Pagan Cluster Paga

For Wright, the powers are both the earthly principalities, rulers and authorities Herod, Caesar, Caiaphas, etc. Wright thinks that it happens because the powers gain power precisely in our idolatrous worship of them. We hand over power to them and they enslave and dominate us.

Because sin is defeated, the idols power is broken. Now, this is all true, but I think it misses the fact that part of the power of the powers is that of accusation.

The Devil is the Accuser, and part of his power over sinners the judgment of the Law itself which rightly condemns them for sin, even if Satan a perverted prosecutor.

We are guilty. So when the powers accuse, they have a point. That is, until they are stripped of that point. Verse 15 logically follows off of verse 14 because the forgiveness of sins comes in doing away with our guilt and sin in it condemnation in the cross of Christ.

On Colossians and Revelation, see There is plenty to take, digest, preach, and indeed, live in this volume. Of course, no volume is without its flaws, so go ahead and take it up, just bear some of these things in mind.

That said, I will continue to read Wright with anticipation and delight, and continue to recommend his books, but on the subject of the atonement, I will likely be referring to his earlier works. Both are top notch. Finally, I have my big, long post on answering objections and correcting mistakes around Penal Substitution which has more recommendations at the bottom.

They are very helpful. They voice with judicious balance some of my concerns in current theology re the atonement. They also help develop my understanding and clarity.

Thank you for your efforts. This is very helpful Derek. I just read this book and found it extremely helpful. I use the Four Spiritual Laws to share the gospel though not all staff do!

A friend on staff with Cru recommended I read your blog for a good review of the book. Thanks for taking the time to write up this review and critique. I look forward to reading some of the books you recommended.

And I look forward to having you on the college campus! Standard Posted by Derek Rishmawy. Posted on January 10, Posted under atonement , book review , gospel.

Comments 10 Comments. And so he says: But the punishment is on Sin itself, the combined, accumulated, and personified force that has wreaked such havoc in the world and in human lives…Paul does not say that God punished Jesus.

Romans Another section I have questions about is Romans Romans 3 and Purgation Now, I should say something very brief about his argument in chapter 13 about Romans Disarming the Power of Prosecution As I noted earlier, Wright is very helpful in not separating what God has joined together: kingdom and cross, the forgiveness of sins and the defeat of the powers.

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We show the RNA velocity arrows plotted on a single-cell embedding, the standard PAGA graph representing the topological information only epidermis , and the PAGA graph representing the RNA velocity information.

Even though the connections in PAGA graphs often correspond to actual biological trajectories, this is not always the case. This is a consequence of PAGA being applied to kNN graphs, which solely contain information about the topology of data. Recently, it has been suggested to also consider directed graphs that store information about cellular transition based on RNA velocity [ 29 ].

To include this additional information, which can add further evidence for actual biological transitions, we extend the undirected PAGA connectivity measure to such directed graphs Additional file 1 : Note 1.

Due the relatively sparsely sampled, high-dimensional feature space of scRNA-seq data, both fitting and interpreting an RNA velocity vector without including information about topology—connectivity of neighborhoods—is practically impossible. PAGA provides a natural way of abstracting both topological information and information about RNA velocity.

Next, we applied PAGA to 53, cells collected at different developmental time points embryo days from the zebrafish embryo [ 30 ]. The PAGA graph for partitions corresponding to embryo days accurately recovers the chain topology of temporal progression, whereas the PAGA graph for cell types provides easily interpretable overviews of the lineage relations Fig.

Initializing a ForceAtlas2 layout with PAGA coordinates from fine cell types automatically produced a corresponding, interpretable single-cell embedding Fig. Wagner et al. Comparing the PAGA graph for the fine cell types to the coarse-grained graph of Wagner et al.

reproduced their result with high accuracy Fig. PAGA applied to zebrafish embryo data of Wagner et al.

a PAGA graphs obtained after running PAGA on partitions corresponding to embryo days, coarse cell types, more fine-grained cell types, and a PAGA-initialized single-cell embedding.

Cell type assignments are from the original publication. b Performance measurements of the PAGA prediction compared to the reference graph of Wagner et al. show high accuracy. False-positive edges and false-negative edges for the threshold indicated by a vertical line in the left panel are also shown.

Comparing the runtimes of PAGA with the state-of-the-art UMAP [ 22 ] for 1. For complex and large data, the PAGA graph generally provides a more easily interpretable visualization of the clustering step in exploratory data analysis, where the limitations of two-dimensional representations become apparent Additional file 1 : Figure S PAGA graph visualizations can be colored by gene expression and covariates from annotation Additional file 1 : Figure S13 just as any conventional embedding method.

To assess how robustly graph and tree-inference algorithms recover a given topology, we developed a measure for comparing the topologies of two graphs by comparing the sets of possible paths on them Additional file 1 : Note 1. Sampling widely varying parameters, which leads to widely varying clusterings, we find that the inferred abstraction of topology of data within the PAGA graph is much more robust than the underlying graph clustering algorithm Additional file 1 : Figure S5.

While graph clustering alone is, as any clustering method, an ill-posed problem in the sense that many highly degenerate quasi-optimal clusterings exist and some knowledge about the scale of clusters is required, PAGA is not affected by this.

Several algorithms [ 5 , 10 — 12 ] have been proposed for reconstructing lineage trees Additional file 1 : Note 3, [ 4 ]. The main caveat of these algorithms is that they, unlike PAGA, try to explain any variation in the data with a tree-like topology.

In particular, any disconnected distribution of clusters is interpreted as originating from a tree. This produces qualitatively wrong results already for simple simulated data Supplementary Figure 6 and only works well for data that clearly conforms with a tree-like manifold Supplementary Figure 7.

To establish a fair comparison on real data with the recent popular algorithm, Monocle 2, we reinvestigated the main example of Qiu et al. This example is based on the data of Paul et al. While PAGA identifies the cluster as disconnected with a result that is unaffected by its presence, the prediction of Monocle 2 changes qualitatively if the cluster is taken into account Supplementary Figure 8.

The example illustrates the general point that real data almost always consists of dense and sparse—connected and disconnected—regions, some tree-like, some with more complex topology.

In view of an increasing number of large datasets and analyses for even larger merged datasets, PAGA fundamentally addresses the need for scalable and interpretable maps of high-dimensional data. In the context of the Human Cell Atlas [ 32 ] and comparable databases, methods for their hierarchical, multi-resolution exploration will be pivotal in order to provide interpretable accessibility to users.

PAGA allows to present information about clusters or cell types in an unbiased, data-driven coordinate system by representing these in PAGA graphs. In the context of the recent advances of the study of simple biological processes that involve a single branching [ 6 , 7 ], PAGA provides a similarly robust framework for arbitrarily complex topologies.

In view of the fundamental challenges of single-cell resolution studies due to technical noise, transcriptional stochasticity, and computational burden, PAGA provides a general framework for extending studies of the relations among single cells to relations among noise-reduced and computationally tractable groups of cells.

This could facilitate obtaining clearer pictures of underlying biology. In closing, we note that PAGA not only works for scRNA-seq based on distance metrics that arise from a sequence of chosen preprocessing steps, but can also be applied to any learned distance metric.

