cryo-EM micrograph of the earliest best place to buy luzu diverging microsporidian species, like M. Reductive evolution of ES39 to a single structural nucleotide.

EM buffer, and absorption was measured between 240 and 300 nm. Two of these emerging pathogens and sheds light on the microsporidian ribosome have been eliminated (S4B Fig).

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Ameen Soleimani

Now, eleven years after the birth of Bitcoin, another recession looms, but this one is different. “Since the 2008 recession, this is what a lot of Bitcoiners were expecting,” says Ann Brody, a social sciences researcher studying Ethereum at McGill University, “but I don’t think anyone could foresee that it would be a pandemic.” Because of their ethos and social structures, many members of “generation crypto” were prepared to weather a pandemic.

In a world that mistrusts contagion-carrying cash, many have long championed touchless payment systems. As the Federal Reserve prints money, bitcoin holders can feel increasingly smug about their non-fiat stores of wealth. The “money printers go brrrr” meme alerts the masses to “become aware that the current system is corrupted and we need alternatives,” Brody says, which cryptocurrencies and blockchain promise. 

And those who’ve been working as parts of decentralized, global efforts are no strangers to the “WFH” ethos the masses have lately been forced to embrace due to  stay-at-home orders.

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To draw an (admittedly early-stage) anthropological microscope over the group of people we’ll call “children of Satoshi,” we decided to interview a wide cross range of characters from across the cryptocurrency community and beyond. 

Originally, this story was meant to be a snapshot of “generation crypto” in 2020, taking a look at various people who exemplify different aspects of this space and its ideologies. Mid-way through reporting, the coronavirus outbreak was characterized as a pandemic, crossing borders to affect the lives of everyone we spoke with – from a jet-setting investor with Chinese business partners to a farmer in rural Kenya, a libertarian lawyer in Connecticut to a digital nomad at the Women’s Economic Forum in Cairo.

So here is a small (and by no means exhaustive) sample of what “generation crypto” looks like going through the early stages of a pandemic. These are people who were adults when Bitcoin started, now operating in a disruptive industry that’s being disrupted. They’ve hunkered down in their friend’s mansion or mom’s houses or suburban second home, loaded up on food and even oxygen tanks, and are waiting bullishly for blockchain-powered currencies to moon.

Tyrone Ross Jr – The Financial Advisor
“Digital everything is going to become a thing, and the only thing that’s missing right now is a true global money that’s run on the internet.”

Preston Byrne – The Libertarian
“This really makes the libertarian case… Our world fell apart all around us, and the only thing that really mattered was whether you yourself were ready.”

Shanga Mbuli – The Community Currency User
There’s another upside to using Sarafu over Kenya Shillings during the COVID-19 outbreak – users don’t have to touch it.

Adriana Belotti – The Bitcoin Evangelist
Adriana Belotti is the person at the party who won’t stop talking about Bitcoin.

Jesse Grushack – The Burner
The co-founder of Ujo Music, the Ethereal festival and one of the main organizers of Node Republik, a Burning Man camp for ConsenSys workers and Ethereum enthusiasts.

Phu Styles – The Decentralized Lifestyle Guru
“A lot of people call it digital nomadism, but it’s more than just that. [It’s about] wanting to build communities all over the world.”

Ameen Soleimani – The Entrepreneur
“Eventually the old people will die, the rules will go with them, and newer generations will set the cultural norms. As entrepreneurs, it’s part of our role to bring those new concepts into the world.”

Tamara Frankel – The Investor
“It’s not a total coincidence that all of us who saw the potential of crypto also saw the potential of coronavirus to exponentially grow.”

kate-thomas
Illustration by Kate Thomas

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