The Xiaomi Mi A1 was hugely popular, with many UK and European buyers importing it from Chinese sellers such as Gearbest and Banggood. Well it looks like Xiaomi have paid attention and will be releasing the successor, the Xiaomi Mi A2 and the A2 Lite in 40 markets with several European countries included.
The Mi A2 will be available in the following European markets: Belarus, Bulgaria, Czech Republic, France, Greece, Hungary, Italy, Poland, Romania, Russia, Slovakia, Spain, Turkey and Ukraine. It's also coming to Colombia and Mexico (no word on North America) and a variety of markets in the Middle East, Africa and Asia Pacific, including Egypt, India, Hong Kong, Philippines, Saudi Arabia, Singapore, Thailand, and the UAE.
Sadly that list does not include the UK, but Xiaomi products have been on Amazon Spain before, so you may still be able to buy it easily.
As we have previously reported the Mi A2 follows the tradition of its predecessor and the Mi A2 is the same hardware as the Xiaomi Mi 6X but running Android One instead of the MIUI 9 version of Android.
The Mi A2 will have an It features a 5.99-inch IPS LCD with a 18:9 aspect ratio and FHD+ resolution of 2160 x 1080 pixels. The screen is tucked under a layer 2.5D Gorilla Glass 3.
The Qualcomm Snapdragon 660 powers the Mi A2 and includes either 4GB of RAM, with 32GB or 64GB of internal storage, or 6GB of RAM with 128GB of space.
The dual camera setup of the Xiaomi Mi A2 includes 20-megapixel Sony IM376 primary sensor with f/1.75 aperture and 2-micron 4-in-1 Super Pixel for capturing brighter shots in dim light conditions. The main sensor is assisted by 12-megapixel Sony IMX486 secondary sensor with 1.25-micron pixel and f/1.75 aperture.
On the front of the phone is a 20-megapixel AI-powered selfie camera for capturing selfie shots with bokeh effect.
There is a 3,000mAh battery which is average at best, and you can charge over USB Type-C.
The cheapest configuration will set you back €249 (roughly £220) in Spain, while the top-tier model, with 6GB of RAM, will cost €349 (roughly £310).
The Xiaomi Mi A2 Lite is actually based on the Redmi 6 Pro and comes with a Qualcomm Snapdragon 625 processor, paired with 3GB of RAM and 32GB of internal storage (€179), or 4GB of RAM and 64GB of space (€229). It will ship with a 5.84-inch, FHD+ display and a larger battery than the Mi A2 4000mAh battery.


The International Centre for Black Sea Studies (ICBSS) hosts the 2nd International Black Sea Symposium on "The Black Sea Region: the State of Play and the Way Forward" to be held on the island of Kalymnos, Dodecanese/Greece, on 30 June - 5 July 2009.
The International Black Sea Symposium (IBSS) provides a forum for study, dialogue and networking in a multicultural and interdisciplinary environment, thus contributing to understanding and cooperation in the Black Sea region and beyond. Young professionals primarily from the countries of the wider Black Sea area, EU member states, the United States and Central Asia are the Symposium target group. They have the opportunity to learn about issues of importance for the future development of the Black Sea region during a four-day intensive course of interactive sessions led by prominent experts. Sessions alternate with conflict resolution workshops guided by expert facilitator Prof. Benjamin Broome of Arizona State University.
Deadline for applications is 31 March 2009.


The course comprises introductory lectures and discussions on key aspects of industrial hygiene and occupational health covering recognition, evaluation and control of health hazards at work. Consideration is given to chemical, physical and biological hazards, and the criteria for the evaluation of each.
Notes: While intended primarily for students planning a career in occupational health, this course provides background to the subject for students studying environmental issues and is strongly recommended for students intending to take ID263.


text:
package org.apache.s2graph.loader.subscriber

import org.apache.hadoop.hbase.client.Put
import org.apache.hadoop.hbase.io.compress.Compression.Algorithm
import org.apache.hadoop.hbase.io.encoding.DataBlockEncoding
import org.apache.hadoop.hbase.mapreduce.{TableOutputFormat}
import org.apache.hadoop.hbase._
import org.apache.hadoop.hbase.regionserver.BloomType
import org.apache.hadoop.hbase.util.Bytes
import org.apache.s2graph.core._
import org.apache.s2graph.core.mysqls.{Label, LabelMeta}
import org.apache.s2graph.core.types.{InnerValLikeWithTs, SourceVertexId, LabelWithDirection}
import org.apache.s2graph.loader.spark.{KeyFamilyQualifier, HBaseContext, FamilyHFileWriteOptions}
import org.apache.s2graph.spark.spark.SparkApp
import org.apache.spark.{SparkContext}
import org.apache.spark.rdd.RDD
import org.hbase.async.{PutRequest}
import play.api.libs.json.Json
import scala.collection.JavaConversions._

object TransferToHFile extends SparkApp with JSONParser {

  val usages =
    s"""
       |create HFiles for hbase table on zkQuorum specified.
       |note that hbase table is created already and pre-splitted properly.
       |
       |params:
       |0. input: hdfs path for tsv file(bulk format).
       |1. output: hdfs path for storing HFiles.
       |2. zkQuorum: running hbase cluster zkQuorum.
       |3. tableName: table name for this bulk upload.
       |4. dbUrl: db url for parsing to graph element.
     """.stripMargin

  //TODO: Process AtomicIncrementRequest too.
  /** build key values */
  case class DegreeKey(vertexIdStr: String, labelName: String, direction: String)

  private def insertBulkForLoaderAsync(edge: Edge, createRelEdges: Boolean = true): List[PutRequest] = {
    val relEdges = if (createRelEdges) edge.relatedEdges else List(edge)
    buildPutRequests(edge.toSnapshotEdge) ++ relEdges.toList.flatMap { e =>
      e.edgesWithIndex.flatMap { indexEdge => buildPutRequests(indexEdge) }
    }
  }

  def buildDegrees(msgs: RDD[String], labelMapping: Map[String, String], edgeAutoCreate: Boolean) = {
    val filtered = msgs.filter { case msg =>
      val tokens = GraphUtil.split(msg)
      tokens(2) == "e" || tokens(2) == "edge"
    }
    for {
      msg <- filtered
      tokens = GraphUtil.split(msg)
      tempDirection = if (tokens.length == 7) "out" else tokens(7)
      direction = if (tempDirection != "out" && tempDirection != "in") "out" else tempDirection
      reverseDirection = if (direction == "out") "in" else "out"
      convertedLabelName = labelMapping.get(tokens(5)).getOrElse(tokens(5))
      (vertexIdStr, vertexIdStrReversed) = (tokens(3), tokens(4))
      degreeKey = DegreeKey(vertexIdStr, convertedLabelName, direction)
      degreeKeyReversed = DegreeKey(vertexIdStrReversed, convertedLabelName, reverseDirection)
      extra = if (edgeAutoCreate) List(degreeKeyReversed -> 1L) else Nil
      output <- List(degreeKey -> 1L) ++ extra
    } yield output
  }
  def buildPutRequests(snapshotEdge: SnapshotEdge): List[PutRequest] = {
    val kvs = GraphSubscriberHelper.g.storage.snapshotEdgeSerializer(snapshotEdge).toKeyValues.toList
    kvs.map { kv => new PutRequest(kv.table, kv.row, kv.cf, kv.qualifier, kv.value, kv.timestamp) }
  }
  def buildPutRequests(indexEdge: IndexEdge): List[PutRequest] = {
    val kvs = GraphSubscriberHelper.g.storage.indexEdgeSerializer(indexEdge).toKeyValues.toList
    kvs.map { kv => new PutRequest(kv.table, kv.row, kv.cf, kv.qualifier, kv.value, kv.timestamp) }
  }
  def buildDegreePutRequests(vertexId: String, labelName: String, direction: String, degreeVal: Long): List[PutRequest] = {
    val label = Label.findByName(labelName).getOrElse(throw new RuntimeException(s"$labelName is not found in DB."))
    val dir = GraphUtil.directions(direction)
    val innerVal = jsValueToInnerVal(Json.toJson(vertexId), label.srcColumnWithDir(dir).columnType, label.schemaVersion).getOrElse {
      throw new RuntimeException(s"$vertexId can not be converted into innerval")
    }
    val vertex = Vertex(SourceVertexId(label.srcColumn.id.get, innerVal))

    val ts = System.currentTimeMillis()
    val propsWithTs = Map(LabelMeta.timeStampSeq -> InnerValLikeWithTs.withLong(ts, ts, label.schemaVersion))
    val labelWithDir = LabelWithDirection(label.id.get, dir)
    val edge = Edge(vertex, vertex, labelWithDir, propsWithTs=propsWithTs)

    edge.edgesWithIndex.flatMap { indexEdge =>
      GraphSubscriberHelper.g.storage.indexEdgeSerializer(indexEdge).toKeyValues.map { kv =>
        new PutRequest(kv.table, kv.row, kv.cf, Array.empty[Byte], Bytes.toBytes(degreeVal), kv.timestamp)
      }
    }
  }

  def toKeyValues(degreeKeyVals: Seq[(DegreeKey, Long)]): Iterator[KeyValue] = {
    val kvs = for {
      (key, value) <- degreeKeyVals
      putRequest <- buildDegreePutRequests(key.vertexIdStr, key.labelName, key.direction, value)
    } yield {
        val p = putRequest
        val kv = new KeyValue(p.key(), p.family(), p.qualifier, p.timestamp, p.value)
        kv
      }
    kvs.toIterator
  }

  def toKeyValues(strs: Seq[String], labelMapping: Map[String, String], autoEdgeCreate: Boolean): Iterator[KeyValue] = {
    val kvs = for {
      s <- strs
      element <- Graph.toGraphElement(s, labelMapping).toSeq if element.isInstanceOf[Edge]
      edge = element.asInstanceOf[Edge]
      putRequest <- insertBulkForLoaderAsync(edge, autoEdgeCreate)
    } yield {
        val p = putRequest
        val kv = new KeyValue(p.key(), p.family(), p.qualifier, p.timestamp, p.value)

        //        println(s"[Edge]: $edge\n[Put]: $p\n[KeyValue]: ${kv.getRow.toList}, ${kv.getQualifier.toList}, ${kv.getValue.toList}, ${kv.getTimestamp}")

        kv
      }
    kvs.toIterator
  }

  override def run() = {
    val input = args(0)
    val tmpPath = args(1)
    val zkQuorum = args(2)
    val tableName = args(3)
    val dbUrl = args(4)
    val maxHFilePerResionServer = args(5).toInt
    val labelMapping = if (args.length >= 7) GraphSubscriberHelper.toLabelMapping(args(6)) else Map.empty[String, String]
    val autoEdgeCreate = if (args.length >= 8) args(7).toBoolean else false
    val buildDegree = if (args.length >= 9) args(8).toBoolean else true
    val compressionAlgorithm = if (args.length >= 10) args(9) else "lz4"
    val conf = sparkConf(s"$input: TransferToHFile")
    conf.set("spark.serializer", "org.apache.spark.serializer.KryoSerializer")
    conf.set("spark.kryoserializer.buffer.mb", "24")

    val sc = new SparkContext(conf)

    GraphSubscriberHelper.management.createTable(zkQuorum, tableName, List("e", "v"), maxHFilePerResionServer, None, compressionAlgorithm)