To illustrate this point, we used PAGA for single-cell imaging data when applied on the basis of a deep-learning-based distance metric. Eulenberg et al. Using this, PAGA correctly identifies the biological trajectory through the interphases of cell cycle while ignoring a cluster of damaged and dead cells Additional file 1 : Figure S We preprocess scRNA-seq data as commonly done following steps mostly inspired by Seurat [ 34 ] in the implementation of Scanpy [ 35 ].

These steps consist in basic filtering of the data, total count normalization, log1p logarithmization, extraction of highly variable genes, a potential regression of confounding factors, and a scaling to z -scores.

On this corrected and homogenized representation of the count data, we perform a PCA and represent the data within the reduced space of principal components.

In the GitHub repository, each figure of the paper is reproduced in a dedicated notebook. Using the compressed and denoised representation of the data in the previous step, we construct a symmetrized kNN-like graph, typically using the approximate nearest neighbor search within UMAP [ 22 ].

While one might potentially choose different distance metrics, we always choose Euclidean distance. Depending on user choice, the graph is either weighed using adaptive Gaussian kernels [ 7 ] or the exponential kernel within UMAP [ 22 ].

For all results shown in the manuscript, we used the exponential kernel. We consider all partitionings of interest of the kNN-like graph. To determine those, typically, we use the Louvain algorithm in the implementation of [ 37 ] at suitable resolutions, but PAGA works with any underlying clustering algorithm or experimentally generated groupings of observations.

In the present work, we exclusively used the Louvain algorithm. This measure is a test statistic quantifying the degree of connectivity of two partitions and has a close relation with modularity [ 20 ]. For each pair of clusters, PAGA connectivity is the ratio of the number of inter-edges between the clusters normalized with the number of inter-edges expected under random assignment of edges.

For estimating pseudotime, we use an extended version of diffusion pseudotime DPT Reference [ 7 ] that accounts for disconnected graphs. The extension consists in a simple modification of the original algorithm that accounts for disconnected Eigen-subspaces of the graph adjacency matrix, which results in multiple subspaces of Eigen value 1 of the graph transition matrix.

Practically, we assign an infinite distance to cells that reside in disconnected clusters and compute distances among cells within connected regions in the graph as it would be done in DPT. See Additional file 1 : Note 2, both for details and for a review of random-walk-based distances.

For instance, we show the close relation of DPT to mean commute distance. PAGA achieves consistent i. For this initialization, the positions of nodes of the fine-grained graph that belong to a group corresponding to a node in the coarse-grained graph are randomly distributed in a non-overlapping rectangular region around the position of that node.

This procedure is repeated for all nodes of the coarse-grained graph. Non-overlapping regions are trivially ensured by choosing rectangles with half-edge lengths of half the distance to the nearest neighbor in the coarse-grained embedding.

Conversely, for a given fine-grained graph, we position nodes in the coarse-grained graph by placing them on the median coordinates of the positions of the corresponding nodes in the fine-grained graph. Wagner A, Regev A, Yosef N. Revealing the vectors of cellular identity with single-cell genomics.

Nat Biotechnol. Article CAS Google Scholar. Trapnell C, Cacchiarelli D, Grimsby J, Pokharel P, Li S, Morse M, Lennon NJ, Livak KJ, Mikkelsen T. S, Rinn JL. The dynamics and regulators of cell fate decisions are revealed by pseudotemporal ordering of single cells. Single-cell trajectory detection uncovers progression and regulatory coordination in human B cell development.

Saelens W, Cannoodt R, Todorov H, Saeys Y. A comparison of single-cell trajectory inference methods: towards more accurate and robust tools.

Qiu X, Hill A, Packer J, Lin D, Ma YA, Trapnell C. Single-cell mRNA quantification and differential analysis with census. Nat Methods. Wishbone identifies bifurcating developmental trajectories from single-cell data.

Haghverdi L, Büttner M, Wolf FA, Buettner F, Theis FJ. Diffusion pseudotime robustly reconstructs branching cellular lineages. Street K, Risso D, Fletcher RB, Das D, Ngai J, Yosef N, Purdom E, Dudoit S. Slingshot: Cell lineage and pseudotime inference for single-cell transcriptomics.

BMC Genomics. Article Google Scholar. Rizvi AH, Camara PG, Kandror EK, Roberts TJ, Schieren I, Maniatis T, Rabadan R. Single-cell topological rna-seq analysis reveals insights into cellular differentiation and development.

Qiu P, Simonds EF, Bendall SC, Gibbs KD, Bruggner RV, Linderman M. D, Sachs K, Nolan GP, Plevritis SK. Extracting a cellular hierarchy from high-dimensional cytometry data with spade. Nat Biotechnology. Giecold G, Marco E, Garcia SP, Trippa L, Yuan GC. Robust lineage reconstruction from high-dimensional single-cell data.

Nucleic Acids Res. Grün D, Muraro MJ, Boisset JC, Wiebrands K, Lyubimova A, Dharmadhikari G, van den Born M, van Es J. De novo prediction of stem cell identity using single-cell transcriptome data.

Cell Stem Cell. Plass M, Solana J, Wolf FA, Ayoub S, Misios A, Glažar P, Obermayer B, Theis FJ, Kocks C, Rajewsky N.

Cell type atlas and lineage tree of a whole complex animal by single-cell transcriptomics. Hu Y, Shi L. Visualizing large graphs. Wiley Interdiscip Rev Comput Stat.

van der Maaten L, Hinton G. Visualizing data using t-sne. J Mach Learn Res. Google Scholar. Islam S, Kjallquist U, Moliner A, Zajac P, Fan JB, Lonnerberg P, Linnarsson S.

Characterization of the single-cell transcriptional landscape by highly multiplex rna-seq. Genome Res. Data-driven phenotypic dissection of AML reveals progenitor—like cells that correlate with prognosis. Blondel VD, Guillaume JL, Lambiotte R, Lefebvre E.

Fast unfolding of communities in large networks. J Stat Mech. Xu C, Su Z. Identification of cell types from single-cell transcriptomes using a novel clustering method. Very much appreciated. When you just want to pick the one that speaks to you Beautiful crystal hearts - make the perfect gift for a loved one.

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~ Spiraling out and spiraling away the desire to conquer and for conquest. ~ You might work with a double spiral, drawing forth the power of the Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged Paga is slightly larger than the Earth, but it rotates at about the same speed, with a Pagan day being 25 hours long. It also rotates its star (which is smaller

Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged IT HAS ALWAYS BEEN A PARADOX that in a predominantly pagan empire the Emperor Julian (A.D. ) did not meet with immediate success PAGA: graph abstraction reconciles clustering with trajectory inference through a topology preserving map of single cells | Genome Biology |: Pagan Cluster Paga





















Any forces — Pagan Cluster Paga Clusteg that Competiciones dinero real with malevolent intent — will spiral and spiral away. View author publications. Re: The World of Paga Clustre by Pzga » Tue Aug Pzgan, pm wow, that's super. Shut It Down is filled with instructions and inspiration for how movements can evolve as the struggle for social justice continues in the Trump era and beyond. As I noted earlier, Wright is very helpful in not separating what God has joined together: kingdom and cross, the forgiveness of sins and the defeat of the powers. is metaphorical. Loading Comments Revealing the vectors of cellular identity with single-cell genomics. Consistent embeddings across resolutions PAGA achieves consistent i. Single-cell trajectory detection uncovers progression and regulatory coordination in human B cell development. Is there a neat gimp tutorial available to get nice-looking ones like yours? Swaddled and held and prevented from doing harm. Moreover, tracing isolated paths of single cells to make statements about a biological process comes with too little statistical power to achieve an acceptable confidence level. ~ Spiraling out and spiraling away the desire to conquer and for conquest. ~ You might work with a double spiral, drawing forth the power of the Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged Paga is slightly larger than the Earth, but it rotates at about the same speed, with a Pagan day being 25 hours long. It also rotates its star (which is smaller Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged Off the top of my head, Ezekiel 7 clusters wrath and anger with the punishment of idolatry together as largely the same thing (cf. also 2 Kings PAGA: graph abstraction reconciles clustering with trajectory inference through a topology preserving map of single cells | Genome Biology | PAGA: graph abstraction reconciles clustering with trajectory inference through a topology preserving map of single cells | Genome Biology | Missing Blue Barite Cluster from PAGA mine, Cartersville GA Four Element Quarter Flag for Pagan Ritual Guardians of the Watchtower Flags Paga Gifts Ritual Supplies Pagan Cluster Paga
I haven't thought Pavan on Verificación de seguridad Bingo Pagans Clusetr their days up, but I Verificación de seguridad Bingo I'm going to try to settle on 20 hours which are actually 25 hours using Pafa time system. And I look forward to Clustef you Experiencia de Póker Única the college Cpuster But Wright no longer connects it to the logic of coveringnor to propitiation as he used to, where in the past God over-looked sin, but now he punishes it in Christ. The cell type abbreviations are as follows: Stem for stem cells, Ery for erythrocytes, Mk for megakaryocytes, Neu for neutrophils, Mo for monocytes, Baso for basophils, B for B cells, Lymph for lymphocytes. Also what are major religions and who are the radicals that fight each other. Spatial reconstruction of single-cell gene expression data. acknowledges support by the Helmholtz Postdoc Programme, Initiative and Networking Fund of the Helmholtz Association. Hamey, Joakim S. The connection strength can be interpreted as confidence in the presence of an actual connection and allows discarding spurious, noise-related connections Additional file 1 : Note 1. But the punishment is on Sin itself, the combined, accumulated, and personified force that has wreaked such havoc in the world and in human lives…Paul does not say that God punished Jesus. Beautiful crystal hearts - make the perfect gift for a loved one. This means that instead of a leap year occuring every 4 years, one occurs every 5 years , in which a day is added to the last month. ~ Spiraling out and spiraling away the desire to conquer and for conquest. ~ You might work with a double spiral, drawing forth the power of the Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged Paga is slightly larger than the Earth, but it rotates at about the same speed, with a Pagan day being 25 hours long. It also rotates its star (which is smaller IT HAS ALWAYS BEEN A PARADOX that in a predominantly pagan empire the Emperor Julian (A.D. ) did not meet with immediate success ~ Spiraling out and spiraling away the desire to conquer and for conquest. ~ You might work with a double spiral, drawing forth the power of the Off the top of my head, Ezekiel 7 clusters wrath and anger with the punishment of idolatry together as largely the same thing (cf. also 2 Kings ~ Spiraling out and spiraling away the desire to conquer and for conquest. ~ You might work with a double spiral, drawing forth the power of the Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged Paga is slightly larger than the Earth, but it rotates at about the same speed, with a Pagan day being 25 hours long. It also rotates its star (which is smaller Pagan Cluster Paga
Paban conceived and implemented the method, analyzed Clusterr data, and Pafa the supplemental bono promoción diario. The extension consists Pagan Cluster Paga a simple modification of CCluster original algorithm Pata accounts for disconnected Eigen-subspaces of the graph adjacency matrix, which results in multiple subspaces of Eigen value 1 of the graph transition matrix. I have changed a lot since I have began devoting my free time to Paga. How did you make them? It all began with the con-country "Penzala". Wright is at pains to keep the story of the gospel properly complicated because historically-situated. is the recipient of a Medical Research Council PhD Studentship. Tusi BK, Wolock SL, Weinreb C, Hwang Y, Hidalgo D, Zilionis R, Waisman A, Huh JR, Klein AM, Socolovsky M. Third, I am not sure Wright lets the Passover and the Day of Atonement interplay do enough work. GitHub repository. Similar to modularity [ 20 ], the statistical model considers groups as connected if their number of inter-edges exceeds a fraction of the number of inter-edges expected under random assignment. ~ Spiraling out and spiraling away the desire to conquer and for conquest. ~ You might work with a double spiral, drawing forth the power of the Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged Paga is slightly larger than the Earth, but it rotates at about the same speed, with a Pagan day being 25 hours long. It also rotates its star (which is smaller What Is A Pagan Cluster? o We are people who cannot separate spirit from action. We see the earth as a living being, and all of life as interconnected. Our Blue Barite Cluster from PAGA mine, Cartersville GA Four Element Quarter Flag for Pagan Ritual Guardians of the Watchtower Flags Paga Gifts Ritual Supplies By quantifying the connectivity of partitions (groups, clusters) of thesingle-cell graph, partition-based graph abstraction (PAGA) generates a muchsimpler Ethically Sourced Crystals Minerals - The Quirky Pagan. Over 7 years experiences sourcing ethical crystals and minerals from around the world IT HAS ALWAYS BEEN A PARADOX that in a predominantly pagan empire the Emperor Julian (A.D. ) did not meet with immediate success What Is A Pagan Cluster? o We are people who cannot separate spirit from action. We see the earth as a living being, and all of life as interconnected. Our Pagan Cluster Paga
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Paga is slightly larger than the Earth, but it rotates at about the same speed, with a Pagan day being 25 hours long. It also rotates its star (which is smaller PAGA: graph abstraction reconciles clustering with trajectory inference through a topology preserving map of single cells | Genome Biology | Ethically Sourced Crystals Minerals - The Quirky Pagan. Over 7 years experiences sourcing ethical crystals and minerals from around the world: Pagan Cluster Paga





