    /* set up hbase init */
    val hbaseConf = HBaseConfiguration.create()
    hbaseConf.set("hbase.zookeeper.quorum", zkQuorum)
    hbaseConf.set(TableOutputFormat.OUTPUT_TABLE, tableName)
    hbaseConf.set("hadoop.tmp.dir", s"/tmp/$tableName")

    val rdd = sc.textFile(input)

    val kvs = rdd.mapPartitions { iter =>
      val phase = System.getProperty("phase")
      GraphSubscriberHelper.apply(phase, dbUrl, "none", "none")
      toKeyValues(iter.toSeq, labelMapping, autoEdgeCreate)
    }
    //
    //    val newRDD = if (!buildDegree) new HFileRDD(kvs)
    //    else {
    //      val degreeKVs = buildDegrees(rdd, labelMapping, autoEdgeCreate).reduceByKey { (agg, current) =>
    //        agg + current
    //      }.mapPartitions { iter =>
    //        val phase = System.getProperty("phase")
    //        GraphSubscriberHelper.apply(phase, dbUrl, "none", "none")
    //        toKeyValues(iter.toSeq)
    //      }
    //      new HFileRDD(kvs ++ degreeKVs)
    //    }
    //
    //    newRDD.toHFile(hbaseConf, zkQuorum, tableName, maxHFilePerResionServer, tmpPath)
    val merged = if (!buildDegree) kvs
    else {
      kvs ++ buildDegrees(rdd, labelMapping, autoEdgeCreate).reduceByKey { (agg, current) =>
        agg + current
      }.mapPartitions { iter =>
        val phase = System.getProperty("phase")
        GraphSubscriberHelper.apply(phase, dbUrl, "none", "none")
        toKeyValues(iter.toSeq)
      }
    }

    val hbaseSc = new HBaseContext(sc, hbaseConf)
    def flatMap(kv: KeyValue): Iterator[(KeyFamilyQualifier, Array[Byte])] = {
      val k = new KeyFamilyQualifier(CellUtil.cloneRow(kv), CellUtil.cloneFamily(kv), CellUtil.cloneQualifier(kv))
      val v = CellUtil.cloneValue(kv)
      Seq((k -> v)).toIterator
    }
    val familyOptions = new FamilyHFileWriteOptions(Algorithm.LZ4.getName.toUpperCase,
      BloomType.ROW.name().toUpperCase, 32768, DataBlockEncoding.FAST_DIFF.name().toUpperCase)
    val familyOptionsMap = Map("e".getBytes() -> familyOptions, "v".getBytes() -> familyOptions)

    hbaseSc.bulkLoad(merged, TableName.valueOf(tableName), flatMap, tmpPath, familyOptionsMap)
  }

}

redpajama_set_name: RedPajamaGithub


text:
Q: Directional derivative in the boundary of the unit ball in R^3 I have a function $f(r,\theta,\phi)=R(r)\Theta(\theta)\Phi(\phi)$ that is given in spherical coordinates. Consider the set $$\mathbb \Gamma=\left\{(x,y,z)\in \mathbb R^3: x^2+y^2+z^2\leq 1\right\}.$$ I need to calculate $\left.\frac{\partial f}{\partial \boldsymbol\nu}\right|_{\partial \Gamma}$, where $\boldsymbol \nu$ is a unit normal vector to $\partial \Gamma$. I know how to do that in the usual cartesian coordinates but how to compute that derivative without needing to go back to the usual coordinates $(x,y,z)$? I don't know how to represent a unit-vector in $\partial \Gamma$.

A: Your $\Gamma$ is the (closed) unit ball in $\mathbb{R}^3$, so the boundary $\partial\Gamma$ is the surface of said unit ball.
A normal vector to some point $P$ on the surface of a ball lies on the line through $P$ and the origin (or the centre of the ball, in general).
So the normal vector will point in the radial direction, that is in the $\hat{r}$ direction in spherical coordinates.
So want to calculate the directional derivative of $f$ in the $\hat{r}$ direction; in other words, you want to calculate how the value of $f$ changes if we change $r$.
That is just the partial derivative of $f$ with respect to $r$, which in this case is
$$ \frac{\partial f}{\partial r} = R'(r)\Theta(\theta)\Phi(\phi) $$
This is your answer.

A: Note that I am using the mathematics convention for spherical coordinates, i.e $x=r\cos\theta\sin\phi$.
For a unit vector $\mathrm n$ what we mean by $\frac{\partial f}{\partial \mathrm n}$ is actually
$$\nabla f\cdot\mathrm n$$
So,
$$(\nabla f\cdot \mathrm n)|_{\partial\Gamma}=(\nabla f\cdot \mathrm n)(1,\theta,\phi)$$
By normal spherical coordinate rules we know
$$\nabla f=\partial_r f~\mathrm{e}_r+\frac{1}{r\sin\phi}\partial_\theta f~\mathrm e_\theta+\frac{1}{r}\partial_\phi f~\mathrm e_\phi$$
And, simply
$$\mathrm n(r,\theta,\phi)=\mathrm e_r=(1,0,0)$$
Finally, since $\mathrm n$ is a vector and $\nabla f$ is a covector, we can compute the dot product rather simply:
$$\mathrm n\cdot\nabla f=n^i(\nabla f)_i$$
Thus
$$(\nabla f\cdot\mathrm n)(1,\theta,\phi)=(\partial_r f)(1,\theta,\phi)$$
Very simple.

redpajama_set_name: RedPajamaStackExchange


text:
SUGGESTED LEARNING RESOURCES ON RACE AND SYSTEMIC RACISM
The Interfaith Alliance of Oklahoma supports community learning about race and systemic racism. To facilitate this learning, we have compiled a list of books, podcasts, and videos addressing these issues. We hope you will find these resources helpful. We have included many resources to appeal to different needs and interests. Please don't be overwhelmed by the number of items in the list. Pick what interests you. If you don't know where to start, we recommend starting with So You Want to Talk About Race by Ijeoma Oulo.
Coates, Ta-Nehisi (2015). Between the World and Me
Ta-Nehisi Coates is a national correspondent for The Atlantic. His book Between the World and Me won the National Book Award in 2015. Coates is the recipient of a MacArthur Fellowship. He lives in New York City with his wife and son.
"In a profound work that pivots from the biggest questions about American history and ideals to the most intimate concerns of a father for his son, Ta-Nehisi Coates offers a powerful new framework for understanding our nation's history and current crisis. Americans have built an empire on the idea of "race," a falsehood that damages us all but falls most heavily on the bodies of black women and men—bodies exploited through slavery and segregation, and, today, threatened, locked up, and murdered out of all proportion. What is it like to inhabit a black body and find a way to live within it? And how can we all honestly reckon with this fraught history and free ourselves from its burden?" Between the World and Me has won numerous awards."
DiAngelo, Robin J. (2018). White Fragility: Why It's So Hard for White People to Talk About Racism
Robin DiAngelo is an academic, lecturer, and author and has been a consultant and trainer on issues of racial and social justice for more than twenty years. She formerly served as a tenured professor of multicultural education at Westfield State University.
"The New York Times best-selling book exploring the counterproductive reactions white people have when their assumptions about race are challenged, and how these reactions maintain racial inequality."
Kendi, Ibram X. (2019) How to Be an Antiracist
Ibram X. Kendi is a #1 New York Times bestselling author, the Andrew W. Mellon Professor in the Humanities at Boston University, and the founding director of the BU Center for Antiracist Research. Kendi is a contributing writer at The Atlantic and a CBS News correspondent. He will also become the 2020-2021 Frances B. Cashin Fellow at the Radcliffe Institute for the Advanced Study at Harvard University.
"Antiracism is a transformative concept that reorients and reenergizes the conversation about racism—and, even more fundamentally, points us toward liberating new ways of thinking about ourselves and each other. At its core, racism is a powerful system that creates false hierarchies of human value; its warped logic extends beyond race, from the way we regard people of different ethnicities or skin colors to the way we treat people of different sexes, gender identities, and body types. Racism intersects with class and culture and geography and even changes the way we see and value ourselves. In How to Be an Antiracist, Kendi takes readers through a widening circle of antiracist ideas—from the most basic concepts to visionary possibilities—that will help readers see all forms of racism clearly, understand their poisonous consequences, and work to oppose them in our systems and in ourselves. Kendi weaves an electrifying combination of ethics, history, law, and science with his own personal story of awakening to antiracism. This is an essential work for anyone who wants to go beyond the awareness of racism to the next step: contributing to the formation of a just and equitable society."
Oluo, Ijeoma (2019). So You Want to Talk About Race
Ijeoma Oluo is a writer and speaker whose work on race has been featured in the New York Times, Washington Post, Elle, The Guardian, and more. She has twice been named to The Root 100 and received the 2018 Feminist Humanist Award from the American Humanist Society. She lives in Seattle.
"In So You Want to Talk About Race, Ijeoma Oluo guides readers of all races through subjects ranging from intersectionality and affirmative action to "model minorities" in an attempt to make the seemingly impossible possible: honest conversations about race and racism, and how they infect almost every aspect of American life."
Tatum, Beverly Daniel (2017). Why Are All the Black Kids Sitting Together in the Cafeteria?: And Other Conversations about Race (2017)
Beverly Daniel Tatum, Ph.D., is president emerita of Spelman College and in 2014 received the Award for Outstanding Lifetime Contribution to Psychology, the highest honor presented by the American Psychological Association. She lives in Atlanta, Georgia.
"Walk into any racially mixed high school and you will see Black, White, and Latino youth clustered in their own groups. Is this self-segregation a problem to address or a coping strategy? Beverly Daniel Tatum, a renowned authority on the psychology of racism, argues that straight talk about our racial identities is essential if we are serious about enabling communication across racial and ethnic divides. These topics have only become more urgent as the national conversation about race is increasingly acrimonious. This fully revised edition is essential reading for anyone seeking to understand the dynamics of race in America."
Tisby, Jemar and Moore, Lecrae (2019) The Color of Compromise: The Truth About the American Church's Complicity in Racism
Jemar Tisby (B.A., University of Notre Dame, Mdiv Reformed Theological Seminary) is the president of The Witness, a Black Christian Collective where he writes about race, religion, politics, and culture. He is also the co-host of the Pass The Mic podcast. He has spoken nation-wide at conferences and his writing has been featured in the Washington Post, CNN, and Vox. Jemar is a Ph.D. student in History at the University of Mississippi studying race, religion, and social movements in the twentieth century.
Lecrae Devaughn Moore, mononymously known as Lecrae, is an American Christian hip hop recording artist, songwriter, record producer, and actor.
"The Color of Compromise is both enlightening and compelling, telling a history we either ignore or just don't know. Equal parts painful and inspirational, it details how the American church has helped create and maintain racist ideas and practices. You will be guided in thinking through concrete solutions for improved race relations and a racially inclusive church.
The Color of Compromise:
Takes you on a historical, sociological, and religious journey: from America's early colonial days through slavery and the Civil War
Covers the tragedy of Jim Crow laws, the victories of the Civil Rights era, and the strides of today's Black Lives Matter movement
Reveals the cultural and institutional tables we have to flip in order to bring about meaningful integration
Charts a path forward to replace established patterns and systems of complicity with bold, courageous, immediate action
Is a perfect book for pastors and other faith leaders, students, non-students, book clubs, small group studies, history lovers, and all lifelong learners
The Color of Compromise is not a call to shame or a platform to blame white evangelical Christians. It is a call from a place of love and desire to fight for a more racially unified church that no longer compromises what the Bible teaches about human dignity and equality. A call that challenges black and white Christians alike to stand up now and begin implementing the concrete ways Tisby outlines, all for a more equitable and inclusive environment among God's people. Starting today."
Wilkerson, Isabel (2010). The Warmth of Other Suns: The Epic Story of America's Great Migration
Isabel Wilkerson, the winner of the Pulitzer Prize and the National Humanities Medal, is the author of The Warmth of Other Suns, the New York Times bestseller that tells the story of the Great Migration, a watershed in American history. Her second book, Caste: The Origins of Our Discontents, is forthcoming from Random House in August 2020. For The Warmth of Other Suns, Wilkerson won the National Book Critics Circle Award for Nonfiction, the Heartland Prize for Nonfiction, the Anisfield-Wolf Award for Nonfiction, the Lynton History Prize from Harvard and Columbia universities, the Stephen Ambrose Oral History Prize and was shortlisted for both the Pen-Galbraith Literary Award and the Dayton Literary Peace Prize.
"From 1915 to 1970, this exodus of almost six million people changed the face of America. Wilkerson compares this epic migration to the migrations of other peoples in history. She interviewed more than a thousand people, and gained access to new data and official records, to write this definitive and vividly dramatic account of how these American journeys unfolded, altering our cities, our country, and ourselves.
With stunning historical detail, Wilkerson tells this story through the lives of three unique individuals: Ida Mae Gladney, who in 1937 left sharecropping and prejudice in Mississippi for Chicago, where she achieved quiet blue-collar success and, in old age, voted for Barack Obama when he ran for an Illinois Senate seat; sharp and quick-tempered George Starling, who in 1945 fled Florida for Harlem, where he endangered his job fighting for civil rights, saw his family fall, and finally found peace in God; and Robert Foster, who left Louisiana in 1953 to pursue a medical career, the personal physician to Ray Charles as part of a glitteringly successful medical career, which allowed him to purchase a grand home where he often threw exuberant parties.
Wilkerson brilliantly captures their first treacherous and exhausting cross-country trips by car and train and their new lives in colonies that grew into ghettos, as well as how they changed these cities with southern food, faith, and culture and improved them with discipline, drive, and hard work. Both a riveting microcosm and a major assessment, The Warmth of Other Suns is a bold, remarkable, and riveting work, a superb account of an "unrecognized immigration" within our own land. Through the breadth of its narrative, the beauty of the writing, the depth of its research, and the fullness of the people and lives portrayed herein, this book is destined to become a classic."
What's CODE SWITCH? It's the fearless conversations about race that you've been waiting for! Hosted by journalists of color, this NPR podcast tackles the subject of race head-on.
5 Podcasts To Listen To If You Really Want To Know About Race in America - From WBUR
My podcast episode From Katie Couric with Jamie Foxx, Michael B. Jordan, and Bryan Stevenson about Just Mercy
Still Processing, a New York Times culture podcast with Jenna Wortham and Wesley Morrison
Seeing White - a Scene on the Radio podcast
Jemele Hill is Unbothered, a podcast with award-winning journalist Jemele Hill
Hear To Slay, "the black feminist podcast of your dreams," with Roxane Gay and Tressie McMillan Cottom
Pod Save The People, organizer and activist DeRay McKesson explores news, culture, social justice, and politics with analysis from fellow activists Brittany Packnett, Sam Sinyangwe, and writer Dr. Clint Smith III
The Appeal, a podcast on criminal justice reform hosted by Adam Johnson
Justice In America, a podcast by Josie Duffy Rice and Clint Smith on criminal justice reform
Brené Brown with Ibram X. Kendi, a podcast episode on antiracism
Come Through, a WNYC podcast with Rebecca Carroll
The Kinswomen, conversations on race, racism, and allyship between women, hosted by Hannah Pechter and Yseult Polfliet
DOCUMENTARIES, VIDEOS AND MOVIES
White Fragility: Why It's So Hard for White People to Talk About Racism by Robin DiAngelo
Dr. Robin DiAngelo is an academic, lecturer, and author and has been a consultant and trainer on issues of racial and social justice for more than twenty years. Drawing on more than twenty years of research, Dr. DiAngelo helps white people get out of denial and develop their capacity to engage more constructively about race.
13th By Ava DuVernay. This is the full-length documentary that Netflix made available to everyone. Combining archival footage with testimony from activists and scholars, director Ava DuVernay's examination of the U.S. prison system looks at how the country's history of racial inequality drives the high rate of incarceration in America.
Let's get to the root of racial injustice | Megan Ming Francis| TEDxRainier. Megan Ming Francis is an Assistant Professor in the Department of Political Science at the University of Washington where she specializes in the study of American politics, race, and the development of constitutional law. She is particularly interested in the construction of rights and citizenship, black political activism, and the post-civil war South. Born and raised in Seattle, WA, she was educated at Garfield High School, Rice University in Houston, and Princeton University where she received her M.A. and her Ph.D. in Politics. In this inspiring and powerful talk, Megan Francis traces the root causes of our current racial climate to their core causes, debunking common misconceptions and calling out "fix-all" cures to a complex social problem
Black in America: And Still I Rise Henry Louis Gates, Jr. In his four-hour series, Henry Louis Gates, Jr. embarks on a deeply personal journey through the last fifty years of African American history. Joined by leading scholars, celebrities, and a dynamic cast of people who shaped these years, Gates travels from the victories of the civil rights movement up to today, asking profound questions about the state
Racism and the Soul of America – Eddie Gaude 2016 lecture. Dr. Eddie Glaude, Jr, is a Professor of Religion and African-American Studies at Princeton University. Dr. Glaude speaks very frankly about systemic racism and challenges the viewers to both understand and move into action.
Kendi, Ibram X. How to be an Antiracist
Dr. Kendi at the Aspen Institute
Dr. Kendi on the Colbert Show. Approximately 15 minutes long. Antiracism as different from not racist…
A TED talk with Dr. Kendi in conversation with two others to clarify the difference between "not racist" and "antiracist."
Ibram X. Kendi is an American author, historian, and scholar of race and discriminatory policy in America. In July 2020, he assumed the position of director of the Center for Antiracist Research at Boston University. His work in Boston is a continuation of his work at the Antiracist Research and Policy Center at the American University.