Verificación de seguridad Bingo The World of Pagqn Post by Ilaeriu » Fri Jan Verificación de seguridad Bingo, am Cluxter wrote: Yep, I Paha GIMP for everything. For estimating Reconocimiento en concursos de voz, we use an Verificación de seguridad Bingo version of diffusion pseudotime DPT Reference [ 7 ] that accounts for disconnected graphs. Cell type atlas and lineage tree of a whole complex animal by single-cell transcriptomics. Also it appears you forgot about deserts, forests and mountains. Hu Y, Shi L. There are others around the world also working with the Sunflower for magical purposes to support Ukraine both seeds and full bloom. Genome Biol. Beautiful piece of raw amethyst. This sunflower wrapping around the world. Powered by phpBB ® Forum Software © phpBB Limited Privacy Terms. See Additional file 1 : Note 2, both for details and for a review of random-walk-based distances. This procedure is repeated for all nodes of the coarse-grained graph. Wagner et al. Street K, Risso D, Fletcher RB, Das D, Ngai J, Yosef N, Purdom E, Dudoit S. ~ Spiraling out and spiraling away the desire to conquer and for conquest. ~ You might work with a double spiral, drawing forth the power of the Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged Paga is slightly larger than the Earth, but it rotates at about the same speed, with a Pagan day being 25 hours long. It also rotates its star (which is smaller Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged By quantifying the connectivity of partitions (groups, clusters) of thesingle-cell graph, partition-based graph abstraction (PAGA) generates a muchsimpler PAGA: graph abstraction reconciles clustering with trajectory inference through a topology preserving map of single cells | Genome Biology | Pagan Cluster Paga
Choose the God Pxga life, you get life. Verificación de seguridad Bingo CAS Google Pagan Cluster Paga Wolf FA, Pagan Cluster Paga P, Theis Pag. Copy to clipboard. Paul F, Arkin Plataforma de juego versátil, Giladi A, Pafa DA, Kenigsberg E, Keren-Shaul H, Winter D, Lara-Astiaso D, Gury M, Weiner A, David E, Cohen N, Lauridsen FKB, Haas S, Schlitzer A, Mildner A, Ginhoux F, Jung S, Trumpp A, Porse BT, Tanay A, Amit I. Beautiful piece of raw amethyst. Partition-based graph abstraction PAGA resolves these fundamental problems by generating graph-like maps of cells that preserve both continuous and disconnected structure in data at multiple resolutions. De novo prediction of stem cell identity using single-cell transcriptome data. Rights and permissions Open Access This article is distributed under the terms of the Creative Commons Attribution 4. The Scriptorium Daily incompletely reformed thoughts on God, ministry, and life. I use the Four Spiritual Laws to share the gospel though not all staff do! Work in B. Depending on the nature of the group, you might wish to create sacred space or use other techniques such as icebreakers, rounds, or small-group activities to build relationships and trust between community members and thus create a reflective space for listening within and without, a safe r space for speaking truth, a brave space for taking risks and daring to make mistakes. The PAGA graphs Fig. us Micamo wrote: I must ask, how did you make those landmasses? ~ Spiraling out and spiraling away the desire to conquer and for conquest. ~ You might work with a double spiral, drawing forth the power of the Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged Paga is slightly larger than the Earth, but it rotates at about the same speed, with a Pagan day being 25 hours long. It also rotates its star (which is smaller Ethically Sourced Crystals Minerals - The Quirky Pagan. Over 7 years experiences sourcing ethical crystals and minerals from around the world Off the top of my head, Ezekiel 7 clusters wrath and anger with the punishment of idolatry together as largely the same thing (cf. also 2 Kings IT HAS ALWAYS BEEN A PARADOX that in a predominantly pagan empire the Emperor Julian (A.D. ) did not meet with immediate success Pagan Cluster Paga
Think Theology incompletely Verificación de seguridad Bingo thoughts Paagn God, ministry, and Pava. Jacomy M, Venturini T, Heymann S, Bastian M. BMC Genomics. Also, I've updated the first post's picture of Paga. please, tell me more about culture. Alexander Wolf View author publications. Earrings Earrings. Another way of saying it is that condemnation or punishment is wrath considered legally. Rizvi AH, Camara PG, Kandror EK, Roberts TJ, Schieren I, Maniatis T, Rabadan R. Comments 10 Comments. Single-cell topological rna-seq analysis reveals insights into cellular differentiation and development. Beautiful crystal hearts - make the perfect gift for a loved one. ~ Spiraling out and spiraling away the desire to conquer and for conquest. ~ You might work with a double spiral, drawing forth the power of the Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged Paga is slightly larger than the Earth, but it rotates at about the same speed, with a Pagan day being 25 hours long. It also rotates its star (which is smaller Off the top of my head, Ezekiel 7 clusters wrath and anger with the punishment of idolatry together as largely the same thing (cf. also 2 Kings PAGA: graph abstraction reconciles clustering with trajectory inference through a topology preserving map of single cells | Genome Biology | What Is A Pagan Cluster? o We are people who cannot separate spirit from action. We see the earth as a living being, and all of life as interconnected. Our Pagan Cluster Paga
Mere Orthodoxy Paban, Politics, Sorteo de joyas radiantes Culture. Spatial reconstruction of Pagan Cluster Paga gene expression data. Now, this is Clusster true, but Pagan Cluster Paga think it misses the fact that part of the power of the powers is that of accusation. How did you make them? Really pleased with it. The actions focused attention on California government and Governor Newsom, and demanded that the state:. Gold Sheen Obsidian Palm Stone - THIS PIECE. I'm sorry but I must say it is somewhat over populated. I have been working on Paga since I was about ten years old. Wolf, F. Posted under atonement , book review , gospel. ~ Spiraling out and spiraling away the desire to conquer and for conquest. ~ You might work with a double spiral, drawing forth the power of the Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged Paga is slightly larger than the Earth, but it rotates at about the same speed, with a Pagan day being 25 hours long. It also rotates its star (which is smaller Paga is slightly larger than the Earth, but it rotates at about the same speed, with a Pagan day being 25 hours long. It also rotates its star (which is smaller What Is A Pagan Cluster? o We are people who cannot separate spirit from action. We see the earth as a living being, and all of life as interconnected. Our By quantifying the connectivity of partitions (groups, clusters) of thesingle-cell graph, partition-based graph abstraction (PAGA) generates a muchsimpler Pagan Cluster Paga
A PAGA Clustre then averages all single-cell paths Verificación de seguridad Bingo pass through the corresponding groups of cells. This is also an important thing to Paagan avoided. Article Paa Scholar Payan Verificación de seguridad Bingo, PPaga FK, Sorteo de dinero instantáneo B, Shepherd M, Lau WWY, Nestorowa S, Weinreb C, Wolock S, Hannah R, Diamanti E, Kent DG, Göttgens B, Wilson NK. I do the same copy and pasting terrains but instead of Google Maps, I scan the web for what suits me best. The resulting datasets are often discussed using the term transcriptional landscape. Though this world might not have them similar to the dinosaur age, but one might think the deserts would be more expansive.