redpajama_set_name: RedPajamaCommonCrawl


text:
The utility of pathway selective estrogen receptor ligands that inhibit nuclear factor-κB transcriptional activity in models of rheumatoid arthritis
James C Keith Jr1,
Leo M Albert1,
Yelena Leathurby1,
Max Follettie2,
Lili Wang2,
Lisa Borges-Marcucci3,
Christopher C Chadwick4,
Robert J Steffan5 &
Douglas C Harnish3
Arthritis Research & Therapy volume 7, Article number: R427 (2005) Cite this article
Rheumatoid arthritis (RA) is a chronic inflammatory disease that produces synovial proliferation and joint erosions. The pathologic lesions of RA are driven through the production of inflammatory mediators in the synovium mediated, in part, by the transcription factor NF-κB. We have identified a non-steroidal estrogen receptor ligand, WAY-169916, that selectively inhibits NF-κB transcriptional activity but is devoid of conventional estrogenic activity. The activity of WAY-169916 was monitored in two models of arthritis, the HLA-B27 transgenic rat and the Lewis rat adjuvant-induced model, after daily oral administration. In both models, a near complete reversal in hindpaw scores was observed as well as marked improvements in the histological scores. In the Lewis rat adjuvant model, WAY-169916 markedly suppresses the adjuvant induction of three serum acute phase proteins: haptoglobin, α1-acid glycoprotein (α1-AGP), and C-reactive protein (CRP). Gene expression experiments also demonstrate a global suppression of adjuvant-induced gene expression in the spleen, liver, and popliteal lymph nodes. Finally, WAY-169916 was effective in suppressing tumor necrosis factor-α-mediated inflammatory gene expression in fibroblast-like synoviocytes isolated from patients with RA. Together, these data suggest the utility of WAY-169916, and other compounds in its class, in treating RA through global suppression of inflammation via selective blockade of NF-κB transcriptional activity.
Rheumatoid arthritis (RA) is a chronic, debilitating condition affecting 0.5 to 1% of the world's population. The major goals of treatment of RA are to reduce pain and discomfort, to prevent deformities and loss of joint function, and to maintain a productive and active lifestyle. RA is characterized by chronic joint inflammation mediated by inflammatory cell infiltration into synovial tissues as well as joint destruction through the overexpression of matrix metalloproteinase (MMP) in articular synoviocytes and chondrocytes. The pathologic lesions of RA are driven, in part, by the production of inflammatory mediators in synoviocytes and macrophages, probably involving the transcription factor NF-κB. Because NF-κB is localized in the nuclei of synovial cells in patients with RA [1, 2] and the inducers and targets of NF-κB almost perfectly match the list of pivotal mediators increased in RA [3], an important role for activated NF-κB in human RA is likely.
NF-κB is a dimeric transcription factor composed of homodimeric and heterodimeric complexes of the Rel family of proteins, p65 (Rel A), p50/105, c-Rel, p52/100, and Rel B. Binding of cytoplasmic inhibitory protein-κB (IκB) to NF-κB masks the NF-κB nuclear localization signal and sequesters NF-κB in a non-activated form in the cytoplasm. Cell activation by a variety of extracellular signals such as oxidative stress, cytokines, and lipopolysaccharide induces a cascade of events that leads to the degradation of IκB; activated NF-κB then translocates to the nucleus, where it binds to DNA elements in the promoters of several proinflammatory gene families [4].
Activation of NF-κB has been observed in synovial cells from patients with RA [5] and results in the induction of proinflammatory genes such as tumor necrosis factor-α (TNF-α), IL-1β, IL-6, MMP-1, and MMP-3 in ex vivo synovial membrane cultures [6]. Moreover, NF-κB activation might also be a pivotal factor protecting cells from apoptosis, thus contributing to synovial hyperplasia (reviewed in [7]). Inactivation of NF-κB in transgenic mice expressing a 'super-repressor' IκBα or in rel- /- and nfkb1- /- knockout mice rendered the animals refractory to development of collagen-induced arthritis [8, 9]. In another study performed in the rat adjuvant-induced arthritis model, intra-articular injection of an adenoviral construct encoding a dominant-negative from of IκB kinase-2 significantly ameliorated the severity of the adjuvant arthritis and was correlated with a decrease in NF-κB DNA binding in the nucleus of synovial cells [10]. Because NF-κB is involved in normal immune and homeostatic processes, its prolonged inhibition might be harmful. Therefore, more indirect methods of targeting NF-κB might provide a safer pharmacological profile.
In tissues that express estrogen receptor (ER), 17β-estradiol inhibits NF-κB-driven transcription through multiple mechanisms that might include direct protein–protein interactions [11, 12], inhibition of NF-κB binding to DNA [13, 14], induction of IκB expression [15], or coactivator sharing [16, 17]. Two nuclear estrogen receptors have been identified (ERα and ERβ). Both receptors are widely distributed throughout numerous organs [18] and are present in T cells, monocytes, dendritic cells, synovial macrophages, articular chondrocytes, and proliferating fibroblasts present in the RA joint [19–22]. These two receptors have a nearly identical DNA-binding domain, both activate transcription through binding to identical ER response elements [23, 24], and both can antagonize NF-κB transcriptional activity [25, 26]. Taken together, these findings identify RA as a disease amenable to treatment with ER-selective NF-κB inhibitors.
The selective inflammatory modulator WAY-169916 is a non-steroidal ER-dependent inhibitor of NF-κB transcriptional activity. Although it inhibits the expression of a range of inflammatory proteins, including cytokines, chemokines, and cell adhesion molecules that are expressed after activation of NF-κB, WAY-169916 lacks estrogenic activity such as the stimulation of uterine proliferation [27]. Here we demonstrate that WAY-169916 is efficacious in two models of arthritis, the HLA-B27 transgenic rat and a Lewis rat model of adjuvant-induced arthritis. The activity of WAY-169916 is related to its ability to suppress inflammatory processes globally, as demonstrated by the decrease in serum acute-phase protein levels of haptoglobin, α1-acid glycoprotein (α1-AGP), and C-reactive protein (CRP) as well as the inhibition of adjuvant-induced gene expression in the spleen, liver, and popliteal lymph nodes in the rat adjuvant arthritis model. Moreover, WAY-169916 was also active in suppressing cytokine and adhesion molecule expression in fibroblast-like synoviocytes (FLS) isolated from patients with RA. Taken together, these data suggest the potential utility of the pathway-selective ER ligand WAY-169916 and other compounds in its class in the treatment of RA.
Male HLA-B27 transgenic rats were obtained from Taconic; the Lewis rats were purchased from Charles River Laboratories. The rats were housed in accordance with standard operating procedures and were provided with food and water ad libitum. All experiments were approved and performed in accordance with the Wyeth Animal Care and Use Committee standards.
HLA-B27 transgenic rat model arthritis
HLA-B27 transgenic rats, 26 to 28 weeks old, experiencing maximal clinical signs of arthritis with a score of 12, using a scale of 0 to 3 for swelling and for erythema of the hindpaws (0, normal paw; 1, mild; 2, moderate; 3, severe) were treated with vehicle (2% Tween 80, 0.5% methylcellulose), prednisolone (0.6 mg/kg), or WAY-169916 (10 mg/kg) given orally once daily for 29 days with four rats per group. At necropsy, the tarsal joints were removed and prepared for histological examination. After decalcification, histological sections were stained with hematoxylin and eosin or Safranin O/Fast Green stain. Synovial tissue from tarsal joints was evaluated on the basis of synovial hyperplasia, fibroplasia, inflammatory cell infiltration, and pannus formation [28].
Articular cartilage was evaluated with Mankin's histological grading system [29]. The scoring system evaluates the structure of the articular cartilage, ranging from 0 (normal), 1 (surface irregularity), 2 (pannus and surface irregularity), 3 (clefts to transititional zone), 4 (clefts to radial zone), 5 (clefts to the calcified zone), to 6 (complete disorganization; cartilage cells, ranging from 0 (normal), 1 (diffuse hypercellularity), 2 (cloning), to 3 (hypocellularity); Safranin-O staining to assess proteoglycan content, ranging from 0 (normal), 1 (slight reduction), 2 (modest reduction), 3 (severe reduction), to 4 (no staining); and tidemark integrity, ranging from 0 (intact) to 1 (crossed by blood vessels). The scores for eachtarsal joint were tabulated and summed, and amean score was derived for each animal, ranging from 0 to 14. Statistical analysis was performed with Abacus Concepts Super ANOVA (Abacus Concepts, Inc., Berkeley, CA). All parameters of interest were subjected to analysis of variance (ANOVA) with Duncan's new multiple-range post hoc testing between groups. Data are expressed throughout as means ± standard deviation (SD), and differences were deemed significant if P < 0.05.
Rat adjuvant-induced arthritis model
Arthritis was induced in the Lewis rats with complete Freund's adjuvant (CFA) by intradermal injection of 0.1 mg of heat-killed and dried Mycobacterium tuberculosis in 0.1 ml of mineral oil, at the base of the tail. Eight days after adjuvant injection, when the rats were experiencing maximal clinical signs of arthritis with a score of 12 using the same hindpaw scoring system described above, treatment began. Male Lewis rats (n = 6) received orally delivered vehicle (2.0% Tween 80, 0.5% methylcellulose, 1 ml/kg) or WAY-169916 (5.0, 0.3, or 0.1 mg/kg) once daily for 10 to 14 days, with six rats in each group. The clinical signs of arthritis were monitored daily. At the end of the experiment, terminal blood samples were obtained and the tarsal joints were prepared for histological examination and graded as described above. Statistical analysis was performed with Abacus Concepts Super ANOVA. All parameters of interest were subjected to ANOVA with Duncan's new multiple-range post hoc testing between groups. The serum samples were used to determine the levels of haptoglobin, α1-AGP, and CRP by radial immunodiffusion test kits in accordance with manufacturer's protocol (Life Diagnostics Inc.). The data were analyzed by one-way ANOVA and are expressed as means ± SD, and differences were deemed significant if P < 0.05.
Gene expression profiling experiments were conducted with RNA isolated from the spleen, liver, and popliteal lymph nodes, using Affymetrix REA230A oligonucleotide arrays (Affymetrix) in accordance with the manufacturer's recommendations. The arrays were washed and stained with Streptavidin R–phycoerythrin (Molecular Probes) with the use of the GeneChip® Fluidics Station 400, and scanned with a Hewlett Packard GeneArray Scanner in accordance with the manufacturer's instructions. Fluorescent data were collected and converted to gene-specific difference averages with MicroArray Suite 4.0 software. A representative set of genes regulated by WAY-169916 was confirmed by real-time RT–PCR analysis. All mRNA levels were normalized for glyceraldehyde-3-phosphate dehydrogenase expression. The data were analyzed by one-way ANOVA and expressed as means ± SD, and differences were deemed significant if P < 0.05.
NF-κB DNA binding experiments
Mouse splenocytes were prepared by creating single-cell suspensions, with the subsequent removal of red blood cells with Tris-NH4Cl solution. After lysis of red blood cells, the cells were cultured in 24-well plates at a concentration of 106 cells/ml in RPMI-10 (RPMI medium containing 10% heat-inactivated fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin, 2 mM glutamine, and 50 μM 2-mercaptoethanol; Invitrogen, Carlsbad, CA). The cells were stimulated with concanavalin A (ConA) and co-treated with either WAY-169916 (1 μM) or pyrrolidine dithiocarbamate (PDTC; 100 μM) for 18 hours. Nuclear extract preparation and NF-κB DNA binding experiments were conducted with kits purchased from Active Motif.
Experiments with FLS
Human FLS isolated from patients with RA were purchased from Cell Applications, Inc. The cells were cultured in synoviocyte growth medium (Cell Applications, Inc.) and seeded at 3 × 105 cells per well in a 12-well dish. After overnight culture, the cells were pretreated for 1 hour with vehicle, WAY-169916 (1 μM), or PDTC (100 μM), followed by stimulation for 1 hour with TNF-α (100 U/ml). Synoviocyte RNA was isolated after the 1 hour of TNF-α treatment, and gene expression analysis was performed using real-time RT–PCR with an ABI PRISM 7900 Sequence Detection System, in accordance with the manufacturer's protocol (Applied Biosystems). The data were analyzed with Sequence Detector v2.1 software (Applied Biosytems) and normalized to glyceraldehyde-3-phosphate dehydrogenase with the Applied Biosystems primer set. Values are reported as means ± SEM for each group from two experiments, with n = 3. The data were analyzed by one-way ANOVA and differences were deemed significant if P < 0.05.
Activity in the hla-b27 transgenic rat
The HLA-B27 transgenic rat expresses two human proteins (HLA-B27 and β2-microglobulin) that, over time, provoke a misdirected immune response. This model represents a chronic intestinal inflammation with associated arthritis induced by the human class I major histocompatibility allele HLA-B27, which is strongly associated with human disease. Treatment of male HLA-B27 transgenic rats with concentrations of WAY-169916 as low as 0.05 mg/kg rapidly converts the chronic diarrhea that these rats experience to a normal stool [27]. If the disease is allowed to progress, they begin to show symptoms of arthritis. In these settings, treatment of WAY-169916 at a single oral dosage of 10 mg/kg per day restored the clinical joint scores to baseline after 10 days, while a sub-optimal dose of prednisolone (0.6 mg/kg) resulted in a 50% improvement in the joint scores (Fig. 1). Histological scoring of synovitis and cartilage damage in the tarsal joints after 29 days of treatment was also conducted. Treatment with WAY-169916 significantly decreased the synovitis parameters of synovial structure, fibroplasia, inflammatory cell infiltrates, and total synovitis score, and also significantly improved all cartilage parameters monitored (Table 1).
WAY-169916 improves joint scores in HLA-B27 transgenic rat model of arthritis. HLA-B27 transgenic rats, 26 to 28 weeks old, presenting signs of arthritis were treated orally daily with vehicle, prednisolone (0.6 mg/kg), or WAY-169916 (10 mg/kg) for 29 days. Joint scores were assessed by evaluating hindpaws for erythema and swelling (0 to 3 each; maximal score of 12).
Table 1 Histological scoring of synovitis and cartilage damage in the tarsal joints from HLA-B27 transgenic rats
Activity of WAY-169916 in the Lewis rat adjuvant-induced arthritis model
WAY-169916 was then given a more thorough evaluation in the male Lewis rat adjuvant-induced arthritis model. The disease in this model is a migratory polyarthritis affecting primarily the tarsal, metatarsal, and interphalangeal joints. The hallmarks of the model include polyarticular inflammation, marked bone resorption, and periosteal bone proliferation. When immunized with CFA, the joints of Lewis rats swell markedly over a period of 8 days. After maximal swelling had occurred, rats received an oral daily dose of WAY-169916, making this a therapeutic dosing regimen. Joint swelling was rapidly and markedly reduced in rats treated with WAY-169916. Full efficacy was seen with oral doses of 0.3 mg/kg or higher (Fig. 2) but efficacy was decreased at a dose of 0.1 mg/kg. However, both doses were effective at reversing tarsal joint destruction as assessed by synovitis and cartilage (Mankin) scores (Table 2). Incremental improvements in the histology scores were observed with higher doses of WAY-169916 (data not shown), suggesting that continued improvements in joint lesions might occur with a longer duration of treatment or with higher dosages.
WAY-169916 improves joint scores in a dose-dependent fashion in rat adjuvant-induced arthritis model. (a) Male Lewis rats were injected with complete Freund's adjuvant on day 1 and maximal inflammation was allowed to develop. Beginning on day 8 and continuing until day 22, rats were treated daily with oral vehicle or WAY-169916 (0.3 and 0.1 mg/kg). Joint scores were assessed by evaluating hindpaws for erythema and swelling (0 to 3 each; maximal score of 12). (b) WAY-169916 improves joint scores in rat adjuvant-induced arthritis model in both males and females. Experiments were performed as in (a) except that WAY-169916 was dosed daily at 5 mg/kg orally in both intact male and intact female rats.
Table 2 Histological scoring of synovitis and cartilage changes in the tarsal joints from rats with adjuvant-induced arthritis
Because both the HLA-B27 transgenic rat and Lewis rat studies used males, the efficacy of WAY-169916 (5 mg/kg) was compared in intact male and female Lewis rats with the same experimental design as described above. The joint (Fig. 2b) and histology scores (not shown) for the two sexes were equivalent; it therefore does not seem that the utility of WAY-169916 is restricted by gender.
Mechanism of action of WAY-169916
Because WAY-169916 has been shown to antagonize NF-κB transcriptional activity selectively [27], we wished to begin to address how WAY-169916 might be functioning to improve disease symptoms in the rat adjuvant model. Previous studies have shown changes in concentrations of rat serum proteins induced by adjuvant administration (reviewed in [30]). We decided to look at three acute-phase proteins, haptoglobin, α1-AGP, and CRP, that are induced by the adjuvant and have been correlated with RA progression in humans [31]. Serum was analyzed from male Lewis rats treated with 5 mg/kg WAY-169916 for 10 days. As shown in Fig. 3, both haptoglobin and α1-AGP serum levels were induced about 300 to 400% by adjuvant treatment, whereas CRP inductions were more modest (40%); this was consistent with previous reports [30]. WAY-169916 inhibited the adjuvant induction of all three acute-phase proteins but had no effect on their basal levels.
WAY-169916 inhibits the adjuvant-induced expression of serum acute-phase protein. Serum from control male rats or adjuvant-induced rats treated with either vehicle or WAY-169916 (5 mg/kg) was analyzed for the expression of (a) haptoglobin, (b) α1-acid glycoprotein (α1-AGP) or (c) C-reactive protein (CRP) by radial immunodiffusion assay. Results are expressed as means ± SEM from six rats per group. *P < 0.05 compared with vehicle control. CFA, complete Freund's adjuvant.
We also performed gene expression profile analysis from the spleen, liver, and popliteal lymph nodes from these rats. In the spleen, 36 genes were identified that were induced twofold by adjuvant treatment (average fold change; Table 3). Of those 36 genes, WAY-169916 decreased the expression of 29 of them by at least 50%. Several genes that have been implicated in the pathogenesis of RA that were regulated by WAY-169916 include LBS binding protein (LBP), CD14, MMP-9, IL1R2, S100A8, and S100A9. As a control, the regulation of LBP, haptoglobin, and S100A9 was confirmed by real-time RT–PCR (Fig. 4a). A similar global inhibition of adjuvant-induced genes by WAY-169916 was also observed in liver and popliteal lymph node gene-profiling studies. In the liver, 47 genes were induced and WAY-169916 inhibited 43 of those by 50%; in the lymph node, 143 genes were induced and 61 of those were repressed by 50% by WAY-169916 (data not shown).
Table 3 WAY-169916 gene-profiling experiment with spleen from rat adjuvant arthritis model
The effect of WAY-169916 in spleen cells. (a) The regulation of LBS binding protein (LBP), haptoglobin and S100A9 gene expression by WAY-169916 from spleens from the rat adjuvant model were confirmed by real-time RT–PCR (grey bars) compared with the regulation observed in the gene-profiling experiments (black bars). Results are expressed as means ± SEM from six rats per group. *P < 0.05 compared with vehicle control. (b) Treatment with WAY-169916 does not interfere with the binding of NF-κB to DNA. Nuclear extracts from primary mouse spleen cell cultures were co-treated for 18 hours with vehicle or concanavalin A (ConA; 5 μg/ml) and either WAY-169916 (1 μM) or pyrrolidine dithiocarbamate (PDTC) (100 μM) as indicated. CFA, complete Freund's adjuvant.
In addition, we attempted to determine whether treatment with WAY-169916 resulted in direct interference of NF-κB DNA binding in primary spleen cell cultures. The cells were stimulated with ConA (5 μg/ml) for 24 hours and co-treated with either WAY-169916 (1 μM) or PDTC (100 μM), a general inhibitor of NF-κB. As shown in Fig. 4b, activation by ConA resulted in an 80% increase in NF-κB DNA binding. Although PDTC treatment could completely block NF-κB activation, WAY-169916 was without effect. Control experiments demonstrated that the binding of NF-κB was specific, because competition experiments with wild-type oligonucleotide interfered with binding activity whereas a mutated oligonucleotide was without effect (data not shown). These results are consistent with our previous observations [16, 27] demonstrating that liganded ER inhibits NF-κB at the transcriptional level downstream from NF-κB DNA binding. Overall, these data indicate a marked anti-inflammatory effect for WAY-169916 that seems to cross multiple signaling pathways and tissues consistent with NF-κB's ubiquitous role in inflammation.
WAY-169916 anti-inflammatory activity in synoviocytes isolated from patients with RA