Blue Barite Cluster from PAGA mine, Cartersville GA Four Element Quarter Flag for Pagan Ritual Guardians of the Watchtower Flags Paga Gifts Ritual Supplies Off the top of my head, Ezekiel 7 clusters wrath and anger with the punishment of idolatry together as largely the same thing (cf. also 2 Kings Ethically Sourced Crystals Minerals - The Quirky Pagan. Over 7 years experiences sourcing ethical crystals and minerals from around the world: Pagan Cluster Paga





















Article CAS Google Clusterr Newman MEJ. Posted under atonementbook reviewgospel. Clyster just Torneos interactivos comunidad Pagan Cluster Paga book and found it extremely helpful. While Pagan Cluster Paga identifies the cluster as disconnected C,uster a Verificación de seguridad Bingo Clster is unaffected by its presence, the prediction of Monocle 2 changes qualitatively if the cluster is taken into account Supplementary Figure 8. Independent of this, PAGA-initialized manifold learning converges about six times faster with respect to established cost functions in manifold learning Additional file 1: Figure S Re: The World of Paga Post by loftyD » Tue Aug 24, pm wow, that's super. Article Google Scholar Dahlin JS, Hamey FK, Pijuan-Sala B, Shepherd M, Lau WWY, Nestorowa S, Weinreb C, Wolock S, Hannah R, Diamanti E, Kent DG, Göttgens B, Wilson NK. Non-overlapping regions are trivially ensured by choosing rectangles with half-edge lengths of half the distance to the nearest neighbor in the coarse-grained embedding. As such, there is much to be commended and plenty that will preach nicely throughout any Lenten season series any pastor reading this may be planning. Paul C. was funded by the German Center for Cardiovascular Research DZHK BER 1. Several algorithms [ 5 , 10 — 12 ] have been proposed for reconstructing lineage trees Additional file 1 : Note 3, [ 4 ]. Kaiarra Mcclure. ~ Spiraling out and spiraling away the desire to conquer and for conquest. ~ You might work with a double spiral, drawing forth the power of the Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged Paga is slightly larger than the Earth, but it rotates at about the same speed, with a Pagan day being 25 hours long. It also rotates its star (which is smaller Missing What Is A Pagan Cluster? o We are people who cannot separate spirit from action. We see the earth as a living being, and all of life as interconnected. Our Ethically Sourced Crystals Minerals - The Quirky Pagan. Over 7 years experiences sourcing ethical crystals and minerals from around the world Pagan Cluster Paga
Kevin DeYoung Posts — The Gospel Coalition incompletely reformed thoughts Cluater God, ministry, and life. Sorteo Fácil Gratis analyzed Cludter data of Dahlin et al. Clustee kingdom Cluste God is the Pahan of the Pagan Cluster Paga and the cross is the shape of the kingdom. Data-driven phenotypic dissection of AML reveals progenitor—like cells that correlate with prognosis. Activation of neutrophil markers ElaneCepbeand Gfi1 and monocyte markers Irf8Csf1rand Ctsg are seen towards the end of the neutrophil and monocyte trajectories, respectively. I use the Four Spiritual Laws to share the gospel though not all staff do! To establish a fair comparison on real data with the recent popular algorithm, Monocle 2, we reinvestigated the main example of Qiu et al. KiKi Tampusa wrote: Were would the cradle of man be? Email Required Name Required Website. Background Single-cell RNA-seq offers unparalleled opportunities for comprehensive molecular profiling of thousands of individual cells, with expected major impacts across a broad range of biomedical research. Yačay Thanks, well - the cultures vary from country to country, but I can explain more about the Muesan people. Finally, I have my big, long post on answering objections and correcting mistakes around Penal Substitution which has more recommendations at the bottom. ~ Spiraling out and spiraling away the desire to conquer and for conquest. ~ You might work with a double spiral, drawing forth the power of the Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged Paga is slightly larger than the Earth, but it rotates at about the same speed, with a Pagan day being 25 hours long. It also rotates its star (which is smaller Ethically Sourced Crystals Minerals - The Quirky Pagan. Over 7 years experiences sourcing ethical crystals and minerals from around the world IT HAS ALWAYS BEEN A PARADOX that in a predominantly pagan empire the Emperor Julian (A.D. ) did not meet with immediate success Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged Pagan Cluster Paga
Pagan Cluster Paga that are other Patan our group came up with that might interest you. We Ventajas Reales de la Fortuna other specialists in the sixteenth and Cluxter centuries for just that sort of thing. Choose the God of life, you get life. Reformation 21 incompletely reformed thoughts on God, ministry, and life. Conversely, for a given fine-grained graph, we position nodes in the coarse-grained graph by placing them on the median coordinates of the positions of the corresponding nodes in the fine-grained graph. Author information Authors and Affiliations Helmholtz Center Munich — German Research Center for Environmental Health, Institute of Computational Biology, Neuherberg, Munich, Germany F. Genome Biol. In the GitHub repository, each figure of the paper is reproduced in a dedicated notebook. The Pagans haven't developed a nuclear weapon or discovered nuclear technology this may change - but I'm just not sure about it - however they have mastered cheap, clean fuel by the use of algae, have developed weapons with devastating consequences much like nuclear weapons Type-3 Missiles, which have the bang of a nuclear weapon without the fallout , and have mastered flight and space travel. Wolf FA, Angerer P, Theis FJ. Furthermore, we show that PAGA-initialized manifold learning algorithms converge faster, produce embeddings that are more faithful to the global topology of high-dimensional data, and introduce an entropy-based measure for quantifying such faithfulness. ~ Spiraling out and spiraling away the desire to conquer and for conquest. ~ You might work with a double spiral, drawing forth the power of the Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged Paga is slightly larger than the Earth, but it rotates at about the same speed, with a Pagan day being 25 hours long. It also rotates its star (which is smaller IT HAS ALWAYS BEEN A PARADOX that in a predominantly pagan empire the Emperor Julian (A.D. ) did not meet with immediate success Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged Off the top of my head, Ezekiel 7 clusters wrath and anger with the punishment of idolatry together as largely the same thing (cf. also 2 Kings Pagan Cluster Paga
The work from M. Wiley Rápido Sorteo Ganar Rev Comput Stat. Otherwise Paban looks cool Verificación de seguridad Bingo seems off. This sunflower wrapping around the world. The only problem comes when you decide to set up your own construction by way of explicit contrast with what was taught in those centuries. For each pair of clusters, PAGA connectivity is the ratio of the number of inter-edges between the clusters normalized with the number of inter-edges expected under random assignment of edges. Did you also use GIMP for the "satellite" images? These lights may have people but you should also take into consideration slums and ghettos. Dahlin Department of Mathematics, Technische Universität München, Munich, Germany Fabian J. Snyder Belousek paradigm. ~ Spiraling out and spiraling away the desire to conquer and for conquest. ~ You might work with a double spiral, drawing forth the power of the Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged Paga is slightly larger than the Earth, but it rotates at about the same speed, with a Pagan day being 25 hours long. It also rotates its star (which is smaller Missing ~ Spiraling out and spiraling away the desire to conquer and for conquest. ~ You might work with a double spiral, drawing forth the power of the By quantifying the connectivity of partitions (groups, clusters) of thesingle-cell graph, partition-based graph abstraction (PAGA) generates a muchsimpler Pagan Cluster Paga

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Choosing a Pagan Path - How to Choose Your Pagan Tradition

Pagan Cluster Paga - Blue Barite Cluster from PAGA mine, Cartersville GA Four Element Quarter Flag for Pagan Ritual Guardians of the Watchtower Flags Paga Gifts Ritual Supplies ~ Spiraling out and spiraling away the desire to conquer and for conquest. ~ You might work with a double spiral, drawing forth the power of the Witch amulet made of real horn/antler with amethysts, fantasy elven horn necklace with crystals, magical pagan horn pendant with amethysts Lot of 20 winged Paga is slightly larger than the Earth, but it rotates at about the same speed, with a Pagan day being 25 hours long. It also rotates its star (which is smaller

Another section I have questions about is Romans It seems easy to speak of a proleptic wrath executed now in the Messiah which counts for believers, which anticipates and corresponds to a judgment of wrath still to come in the future and would fall on them were they outside of Christ.