Finally, we wished to test whether WAY-169916 is active in FLS, a human cell type that is thought to have a pathologic function in joint destruction through its production of inflammatory cytokines and MMPs [32]. Activation of NF-κB in FLS is necessary for the production of these inflammatory mediators [5, 6]. FLS obtained from male patients with RA were stimulated with TNF-α and treated with vehicle, WAY-169916 (1 μM), or PDTC (100 μM). RNA was analyzed for gene expression changes of intercellular cell-adhesion molecule-1 (ICAM-1), IL-6, and TNF-α by real-time RT–PCR. As shown in Fig. 5, TNF-α-stimulated expression of all three inflammatory genes was significantly blocked by both WAY-169916 and PDTC, which was consistent with previous observations [5]. The cells were confirmed to express ERα mRNA [33] but no ERβ mRNA was detected (data not shown). In total, these data suggest the potential utility of non-steroidal selective NF-κB modulators such as WAY-169916 in treating patients with RA.
Inhibitory effect of treatment with WAY-169916 on inflammatory gene expression in FLS induced by TNF-α. Fibroblast-like synoviocytes (FLS) were pretreated for 1 hour with WAY-169916 (1 μM) or pyrrolidine dithiocarbamate (PDTC) (100 μM) before treatment with tumor necrosis factor-α (TNF-α) for 1 hour. The mRNA levels for TNF-α, IL-6 and intercellular cell-adhesion molecule-1 (ICAM-1) were determined by real-time RT–PCR and normalized to glyceraldehyde-3-phosphate dehydrogenase. Results are reported as means ± SEM for each group, with the mean level of the stimulated cells treated with vehicle defined as 1. *P < 0.05 compared with vehicle control.
RA might occur as a result of an autoimmune response, and recent studies suggest that hypersensitivity to microbial antigens contributes to the development of the arthritis. Microbial or self-antigen presentation to T lymphocytes results in chronic activation of the immune system. Multiple proinflammatory mediators, including IL-1, TNF-α, interferon-γ, and MMPs mediate the inflammation of the joints. Biochemical and histological changes in synovial tissue, cartilage, and bone have been documented in various animal models of arthritis [34, 35]. In many respects the synovial and cartilage lesions that develop in these models closely resemble those seen in rheumatoid arthritis. We have investigated the role of WAY-169916 in two such models.
The HLA-B27 transgenic rats spontaneously develop arthritis similar to the human spondyloarthropathies associated with the HLA-B27 and β2-microglobulin genes through a T cell-mediated process [34]. In this model, WAY-169916 restored the clinical joint scores to baseline after 10 days. Histological scoring of synovitis and cartilage damage in the tarsal joints after 29 days of treatment was also significantly improved with WAY-169916 treatment.
In the rat adjuvant-induced arthritis model [36], 3 to 6 days after the injection of adjuvant, induction of an αβ T cell response occurs and leads to clinical lesion development in the tarsal joints within 5 to 8 days. Because activated NF-κB was detected in the synovial lining layer and around blood vessels in the inflamed synovium as early as day 3 after adjuvant injection in the Lewis rats and is thought to be correlated with disease development [37], this model was used to test the therapeutic treatment with WAY-169916. We demonstrated that WAY-169916 was effective in improving both joint and histology scores at doses as low as 0.3 mg/kg given orally once daily. Improvement in the synovitis and Mankin scores did occur with higher doses of WAY-169916 even though the joint score reduction was already maximal at 0.3 mg/kg. When the rats were dosed at 5 mg/kg the total synovitis score decreased to 4.14 (data not shown). The beneficial effects of WAY-169916 on joint histology might therefore continue with increasing dose or longer exposure.
A benefit on arthritis progression with non-selective estrogens such a 17β-estradiol has also been demonstrated in both the rat adjuvant-induced arthritis model [38] and the collagen-induced mouse model [39, 40]. Indeed, 17β-estradiol has been shown to affect several processes involved in the pathogenesis of RA, including immunoregulation, regulation of adhesion molecules, and modulation of cytokine signaling. However, the role of estrogen has not been well defined in patients with RA. There is evidence that gender might affect the occurrence and progression of RA. Women have a higher risk of developing RA than men. During pregnancy, the disease activity is ameliorated in 75% of women, whereas after delivery, flares occur in up to 90% of patients [41]. The highest incidence of developing RA coincides with menopause, indicating that a decrease in estrogen production might increase the risk of joint inflammation. In a recent randomized clinical trial, post-menopausal women taking hormone therapy had suppressed signs of inflammation and significantly improved disease severity scores (DS28) after 12 months of treatment, which was consistent with previous trials [42].
With the identification of selective NF-κB transcriptional inhibitors such as WAY-169916, the expectation is to accentuate the anti-inflammatory, anti-rheumatic activity observed with the non-selective estrogens. Whereas non-selective estrogens have been documented to contain an anti-inflammatory activity through the suppression of NF-κB transcriptional activity [11, 12], hormone therapy can simultaneously elicit both proinflammatory and anti-inflammatory activities as exemplified by the decrease in haptoglobin and α1-AGP levels in women taking hormone therapy [43] while also inducing MMP9 and CRP levels [44, 45]. In the rat adjuvant model, WAY-169916 inhibited the adjuvant induction of CRP levels and those of haptoglobin and α1-AGP. Moreover, WAY-169916 had no effect on the basal levels of CRP whereas treatment with estradiol has been shown to increase rat CRP serum levels [46], suggesting a potential differential effect of WAY-169916 in comparison with estradiol. This differential activity has been demonstrated on several classic estrogenic effects. WAY-169916 neither stimulates creatine kinase gene expression driven via an estrogen receptor response element in vitro nor promotes uterine proliferation in vivo [27] while retaining the anti-inflammatory activity as demonstrated here.
The anti-inflammatory activity of WAY-169916 was further demonstrated in a series of gene-profiling experiments. In the spleens from the adjuvant-treated rats, 36 genes were identified that were induced greater than twofold by the adjuvant treatment. WAY-169916, when dosed at 5 mg/kg, repressed 29 of those genes by at least 50%, and 17 of them by more than 75%. An attractive hypothesis for WAY-169916-mediated activity in the spleen involves the downregulation of LBP and CD14 expression on monocytes and macrophages, resulting in a diminished immune response and ultimately resulting in the observed decreases in MMP9, IL1R2, chemokine-like factor 1, S100A8, and S100A9 through the repression of NF-κB activity [47, 48]. In spleen cell cultures, WAY-169916 treatment did not interfere with ConA-stimulated NF-κB DNA binding; however, the downregulation of S100A9 mRNA was confirmed (data not shown), which was consistent with our hypothesis that ER regulates NF-κB at the transcriptional level [16, 27]. A similar suppression of adjuvant-induced inflammatory gene expression was also observed in liver and lymph node studies. These data demonstrate that WAY-169916 can have an effect on a global level, both in terms of the tissues targeted and the different inflammatory signaling pathways, to suppress adjuvant-induced gene expression.
Infiltration of inflammatory cells into the synovial tissue and lining layer results in the formation of pannus, a highly vascularized tissue comprising FLS, macrophages, and lymphocytes. FLS are known for their role in joint destruction through the production of cytokines and MMP, which contribute to cartilage degradation (reviewed in [49]). Expression of ER has been detected in synovial tissues from patients with RA [50] and localized to synoviocytes in the synovial lining [22], providing another potential cell type by which WAY-169916 functions. Synoviocytes isolated from a male RA patient were confirmed to express ERα mRNA [33], but no ERβ mRNA was detected (data not shown). The ERα was functional in these cells, because WAY-169916 could effectively block the TNF-α-mediated inflammatory gene expression of IL-6, TNF-α, and ICAM-1. The potential involvement of NF-κB in mediating TNF-α gene induction was demonstrated with the use of a general NF-κB inhibitor, as shown previously [5]. Given the importance of TNF-α signaling in RA disease progression, the ability of WAY-169916 to interfere with this signaling pathway in human synoviocytes suggests a potential clinical benefit for WAY-169916 in patients with RA.
We detailed the activity of the first pathway-selective ER ligand, WAY-169916, in two models of RA. This compound selectively inhibits NF-κB activity via the ER and imparts significant efficacy in the HLA-B27 and Lewis rat adjuvant-induced models of arthritis. More importantly, no evidence for classic estrogenic activity has been observed with this compound [27]. These data provide evidence that the non-steroidal, pathway-selective ER ligand, WAY-169916, and other compounds in its class might be therapeutically useful in the treatment of RA.
α1-AGP:
α1-acid glycoprotein
ANOVA:
CFA:
complete Freund's adjuvant
ConA:
concanavalin A
CRP:
FLS:
fibroblast-like synoviocytes
ICAM-1:
intercellular cell-adhesion molecule-1
IκB:
inhibitory protein-κB
IL:
interleukin
LBP:
LBS binding protein
MMP:
matrix metalloproteinase
NF-κB:
nuclear factor-κB
PDTC:
pyrrolidine dithiocarbamate
RT–PCR:
TNF-α:
tumor necrosis factor-α.
Handel ML, McMorrow LB, Gravallese EM: Nuclear factor-kappa B in rheumatoid synovium. Localization of p50 and p65. Arthritis Rheum. 1995, 38: 1762-1770.
Asahara H, Asanuma M, Ogawa N, Nishibayashi S, Inoue H: High DNA-binding activity of transcription factor NF-kappa B in synovial membranes of patients with rheumatoid arthritis. Biochem Mol Biol Int. 1995, 37: 827-832.
Makarov SS: NF-kappa B in rheumatoid arthritis: a pivotal regulator of inflammation, hyperplasia, and tissue destruction. Arthritis Res. 2001, 3: 200-206. 10.1186/ar300.
Pahl HL: Activators and target genes of Rel/NF-kappaB transcription factors. Oncogene. 1999, 18: 6853-6866. 10.1038/sj.onc.1203239.
Fujisawa K, Aono H, Hasunuma T, Yamamoto K, Mita S, Nishioka K: Activation of transcription factor NF-kappa B in human synovial cells in response to tumor necrosis factor alpha. Arthritis Rheum. 1996, 39: 197-203.
Bondeson J, Foxwell B, Brennan F, Feldmann M: Defining therapeutic targets by using adenovirus: blocking NF-kappaB inhibits both inflammatory and destructive mechanisms in rheumatoid synovium but spares anti-inflammatory mediators. Proc Natl Acad Sci USA. 1999, 96: 5668-5673. 10.1073/pnas.96.10.5668.
Pope RM: Apoptosis as a therapeutic tool in rheumatoid arthritis. Nature Rev Immunol. 2002, 2: 527-535. 10.1038/nri846.
Seetharaman R, Mora AL, Nabozny G, Boothby M, Chen J: Essential role of T cell NF-kappa B activation in collagen-induced arthritis. J Immunol. 1999, 163: 1577-1583.
Campbell IK, Gerondakis S, O'Donnell K, Wicks IP: Distinct roles for the NF-kappaB1 (p50) and c-Rel transcription factors in inflammatory arthritis. J Clin Invest. 2000, 105: 1799-1806.
Tak PP, Gerlag DM, Aupperle KR, van de Geest DA, Overbeek M, Bennett BL, Boyle DL, Manning AM, Firestein GS: Inhibitor of nuclear factor kappaB kinase beta is a key regulator of synovial inflammation. Arthritis Rheum. 2001, 44: 1897-1907. 10.1002/1529-0131(200108)44:8<1897::AID-ART328>3.0.CO;2-4.
Stein B, Yang MX: Repression of the interleukin-6 promoter by estrogen receptor is mediated by NF-kappa B and C/EBP beta. Mol Cell Biol. 1995, 15: 4971-4979.
Ray A, Prefontaine KE, Ray P: Down-modulation of interleukin-6 gene expression by 17 beta-estradiol in the absence of high affinity DNA binding by the estrogen receptor. J Biol Chem. 1994, 269: 12940-12946.
Ray P, Ghosh SK, Zhang DH, Ray A: Repression of interleukin-6 gene expression by 17 beta-estradiol: inhibition of the DNA-binding activity of the transcription factors NF-IL6 and NF-kappa B by the estrogen receptor. FEBS Letters. 1997, 409: 79-85. 10.1016/S0014-5793(97)00487-0.
Deshpande R, Khalili H, Pergolizzi RG, Michael SD, Chang MD: Estradiol down-regulates LPS-induced cytokine production and NFκB activation in murine macrophages. Am J Reprod Immunol. 1997, 38: 46-54.
Sun WH, Keller ET, Stebler BS, Ershler WB: Estrogen inhibits phorbol ester-induced I kappa B alpha transcription and protein degradation. Biochem Biophys Res Commun. 1998, 244: 691-695. 10.1006/bbrc.1998.8324.
Harnish DC, Scicchitano MS, Adelman SJ, Lyttle CR, Karathanasis SK: The role of CBP in estrogen receptor cross-talk with nuclear factor-kappaB in HepG2 cells. Endocrinology. 2000, 141: 3403-3411. 10.1210/en.141.9.3403.
Speir E, Yu ZX, Takeda K, Ferrans VJ, Cannon RO: Competition for p300 regulates transcription by estrogen receptors and nuclear factor-kappa B in human coronary smooth muscle cells. Circ Res. 2000, 87: 1006-1011.
Kuiper GGJM, Carlsson B, Grandian K, Enmark E, Haggblad J, Nilsson S, Gustafsson JA: Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptor alpha and beta. Endocrinology. 1997, 138: 863-870. 10.1210/en.138.3.863.
Tornwall J, Carey AB, Fox RI, Fox HS: Estrogen in autoimmunity: expression of estrogen receptors in thymic and autoimmune T cells. J Gend Specif Med. 1999, 2: 33-40.
Ushiyama T, Ueyama H, Inoue K, Ohkubo I, Hukuda S: Expression of genes for estrogen receptors alpha and beta in human articular chondrocytes. Osteoartheritis Cartilage. 1999, 7: 560-566. 10.1053/joca.1999.0260.
Cutolo M, Accardo S, Villaggio B, Barone A, Sulli A, Coviello DA, Carabbio C, Felli L, Miceli D, Farruggio R, et al: Androgen and estrogen receptors are present in primary cultures of human synovial macrophages. J Clin Endocrinol Metab. 1996, 81: 820-827. 10.1210/jc.81.2.820.
Liu SH, al-Shaikh R, Panossian V, Yang RS, Nelson SD, Soleiman N, Finerman GA, Lane JM: Primary immunolocalization of estrogen and progesterone target cells in the human anterior cruciate ligament. J Orthop Res. 1996, 14: 526-533. 10.1002/jor.1100140405.
Cowley SM, Hoare S, Mosselman S, Parker MG: Estrogen receptors alpha and beta form heterodimers on DNA. J Biol Chem. 1997, 272: 19858-19862. 10.1074/jbc.272.32.19858.
Pace P, Taylor J, Suntharalingam S, Coombes RC, Ali S: Human estrogen receptor beta binds DNA in a manner similar to and dimerizes with estrogen receptor alpha. J Biol Chem. 1997, 272: 25832-25838. 10.1074/jbc.272.41.25832.
Tyree CM, Zou A, Allegretto EA: 17beta-Estradiol inhibits cytokine induction of the human E-selectin promoter. J Steroid Biochem Mol Biol. 2002, 80: 291-297. 10.1016/S0960-0760(02)00022-5.
Valentine JE, Kalkhoven E, White R, Hoare S, Parker MG: Mutations in the estrogen receptor ligand binding domain discriminate between hormone-dependent transactivation and transrepression. J Biol Chem. 2000, 275: 25322-25329. 10.1074/jbc.M002497200.
Chadwick CC, Chippari S, Matelan E, Borges-Marcucci L, Eckert AM, Keith JC, Albert LM, Leathurby Y, Harris HA, Bhat RA, et al: Identification of pathway selective estrogen receptor ligands that inhibit NF-κB transcriptional activity. Proc Natl Acad Sci USA.
Poole AR, Coombs RR: Rheumatoid-like joint lesions in rabbits injected intravenously with bovine serum. Int Arch Allergy Appl Immunol. 1977, 54: 97-113.
Mankin HJ, Dorfman H, Lippiello L, Zarins A: Biochemical and metabolic abnormalities in articular cartilage from osteo-arthritic human hips. II. Correlation of morphology with biochemical and metabolic data. J Bone Joint Surg. 1971, 53: 523-537.
Gianazza E, Eberini I, Villa P, Fratelli M, Pinna C, Wait R, Gemeiner M, Miller I: Monitoring the effects of drug treatment in rat models of disease by serum protein analysis. J Chromatogr B Analyt Technol Biomed Life Sci. 2002, 771: 107-130. 10.1016/S0378-4347(01)00562-X.
Otterness IG: The value of C-reactive protein measurement in rheumatoid arthritis. Semin Arthritis Rheum. 1994, 24: 91-104.
Muller-Ladner U, Gay RE, Gay S: Activation of synoviocytes. Curr Opin Rheumatol. 2000, 12: 186-194. 10.1097/00002281-200005000-00005.
Khalkhali-Ellis Z, Seftor EA, Nieva DR, Handa RJ, Price RH, Kirschmann DA, Baragi VM, Sharma RV, Bhalla RC, Moore TL, et al: Estrogen and progesterone regulation of human fibroblast-like synoviocyte function in vitro: implications in rheumatoid arthritis. J Rheum. 2000, 27: 1622-1631.
Taurog JD: Arthritis in HLA-B27 transgenic animals. Am J Med Sci. 1998, 316: 250-256. 10.1097/00000441-199810000-00005.
Waksman BH: Immune regulation in adjuvant disease and other arthritis models: relevance to pathogenesis of chronic arthritis. Scan J Immunol. 2002, 56: 12-34. 10.1046/j.1365-3083.2002.01106.x.
Van Eden W, Waksman BH: Immune regulation in adjuvant-induced arthritis: possible implications for innovative therapeutic strategies in arthritis. Arthritis Rheum. 2003, 48: 1788-1796. 10.1002/art.11077.
Tsao PW, Suzuki T, Totsuka R, Murata T, Takagi T, Ohmachi Y, Fujimura H, Takata I: The effect of dexamethasone on the expression of activated NF-kappa B in adjuvant arthritis. Clin Immunol Immunopathol. 1997, 83: 173-178. 10.1006/clin.1997.4333.
Badger AM, Blake SM, Dodds RA, Griswold DE, Swift BA, Rieman DJ, Stroup GB, Hoffman SJ, Gowen M: Idoxifene, a novel selective estrogen receptor modulator, is effective in a rat model of adjuvant-induced arthritis. J Pharmacol Exp Ther. 1999, 291: 1380-1386.
Jansson L, Olsson T, Holmdahl R: Estrogen induces a potent suppression of experimental autoimmune encephalomyelitis and collagen-induced arthritis in mice. J Neuroimmunol. 1994, 53: 203-207. 10.1016/0165-5728(94)90030-2.
Holmdahl R, Jansson L: Estrogen-induced suppression of collagen arthritis. III. Adult thymectomy does not affect the course of arthritis or the estrogen-mediated suppression of T-cell immunity. Brain Behav Immun. 1988, 2: 123-132. 10.1016/0889-1591(88)90013-X.
Kanik KS, Wilder RL: Hormonal alterations in rheumatoid arthritis, including the effects of pregnancy. Rheum Dis Clin North Am. 2000, 26: 805-823.
D'Elia HF, Larsen A, Mattsson LA, Waltbrand E, Kvist G, Mellstrom D, Saxne T, Ohlsson C, Nordborg E, Carlsten H: Influence of hormone replacement therapy on disease progression and bone mineral density in rheumatoid arthritis. J Rheum. 2003, 30: 1456-1463.
Tuck CH, Holleran S, Berglund L: Hormonal regulation of lipoprotein(a) levels: effects of estrogen replacement therapy on lipoprotein(a) and acute phase reactants in postmenopausal women. Arterioscler Thromb Vasc Biol. 1997, 17: 1822-1829.
Zanger D, Yang BK, Ardans J, Waclawiw MA, Csako G, Wahl LM, Cannon RO: Divergent effects of hormone therapy on serum markers of inflammation in postmenopausal women with coronary artery disease on appropriate medical management. J Am Coll Cardiol. 2000, 36: 1797-1802. 10.1016/S0735-1097(00)00952-9.
Pradhan AD, Manson JE, Rossouw JE, Siscovick DS, Mouton CP, Rifai N, Wallace RB, Jackson RD, Pettinger MB, Ridker PM: Inflammatory biomarkers, hormone replacement therapy, and incident coronary heart disease: prospective analysis from the Women's Health Initiative observational study. JAMA. 2002, 288: 980-987. 10.1001/jama.288.8.980.
El-Swefy SE, Asker ME, Ali SI, Mohammed HE: A novel concept to preserve the beneficial effects of hormone replacement therapy in bilaterally female ovariectomized rats: role of lovastatin therapy. Pharmacol Res. 2002, 45: 167-173. 10.1006/phrs.2001.0876.
Kido J, Kido R, Suryono , Kataoka M, Fagerhol MK, Nagata T: Calprotectin release from human neutrophils is induced by Porphyromonas gingivalis lipopolysaccharide via the CD-14–Toll-like receptor–nuclear factor kappaB pathway. J Periodontal Res. 2003, 38: 557-563. 10.1034/j.1600-0765.2003.00691.x.
Han W, Lou Y, Tang J, Zhang Y, Chen Y, Li Y, Gu W, Huang J, Gui L, Tang Y, et al: Molecular cloning and characterization of chemokine-like factor 1 (CKLF1), a novel human cytokine with unique structure and potential chemotactic activity. Biochem J. 2001, 357: 127-135. 10.1042/0264-6021:3570127.
McInnes IB, Leung BP, Liew FY: Cell–cell interactions in synovitis. Interactions between T lymphocytes and synovial cells. Arthritis Res. 2000, 2: 374-378. 10.1186/ar115.
Ushiyama T, Inoue K, Nishioka J: Expression of estrogen receptor related protein (p29) and estradiol binding in human arthritic synovium. J Rheum. 1995, 22: 421-426.
We thank all the members of the Discovery and Development Teams that contributed to this program.
Cardiovascular and Metabolic Disease Research, Wyeth Research, Cambridge, MA, USA
James C Keith Jr, Leo M Albert & Yelena Leathurby
Department Biological Technologies, Cambridge, MA, USA
Max Follettie & Lili Wang
Cardiovascular and Metabolic Disease Research, Collegeville, PA, USA
Lisa Borges-Marcucci & Douglas C Harnish
Women's Health Research Institute, Collegeville, PA, USA
Christopher C Chadwick
Chemical and Screening Sciences, Collegeville, PA, USA
Robert J Steffan
James C Keith Jr
Leo M Albert
Yelena Leathurby
Max Follettie
Lili Wang
Lisa Borges-Marcucci
Douglas C Harnish
Correspondence to Douglas C Harnish.
The authors are employees of Wyeth.
JCK, LMA and YL performed the in vivo experiments. MF and LW performed the gene-profiling experiments. LBM performed the cell-based assays and serum analysis. CCC, RJS and DCH were involved in the conception and identification of the molecule, and DCH wrote the manuscript. All authors contributed intellectually to the work and read and approved the final manuscript.
Keith, J.C., Albert, L.M., Leathurby, Y. et al. The utility of pathway selective estrogen receptor ligands that inhibit nuclear factor-κB transcriptional activity in models of rheumatoid arthritis. Arthritis Res Ther 7, R427 (2005). https://doi.org/10.1186/ar1692
Accepted: 17 January 2005
Popliteal Lymph Node
Inflammatory Gene Expression
Tarsal Joint
Adjuvant Induction