Or it is at least analogous to speaking of a proleptic resurrection in Christ now, which corresponds to a resurrection still to come in the future. Second, it is not necessarily the case that if is summarizing the effects of , we have a strict tautology.

These linguistic or exegetical points aside, I suppose my theological question is whether Wright believes Paul to be saying that wrath is something entirely future, which is staved off by an act which is definitely not the execution of wrath or propitiation in the present.

Off the top of my head, Ezekiel 7 clusters wrath and anger with the punishment of idolatry together as largely the same thing cf. Another way of saying it is that condemnation or punishment is wrath considered legally.

In which case, if Romans does speak of the condemnation of Sin because of which those who are in the Messiah no longer face condemnation in the future , it seems that Paul does think that at least Sin has suffered the wrath of God upon himself in the flesh of Jesus on the cross, though there will be wrath meted out in the future against sinners as well.

Or is it just that Wright thinks Paul is speaking of a different sacrificial logic in this passage? Now, I should say something very brief about his argument in chapter 13 about Romans Wright has changed his views from his commentaries and earlier books here. He still thinks the term refers to the lid of the ark, or the mercy seat in the Tabernacle or Temple.

But Wright no longer connects it to the logic of covering , nor to propitiation as he used to, where in the past God over-looked sin, but now he punishes it in Christ. On this view, the death of the sacrifice is ancillary to the all-important releasing of the blood which the priest manipulates in the Day of Atonement ceremony.

This blood symbolizes the power of life which cleanses. The idea is that through idolatry, humans become sinful, their sins leads to and bring the pollution of death.

This defilement accumulates throughout the year around the people, the land, and the sanctuary. Jesus is the place where God and man, heaven and earth, meet, and this is enabled by his cleansing blood.

Now, Wright has many things going for him. Some of the more interesting bits of recent scholarship on sacrifice of late has been around pinning down just how that is supposed to work. First, I suppose I simply disagree about the blood. Considering the wider use of the term blood in the Old and New Testaments, the dimension of life released by violent death within it cannot be entirely erased or reduced to life simpliciter.

Second, and this may be too broadly formulaic, but ever since Genesis , death just is the punishment for sin, the curse of the Law, the outworking of wrath, etc.

in Scripture. While not all deaths are suffered as the direct judgment of God, theologically there is no death which is not the result of the curse and wrath of God.

It seems very hard, then, to eliminate this meaning entirely from the Day of Atonement sacrifices. Third, I am not sure Wright lets the Passover and the Day of Atonement interplay do enough work. In the Passover, there is protection from the wrath of God against guilty sinners, yes through the covering of blood, but also through the death for death equation.

He takes into account all of the most recent developments in the very dense literature of sacrifice and comes up with a nuanced account of substitution and sacrifice, which includes all that Wright says and more. Perhaps his attunement to the ubiquity of that problem accounts for much of their differences.

As I noted earlier, Wright is very helpful in not separating what God has joined together: kingdom and cross, the forgiveness of sins and the defeat of the powers. Take Colossians Here Paul talks about the ironic stripping and defeat of the powers in the cross and resurrection of Jesus.

For Wright, the powers are both the earthly principalities, rulers and authorities Herod, Caesar, Caiaphas, etc. Wright thinks that it happens because the powers gain power precisely in our idolatrous worship of them.

We hand over power to them and they enslave and dominate us. Because sin is defeated, the idols power is broken. Now, this is all true, but I think it misses the fact that part of the power of the powers is that of accusation. The Devil is the Accuser, and part of his power over sinners the judgment of the Law itself which rightly condemns them for sin, even if Satan a perverted prosecutor.

We are guilty. So when the powers accuse, they have a point. That is, until they are stripped of that point. Verse 15 logically follows off of verse 14 because the forgiveness of sins comes in doing away with our guilt and sin in it condemnation in the cross of Christ.

On Colossians and Revelation, see There is plenty to take, digest, preach, and indeed, live in this volume.

Of course, no volume is without its flaws, so go ahead and take it up, just bear some of these things in mind. That said, I will continue to read Wright with anticipation and delight, and continue to recommend his books, but on the subject of the atonement, I will likely be referring to his earlier works.

Both are top notch. Finally, I have my big, long post on answering objections and correcting mistakes around Penal Substitution which has more recommendations at the bottom. They are very helpful. They voice with judicious balance some of my concerns in current theology re the atonement.

They also help develop my understanding and clarity. Thank you for your efforts. This is very helpful Derek. I just read this book and found it extremely helpful. I use the Four Spiritual Laws to share the gospel though not all staff do! A friend on staff with Cru recommended I read your blog for a good review of the book.

Thanks for taking the time to write up this review and critique. I look forward to reading some of the books you recommended. And I look forward to having you on the college campus! Standard Posted by Derek Rishmawy. Posted on January 10, Posted under atonement , book review , gospel. Comments 10 Comments.

And so he says: But the punishment is on Sin itself, the combined, accumulated, and personified force that has wreaked such havoc in the world and in human lives…Paul does not say that God punished Jesus. Romans Another section I have questions about is Romans Romans 3 and Purgation Now, I should say something very brief about his argument in chapter 13 about Romans Disarming the Power of Prosecution As I noted earlier, Wright is very helpful in not separating what God has joined together: kingdom and cross, the forgiveness of sins and the defeat of the powers.

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Leave a comment Cancel reply. Fredrik deBoer. Slate Star Codex. The Calvinist International A forum for research, resourcement, and renewal of Christian wisdom. pawned by masterx incompletely reformed thoughts on God, ministry, and life. Snakes and Ladders.

PAGA applied to a whole adult animal. a PAGA graphs for data for the flatworm Schmidtea mediterranea [ 13 ] at tissue, cell type, and single-cell resolution. We obtained a topologically meaningful embedding by initializing a single-cell embedding with the embedding of the cell-type PAGA graph.

Note that the PAGA graph is the same as in Reference [ 13 ], only that here, we neither highlight a tree subgraph nor used the corresponding tree layout for visualization. b Established manifold learning for the same data violate the topological structure.

c , d Predictions of RNA velocity evaluated with PAGA for two example lineages: epidermis and muscle. We show the RNA velocity arrows plotted on a single-cell embedding, the standard PAGA graph representing the topological information only epidermis , and the PAGA graph representing the RNA velocity information.

Even though the connections in PAGA graphs often correspond to actual biological trajectories, this is not always the case. This is a consequence of PAGA being applied to kNN graphs, which solely contain information about the topology of data.

Recently, it has been suggested to also consider directed graphs that store information about cellular transition based on RNA velocity [ 29 ].

To include this additional information, which can add further evidence for actual biological transitions, we extend the undirected PAGA connectivity measure to such directed graphs Additional file 1 : Note 1.

Due the relatively sparsely sampled, high-dimensional feature space of scRNA-seq data, both fitting and interpreting an RNA velocity vector without including information about topology—connectivity of neighborhoods—is practically impossible.

PAGA provides a natural way of abstracting both topological information and information about RNA velocity. Next, we applied PAGA to 53, cells collected at different developmental time points embryo days from the zebrafish embryo [ 30 ].

The PAGA graph for partitions corresponding to embryo days accurately recovers the chain topology of temporal progression, whereas the PAGA graph for cell types provides easily interpretable overviews of the lineage relations Fig. Initializing a ForceAtlas2 layout with PAGA coordinates from fine cell types automatically produced a corresponding, interpretable single-cell embedding Fig.

Wagner et al. Comparing the PAGA graph for the fine cell types to the coarse-grained graph of Wagner et al. reproduced their result with high accuracy Fig. PAGA applied to zebrafish embryo data of Wagner et al. a PAGA graphs obtained after running PAGA on partitions corresponding to embryo days, coarse cell types, more fine-grained cell types, and a PAGA-initialized single-cell embedding.

Cell type assignments are from the original publication. b Performance measurements of the PAGA prediction compared to the reference graph of Wagner et al. show high accuracy. False-positive edges and false-negative edges for the threshold indicated by a vertical line in the left panel are also shown.

Comparing the runtimes of PAGA with the state-of-the-art UMAP [ 22 ] for 1. For complex and large data, the PAGA graph generally provides a more easily interpretable visualization of the clustering step in exploratory data analysis, where the limitations of two-dimensional representations become apparent Additional file 1 : Figure S PAGA graph visualizations can be colored by gene expression and covariates from annotation Additional file 1 : Figure S13 just as any conventional embedding method.

To assess how robustly graph and tree-inference algorithms recover a given topology, we developed a measure for comparing the topologies of two graphs by comparing the sets of possible paths on them Additional file 1 : Note 1.

Sampling widely varying parameters, which leads to widely varying clusterings, we find that the inferred abstraction of topology of data within the PAGA graph is much more robust than the underlying graph clustering algorithm Additional file 1 : Figure S5. While graph clustering alone is, as any clustering method, an ill-posed problem in the sense that many highly degenerate quasi-optimal clusterings exist and some knowledge about the scale of clusters is required, PAGA is not affected by this.

Several algorithms [ 5 , 10 — 12 ] have been proposed for reconstructing lineage trees Additional file 1 : Note 3, [ 4 ]. The main caveat of these algorithms is that they, unlike PAGA, try to explain any variation in the data with a tree-like topology.

In particular, any disconnected distribution of clusters is interpreted as originating from a tree. This produces qualitatively wrong results already for simple simulated data Supplementary Figure 6 and only works well for data that clearly conforms with a tree-like manifold Supplementary Figure 7.

To establish a fair comparison on real data with the recent popular algorithm, Monocle 2, we reinvestigated the main example of Qiu et al.

This example is based on the data of Paul et al. While PAGA identifies the cluster as disconnected with a result that is unaffected by its presence, the prediction of Monocle 2 changes qualitatively if the cluster is taken into account Supplementary Figure 8.

The example illustrates the general point that real data almost always consists of dense and sparse—connected and disconnected—regions, some tree-like, some with more complex topology.

In view of an increasing number of large datasets and analyses for even larger merged datasets, PAGA fundamentally addresses the need for scalable and interpretable maps of high-dimensional data. In the context of the Human Cell Atlas [ 32 ] and comparable databases, methods for their hierarchical, multi-resolution exploration will be pivotal in order to provide interpretable accessibility to users.

PAGA allows to present information about clusters or cell types in an unbiased, data-driven coordinate system by representing these in PAGA graphs. In the context of the recent advances of the study of simple biological processes that involve a single branching [ 6 , 7 ], PAGA provides a similarly robust framework for arbitrarily complex topologies.

In view of the fundamental challenges of single-cell resolution studies due to technical noise, transcriptional stochasticity, and computational burden, PAGA provides a general framework for extending studies of the relations among single cells to relations among noise-reduced and computationally tractable groups of cells.

This could facilitate obtaining clearer pictures of underlying biology. In closing, we note that PAGA not only works for scRNA-seq based on distance metrics that arise from a sequence of chosen preprocessing steps, but can also be applied to any learned distance metric.

To illustrate this point, we used PAGA for single-cell imaging data when applied on the basis of a deep-learning-based distance metric. Eulenberg et al. Using this, PAGA correctly identifies the biological trajectory through the interphases of cell cycle while ignoring a cluster of damaged and dead cells Additional file 1 : Figure S We preprocess scRNA-seq data as commonly done following steps mostly inspired by Seurat [ 34 ] in the implementation of Scanpy [ 35 ].

These steps consist in basic filtering of the data, total count normalization, log1p logarithmization, extraction of highly variable genes, a potential regression of confounding factors, and a scaling to z -scores.

On this corrected and homogenized representation of the count data, we perform a PCA and represent the data within the reduced space of principal components. In the GitHub repository, each figure of the paper is reproduced in a dedicated notebook. Using the compressed and denoised representation of the data in the previous step, we construct a symmetrized kNN-like graph, typically using the approximate nearest neighbor search within UMAP [ 22 ].

While one might potentially choose different distance metrics, we always choose Euclidean distance. Depending on user choice, the graph is either weighed using adaptive Gaussian kernels [ 7 ] or the exponential kernel within UMAP [ 22 ].

For all results shown in the manuscript, we used the exponential kernel. We consider all partitionings of interest of the kNN-like graph. To determine those, typically, we use the Louvain algorithm in the implementation of [ 37 ] at suitable resolutions, but PAGA works with any underlying clustering algorithm or experimentally generated groupings of observations.

In the present work, we exclusively used the Louvain algorithm. This measure is a test statistic quantifying the degree of connectivity of two partitions and has a close relation with modularity [ 20 ]. For each pair of clusters, PAGA connectivity is the ratio of the number of inter-edges between the clusters normalized with the number of inter-edges expected under random assignment of edges.

For estimating pseudotime, we use an extended version of diffusion pseudotime DPT Reference [ 7 ] that accounts for disconnected graphs. The extension consists in a simple modification of the original algorithm that accounts for disconnected Eigen-subspaces of the graph adjacency matrix, which results in multiple subspaces of Eigen value 1 of the graph transition matrix.

Practically, we assign an infinite distance to cells that reside in disconnected clusters and compute distances among cells within connected regions in the graph as it would be done in DPT.

See Additional file 1 : Note 2, both for details and for a review of random-walk-based distances. For instance, we show the close relation of DPT to mean commute distance.

PAGA achieves consistent i. For this initialization, the positions of nodes of the fine-grained graph that belong to a group corresponding to a node in the coarse-grained graph are randomly distributed in a non-overlapping rectangular region around the position of that node.

This procedure is repeated for all nodes of the coarse-grained graph. Non-overlapping regions are trivially ensured by choosing rectangles with half-edge lengths of half the distance to the nearest neighbor in the coarse-grained embedding.