text:
My Developing Baby
Donating Cord Blood: The Gift of Life
When Joseph Krupski was gravely ill with a rare bone-marrow disease, another baby's banked cord blood saved him. Could you help another child?
By By Kate Ledger
Mary Krupski clearly remembers the doctor's voice on the other end of the phone saying, "I don't have anything good to tell you." She and her husband, Brian, had noticed that their then 2?-year-old son, Joseph, bruised easily, and he'd had a series of tests a few weeks earlier when he had a seemingly innocuous bump removed from his elbow. "We didn't even know that any more test results were coming," she says.
But a test revealed that Joseph had a rare disease called myelodysplastic syndrome (MDS), which was preventing the stem cells in his bone marrow from producing red and white blood cells. The Krupskis had
never heard of MDS, which strikes between 10,000 and 15,000 people annually, mostly adults. Redheaded, rambunctious Joseph didn't even look sick. Meeting with doctors at Children's Hospital Boston, three hours from their home, in Essex, Vermont, the Krupskis learned how aggressive Joseph's MDS was. "They said
the only cure would be a bone-marrow transplant to replace his abnormal stem cells," Mary recalls, "and
that without it he would probably die within two years."
Bone-marrow transplants, which have been performed for 40 years, require an exceptionally close genetic match between the patient and donor. Otherwise, the immune cells being introduced recognize the patient's body as "foreign" and reject it. The best matches for a bone-marrow transplant are usually siblings, but only 25 percent of patients have a brother or sister with a compatible tissue type. Joseph's two sisters, Allie and
Katey, then 8 and 6, were tested but weren't a match for him, and neither were his parents. The hospital launched a needle-in-a-haystack search to find a donor through the National Marrow Donor Program
(NMDP)'s Be The Match Registry of 8 million people who have volunteered to give bone marrow.
Mary, who had once worked as a clinical-trials coordinator at a breast-cancer center, and Brian, a medical-software developer, began reading online about the latest research that was happening at pediatric transplant centers. That's when they learned more about recent advances with umbilical-cord blood, an alternative
source of stem cells that was being used at a few other hospitals.
When Joseph was born, the Krupskis hadn't paid to store his own cord blood with a private bank. But even
if they had, the stored cells would have contained the same genetic defects of his disease -- and would have needed to be discarded. In fact, most children with a blood disease can't be treated with their own cells, according to the American Academy of Pediatrics, which recommends against private cord-blood storage except for newborns who have a sibling with a condition that could benefit from cord-blood transplantation.
Thankfully, there are also public cord-blood banks, which store potentially lifesaving stem cells that have
been donated by parents. "Cord blood doesn't need to be matched as closely to a patient as bone marrow
does," explains John Wagner, M.D., director of the division of hematology-oncology and blood and marrow transplantation at the University of Minnesota Amplatz Children's Hospital, in Minneapolis. "There's something unique about the immune cells in the placenta and the baby's umbilical-cord blood that reduces
the risk of graft-versus-host disease, the complication that results when the donor cells reject the patient,"
says Dr. Wagner, who helped pioneer the use of cord blood in the 1990s.
The Krupskis read about his work and flew to Minneapolis to meet him. What moved them most was the fact that cord blood was available immediately because it was already in the bank -- whereas it could take months for a bone-marrow donor to be found and then have a marrow-extraction procedure. They went home and packed up their entire family in March 2006 to move to Minnesota for the duration of Joseph's treatment.
The Cord Blood Controversy
Readers Weigh In
A Day In The Life of Liam: Pediatric Cancer Patient
Precious Resources
Public cord-blood banks first began flourishing in 2005, when President Bush signed the Stem Cell
Therapeutic and Research Act. The legislation restricted research on embryonic stem cells (which come
from a frozen embryo that ultimately gets destroyed) but also devoted $79 million to expand the nation's
cord-blood reserve for transplants and research. When parents decide to donate their baby's cord blood, a trained technician extracts about 3 teaspoons of blood minutes after the cord has been cut. Later, the blood
is tested for various infectious and hereditary diseases. Today, there are 19 public banks in the country
storing 160,000 units of cord blood at -196?C, which are available for anyone in the world who needs a transplant.
Cord blood is used primarily to treat children because one unit sometimes doesn't contain enough stem cells to be effective in an adult's larger body. Nearly 2,400 children have undergone cord-blood transplants so far,
500 of them last year. (In comparison, the NMDP was able to facilitate more than 4,000 marrow transplants
in 2010 in children and adults.)
Even though a transplant could save Joseph, his parents knew that the entire treatment process was very
risky. He first received high-dose chemotherapy and full-body radiation to destroy all the abnormal cells in
his body. Because this wiped out his immune system, he had to spend nearly four weeks in a hospital room
with purified air. "We left our shoes at the door, and wiped everything we could to avoid spreading germs," Mary recalls. She and Brian alternated sitting with Joseph, reading books and playing on a laptop with him,
and taking care of the girls at the nearby Ronald McDonald House, where they also went to school.
Finally, Joseph received the cord-blood transplant through an IV into a large blood vessel in his chest. The
stem cells miraculously travel through the bloodstream to reach the bone marrow, where they begin
producing healthy blood and immune cells. Still, the first 100 days after a transplant are critical. Vulnerable
to reactions and infections, Joseph stayed in the hospital for three weeks, and then he was allowed to live at
the Ronald McDonald House with his family (wearing a face mask at all times). The first biopsy test of his
bone marrow showed that 100 percent of the cells were the healthy donor cells, but a month later 10 percent of the cells were his own. Then he developed a high fever and a rash, signs of major complications including
graft-versus-host disease -- and a recurrence of his illness.
Stored Hope
This was a devastating blow. His MDS had now advanced and evolved into a form of leukemia. Based on
the literature that had been published on the condition at the time, Dr. Wagner says, Joseph's odds of
survival were zero. Mary had met other parents who'd lost their children and regretted that they hadn't
talked with them about death. So she sat with Joseph and held his hand. "I told him that God might have a
plan that he would go before us and be in Heaven with Jesus, and we would see him later, and that would be okay because he would be loved and he would be healthy. I didn't know if he understood because he was so sick and so young."
Joseph's only hope was a new strategy that Dr. Wagner had recently developed. Because he was so fragile, Joseph would have a less intense round of chemotherapy and radiation followed by a double cord-blood transplant -- meaning he would get two units of cord blood from two different babies. Having more stem
cells from two donors would offer a better opportunity for one type to thrive and produce new cells.
He got the second transplants in September, and the 100 days of waiting had to start all over again. Joseph stayed on a ventilator for a week, and because it prevented him from talking, he used hand signals to communicate with his parents. "We were amazed that he wasn't freaking out," Mary says. At one point, his fever spiked to 105?F, and his blood pressure and heart rate dropped alarmingly, but after three months he
was stable enough to return to the Ronald McDonald House.
The first biopsy showed that his marrow contained 50 percent of cells from each donor, but the next month
one donor type had taken hold and increased to 90 percent. Two months later, Joseph's marrow contained
100 percent donor cells -- which meant that the transplant was a success. Ten months after his first treatment had begun, he was healthy enough to go back to Vermont. "We were all home for Christmas, and it was
incredible," Mary remembers.
Four years later, Joseph is healthy and about to start third grade. Although the doctors prefer to say he's in remission because there's always a possibility that his disease could come back, "to us, he's cured," Mary
says. All the time he spent in isolation at the hospital -- as well as the following year when he had to stay
home to avoid germs -- has made him especially enthusiastic about playing with friends. "He'd be with
them around the clock if he could," says Mary.
The Krupskis keep a photo album of Joseph's treatment, and one of the pages has the labels from the cord-
blood units he received. Because of donor-confidentiality rules, all they know is that the stem cells that took hold came from a baby boy who would be about 9 years old now. "We're so thankful to any parents who
donate their baby's cord blood," Mary says. "Joseph would have died if he didn't have a transplant within a short period of time that second time around. We still feel as grateful for his life today as we were on the
day he was released from the hospital."
How to Donate Cord Blood
New awareness campaigns are encouraging parents to donate these potentially lifesaving cells, and there is a particular need for donations by parents of diverse ethnic and racial backgrounds. The National Marrow Donor Program's goal is to add an additional 150,000 units to the national inventory.
Parents have only been able to donate their baby's cord blood to a public bank if they deliver at one of about 200 hospitals that have an affiliation with a local public bank. (To see if your hospital is on the list, go to bethematch.org/cord. You must contact your doctor before your 34th week of pregnancy.) However, a new
pilot program -- at Duke University, in Durham, North Carolina; the M.D. Anderson Cancer Center, in
Houston; and the Texas Cord Blood Bank, in San Antonio -- is providing a special free mail-in kit for
parents anywhere who want to work with their doctor to donate their baby's cord blood. For information,
call 1-800-627-7692 at least six weeks before your baby is due.
In addition, through Cord Blood Registry's Designated Treatment Program, families who have an older child with a life-threatening disease that is treatable by donor cells can store their newborn's cord blood for free in a private bank. Go to cordblood.com for more information.
Originally published in the August 2011 issue of Parents magazine.
All content on this Web site, including medical opinion and any other health-related information, is for informational purposes only and should not be considered to be a specific diagnosis or treatment plan for any individual situation. Use of this site and the information contained herein does not create a doctor-patient relationship. Always seek the direct advice of your own doctor in connection with any questions or issues you may have regarding your own health or the health of others.
Popular in Cord Blood Banking
Labor Positions to Ease Pain (With Photos)
How Much Does Cord Blood Banking Cost?
Everything You Need to Know About First Trimester Prenatal Tests
The Perfect Labor and Delivery Nurse Gift
Postpartum Recovery: How to Make Life Easier (Really!)
Your Baby's Cord Blood
11 Valentine's Day Gifts Pregnant Women Really Want
Comparing Cord Blood Banking Companies
'Miracle' Newborn Whose Mom Died During Childbirth Will Be Taken Off Life Support, Dad Says
What You Need to Know About Cord Blood Banking
My Postpartum Tummy Tuck Was Life Changing
I Was a Traditional Surrogate and This is What it Was Like
The New Year's Baby Race: Could You Have a Contender?
If You're Overwhelmed by Cord Blood Banking Information, Read This
Pregnancy Complication: Differences in the Umbilical Cord and Placenta
Borderline High Blood Pressure and Pregnancy
Is Anal Sex Safe During Pregnancy?
Should You Bank Your Baby's Cord Blood?
My Aunt Announced My Pregnancy on Facebook Without My Consent
I Lost So Much Weight During Pregnancy Due to Hyperemesis Gravidarum
All Topics in Cord Blood Banking
Twins and Multiples