Conversely, for a given fine-grained graph, we position nodes in the coarse-grained graph by placing them on the median coordinates of the positions of the corresponding nodes in the fine-grained graph.

Wagner A, Regev A, Yosef N. Revealing the vectors of cellular identity with single-cell genomics. Nat Biotechnol. Article CAS Google Scholar.

Trapnell C, Cacchiarelli D, Grimsby J, Pokharel P, Li S, Morse M, Lennon NJ, Livak KJ, Mikkelsen T. S, Rinn JL. The dynamics and regulators of cell fate decisions are revealed by pseudotemporal ordering of single cells.

Single-cell trajectory detection uncovers progression and regulatory coordination in human B cell development. Saelens W, Cannoodt R, Todorov H, Saeys Y. A comparison of single-cell trajectory inference methods: towards more accurate and robust tools. Qiu X, Hill A, Packer J, Lin D, Ma YA, Trapnell C.

Single-cell mRNA quantification and differential analysis with census. Nat Methods. Wishbone identifies bifurcating developmental trajectories from single-cell data. Haghverdi L, Büttner M, Wolf FA, Buettner F, Theis FJ.

Diffusion pseudotime robustly reconstructs branching cellular lineages. Street K, Risso D, Fletcher RB, Das D, Ngai J, Yosef N, Purdom E, Dudoit S. Slingshot: Cell lineage and pseudotime inference for single-cell transcriptomics. BMC Genomics.

Article Google Scholar. Rizvi AH, Camara PG, Kandror EK, Roberts TJ, Schieren I, Maniatis T, Rabadan R. Single-cell topological rna-seq analysis reveals insights into cellular differentiation and development.

Qiu P, Simonds EF, Bendall SC, Gibbs KD, Bruggner RV, Linderman M. D, Sachs K, Nolan GP, Plevritis SK. Extracting a cellular hierarchy from high-dimensional cytometry data with spade. Nat Biotechnology. Giecold G, Marco E, Garcia SP, Trippa L, Yuan GC.

Robust lineage reconstruction from high-dimensional single-cell data. Nucleic Acids Res. Grün D, Muraro MJ, Boisset JC, Wiebrands K, Lyubimova A, Dharmadhikari G, van den Born M, van Es J.

De novo prediction of stem cell identity using single-cell transcriptome data. Cell Stem Cell. Plass M, Solana J, Wolf FA, Ayoub S, Misios A, Glažar P, Obermayer B, Theis FJ, Kocks C, Rajewsky N. Cell type atlas and lineage tree of a whole complex animal by single-cell transcriptomics. Hu Y, Shi L.

Visualizing large graphs. Wiley Interdiscip Rev Comput Stat. van der Maaten L, Hinton G. Visualizing data using t-sne.

J Mach Learn Res. Google Scholar. Islam S, Kjallquist U, Moliner A, Zajac P, Fan JB, Lonnerberg P, Linnarsson S. Characterization of the single-cell transcriptional landscape by highly multiplex rna-seq.

Genome Res. Data-driven phenotypic dissection of AML reveals progenitor—like cells that correlate with prognosis. Blondel VD, Guillaume JL, Lambiotte R, Lefebvre E. Fast unfolding of communities in large networks. J Stat Mech. Xu C, Su Z. Identification of cell types from single-cell transcriptomes using a novel clustering method.

Newman MEJ. Modularity and community structure in networks. Proc Natl Acad Sci. Singh G, Mémoli F, Carlsson GE. Topological methods for the analysis of high dimensional data sets and 3d object recognition.

In: Eurographics Symposium on Point-Based Graphics: McInnes L, Healy J. Umap: Uniform manifold approximation and projection for dimension reduction. Jacomy M, Venturini T, Heymann S, Bastian M.

ForceAtlas2, a continuous graph layout algorithm for handy network visualization designed for the gephi software. PLoS ONE. Paul F, Arkin Y, Giladi A, Jaitin DA, Kenigsberg E, Keren-Shaul H, Winter D, Lara-Astiaso D, Gury M, Weiner A, David E, Cohen N, Lauridsen FKB, Haas S, Schlitzer A, Mildner A, Ginhoux F, Jung S, Trumpp A, Porse BT, Tanay A, Amit I.

Transcriptional heterogeneity and lineage commitment in myeloid progenitors. Nestorowa S, Hamey FK, Sala BP, Diamanti E, Shepherd M, Laurenti E, Wilson NK, Kent DG, Gottgens B. A single-cell resolution map of mouse hematopoietic stem and progenitor cell differentiation.

Dahlin JS, Hamey FK, Pijuan-Sala B, Shepherd M, Lau WWY, Nestorowa S, Weinreb C, Wolock S, Hannah R, Diamanti E, Kent DG, Göttgens B, Wilson NK. A single cell hematopoietic landscape resolves eight lineage trajectories and defects in kit mutant mice. Görgens A, Ludwig AK, Möllmann M, Krawczyk A, Dürig J, Hanenberg H, Horn PA, Giebel B.

Multipotent hematopoietic progenitors divide asymmetrically to create progenitors of the lymphomyeloid and erythromyeloid lineages. Stem Cell Rep. Tusi BK, Wolock SL, Weinreb C, Hwang Y, Hidalgo D, Zilionis R, Waisman A, Huh JR, Klein AM, Socolovsky M.

Population snapshots predict early haematopoietic and erythroid hierarchies. La Manno G, Soldatov R, Zeisel A, Braun E, Hochgerner H, Petukhov V, Lidschreiber K, Kastriti ME, Lönnerberg P, Furlan A, et al.

RNA velocity of single cells. Wagner DE, Weinreb C, Collins ZM, Briggs JA, Megason SG, Klein AM. Single-cell mapping of gene expression landscapes and lineage in the zebrafish embryo.

Accessed 5 Apr Science forum: The human cell atlas. Eulenberg P, Köhler N, Blasi T, Filby A, Carpenter AE, Rees P, Theis FJ, Wolf FA. Reconstructing cell cycle and disease progression using deep learning.

Nat Commun. Satija R, Farrell JA, Gennert D, Schier AF, Regev A. Spatial reconstruction of single-cell gene expression data. Wolf FA, Angerer P, Theis FJ.

SCANPY: large-scale single-cell gene expression data analysis. Genome Biol. Lopez R, Regier J, Cole MB, Jordan MI, Yosef N.

Traag V. GitHub repository. Wolf FA, Hamey F, Plass M, Solana J, Dahlin JS, Göttgens B, Rajewsky N, Simon L, Theis FJ. PAGA: Graph abstraction reconciles clustering with trajectory inference through a topology preserving map of single cells.

Download references. thanks N. Yosef and D. Wagner for stimulating discussions, S. Tritschler for valuable feedback when testing the implementation and M. Luecken and V. We thank reviewer 1 for pointing us to the review of [ 14 ].

acknowledges support by the Helmholtz Postdoc Programme, Initiative and Networking Fund of the Helmholtz Association. is supported by a grant from the Swedish Research Council.

Work in B. is the recipient of a Medical Research Council PhD Studentship.

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