Baraboo Area Literacy Council – Want to work on reading skills? We can help!
Want to work on reading skills? We can help!
Need help with reading and writing?
Free adult tutoring is available.
Contact your library – Baraboo Public Library, 230 Fourth Avenue 608-356-6166 or stop in to fill out a Tutor Request form.
The Baraboo Area Literacy Council (BALC) was established in 1986 to address literacy needs in Baraboo and the surrounding communities. The organization is volunteer and non-profit.
BALC provides tutors for adults who need help with basic literacy (reading and writing), or English as a second language (ESL). Tutors and students meet on a one-to-one basis, in a safe place, at their mutual convenience. The service is free and continuous. BALC tutors are trained volunteers who have agreed to a background check.
The BALC welcomes community support and is always looking for new members. The BALC – its board, its tutor coordinator and all tutors – is made up entirely of volunteers. If interested in joining the BALC, either as a tutor or as a board member, please leave your name and contact information, including email if available, at the circulation desk of the Baraboo Public Library or email boolit@barabooliteracy.org.
If you don't have time or interest in participating actively in the BALC program, monetary contributions are welcome and needed. Since we are a non-profit group your donation would be tax-deductible. Checks may be made out to BALC and left at the desk of the Baraboo Public Library.


text:
#DiscoverON Festivals & Events
Halloween Haunts In Ontario. Warning: They're Not Just For Kids.
Halloween is a highlight each October for kids who love costumes, trick-or-treating and gorging on chocolate and candy. But in Ontario, you'll find plenty of ghoulishness for the grown-up kids-at-heart among us who'll never outgrow the need for fright-filled fun.
From horror film festivals, weird and sinister walking tours, scare parks, eerie hotels and deserted ghost towns, here are five spooktacular Ontario cities you'll need to haunt this Halloween season.
Spend a few days delving into the dark side of Canada's original capital.
Kingston's original haunted walk through old Sydenham Ward features haunted hotels, tales of hidden burial grounds and grave robbings, the site of hangings at the old courthouse, and the city's famous haunted courtyard. Or ride the Ghost and Mystery Trolley Tour through Kingston's paranormal past to haunted spots like the Rockwood Asylum, the Cataraqui Cemetery and the grave of Sir John A. Macdonald.
Visit the infamous Kingston Penitentiary that once confined Canada's most notorious criminals. Enter cells, segregation rooms and recreation yards on the 90-minute guided tour and expect full body goosebumps.
Fort Henry National Historic Site is a UNESCO World Heritage 19th century military living museum that transforms into one of North America's top haunted houses each Halloween season. Fort Fright taps into our deepest fears with nightmarish scenes of death, decay and creepy clowns.
Don't be surprised if you rub shoulders with a supernatural guest at the Hochelaga Inn, a beautiful 1879 Victorian mansion and one of Kingston's finest places to sleep (with one eye open).
Northern Ontario's largest city grew from its humble roots as a mining town and holds a myriad of dark tales from its struggle to prosperity.
Due to the rise and fall of the mining industry, there are many defunct mine pits and ghost towns surrounding Sudbury, like Sellwood, abandoned in 1932. Today, if you take a walk where the town once stood, all you'll find is a scattering of ruins and the graves of three young children.
Check out the special Halloween themed frights and delights taking place at Science North and Dynamic Earth, including the Tunnel of Terror. Do you dare?
Just south of Sudbury is the site of Burwash Camp Bison, an abandoned prison that now sits on a private farm. Contact the property owners directly for permission to visit and to arrange a tour of the place.
Catch the annual terror train to Sellwood Asylum at the Northern Ontario Railroad Museum and Heritage Center, in Capreol, in the north end of the city along the Vermilion River. And get your scare on at the 5th Northern Frights Festival, Northern Ontario's premiere horror film festival taking place over the course of one nightmare-filled day on October 19 at the Sudbury Theatre Centre.
Our nation's capital comes in at a solid 10 out of 10 on the scare scale.
The Haunted Walk hosts several spinetingling tours. Descend into the city's paranormal past on the original haunted walk, tours of haunted jails and the homes of former Prime Ministers, or embark on an interactive zombie adventure deep inside the Diefenbunker Museum, a former military bunker from the Cold War era.
Tucked away beside the Bytown Locks, the Bytown Museum, is one of the city's oldest buildings and is said to be home to the restless spirit of its construction manager.
The HI Ottawa Jail Hostel on Nicholas Street is an old jailhouse turned accommodation for the budget conscious. Believed to be the place where an assassin was hanged in 1869, guests have reported seeing his apparition and those of other prisoners staring back at them. Take the free daily jail tour through death row, have a drink at the hostel's bar, Mugshots, and if you dare, sleep (or try to) in a real solitary confinement cell. Alternatively, check into the grand, historic Hotel Fairmont Château Laurier and keep an eye out for the phantom of its previous owner, who tragically perished on the Titanic.
Canada's most cosmopolitan city has its fair share of scare.
Facing Toronto's harbourfront are a cluster of scenic islands, popular among daytrippers during the summer but much quieter during the rest of the year. On Centre Island, hike out to the Gibraltar Point Lighthouse, the oldest standing lighthouse on the Great Lakes, built in the early 1800s. Its first lightkeeper was also a bootlegger and is believed to haunt the spot of his rumored murder by drunken soldiers.
Casa Loma, the fabled gothic mansion atop the city, opens its doors this season for Legends of Horror, an hour-long, self-guided sleuth through the mansion past nooks and crannies not usually open to the public.
For horror film fans, Toronto After Dark presents nine days of the best new thrills and chills from the world of cinema, October 17-25. While in town, you'll also want to check out the 6ix Screams International Horror Short Film Festival on October 21. And yet another must-see for fans of the genre is the It's Alive! exhibit at the Royal Ontario Museum, showcasing classic horror and sci-fi art from the private collection of Kirk Hammett, the legendary guitarist of heavy metal band Metallica.
For live theatre fans, a visit to the historic Elgin & Winter Garden Theatres is a must. Catch the smash hit Come From Away, and on October 28, take the rarely offered Ghost Tour. A lonely trombone player and a lady who prefers lavender perfume are said to haunt the national historic site that dates back to 1913 and the height of Vaudeville.
According to legend, screams and footsteps heard in the 19th floor stairwell of The Fairmont Royal York, one of Toronto's most distinguished historic hotels, belong to a former employee who mysteriously died there. Guests and staff have also reported the service elevator riding up to the closed Crystal Ballroom without anyone having pushed the floor button.
Image credit: Fear Farm
KITCHENER/WATERLOO
Just an hour outside of Toronto are the twin cities of Kitchener and Waterloo. You're in Fear Farm country now.
Experience bone chilling terror at Bingemans Screampark for four horrifying haunts, three execution rides and one red escape room. About half an hour drive west in Bright is Fear Farm, Ontario's legendary haunted farm, not for the faint of heart. Snyder's Farm is transformed into a horror park with six haunt experiences, wandering zombies, fire eaters, illusionists, fortune tellers and a haunted house.
If you survive, take a wander over to the Waterloo County Jail and the Governor's House in Kitchener. Built in 1852, the jail is the oldest existing government building in the city and is said to be haunted by spirits of former inmates. While in town, experience the historic charm of The Walper Hotel, an iconic 1893 downtown landmark boutique hotel, reputed to be inhabited by a benevolent paranormal presence.
And just a 45-minute drive west, in Ontario's renowned theatre town of Stratford, the guided Ghosts of Stratford Theatrical Walking Tours invite you to drown your fears on the Pubs, Pilsners and Spirits walk or take in one more nightmare on the special Halloween edition Dr. Blood Walks Again tour.
If you experience any of these or other ghoulish gems in Ontario, snap a pic and share it. We're dying to hear all about it. Happy Halloween!
← 8 Foodie Destinations In Ontario You Need To Savour This Fall → Fall Road Trip: Twenty Valley to Niagara Falls


If you need to write on leather to demarcate an area in need of repair, you should use leather writing pens rather than fabric markers. But not all leather markers are made alike. Learn why LeatherPen brand markers are the ideal choice for your hide projects.
Unlike regular pens, which are so thin they tend to give your hand cramps, LeatherPen markers are made slightly thicker. This gives you a good, solid grip on the pen, allowing for precise marking and cramp-free writing.
You can extend the life of your marker pen by replacing the cap snugly after each use. The caps on these leather writing pens snap on and off easily, allowing for quick removal when needed. Thus, you don't have to feel as though you need to leave the caps off for fast access to the ink.
Some markers require special priming techniques before finally flowing out onto the cloth. These leather writing pens only need a few sharp taps before flowing effortlessly onto the hide. Once the flow is started, you can start and stop your drawing as necessary. The pen will pick up seamlessly where you left off and continue a steady stream of ink until you finish drawing around the flaw.
No one wants to buy pack after pack of leather writing pens just because they dried out or were used up quickly. With LeatherPen markers, you get the maximum amount of life out of each pen. Draw with the nib in its native position until you can no longer coax any more ink out of the tip. Then, pop the nib out, flip it around and put it back in. The other side will give you several more uses before you need to replace the nib. To replace it, pop it out as you did to reverse it, and then insert the new nib in its place. It's that simple!
One pen color doesn't match all hides. In fact, you don't want it to match at all. The whole objective in using leather writing pens is to make each flaw more visible. That's why LeatherPen offers markers in blue, green, brown and white. You never have to worry about the marker staining the leather, either. After the markings have served their purpose, take a dry cloth to wipe off the ink.


Whether it's heart-shaped, '80s-song inspired or pepperoni flecked, Aldi loves cheese (and, same).
So when we heard the neighborhood's favorite grocery store was releasing a St. Patrick's Day collection of the dairy goodness, we were all ears (and tastebuds).
The new collection will feature five types of Happy Farms cheese: Pesto Gouda, English Sage Derby, Aged Irish Cheddar, Irish Cheddar with Beer and Irish Cheddar with Whiskey. Yes, two of the cheeses come infused with alcohol, so we're feeling as lucky as a four-leaf clover.
The cheese will be in stores beginning February 27, and two are a delightful shade of Kelly green, meaning they're the ideal excuse to host a St. Patrick's Day party (even if it's just a party of one) this March 17.
The best part: Each cheese is $4.
So, yeah, you don't need a pot full o' gold to get your hands on these pressed milk curds.


Fun Comes in Many Sizes.
One look at the Metropolitan and you just can't help but think "fun." Fun to ride, fun to look at (and be seen on!), and, thanks to its gas-sipping thriftiness and incredible pricing, even more fun to own.
Meet the bad boy of Honda's scooter family, the urban, street-smart hustler that packs as much feisty, fuel-efficient performance as it does gritty attitude. Want to make a statement? Then ride a Ruckus.
Looking for a sharp scooter that can do it all, from urban runabout to a longer ride? Hook up with a Honda PCX150. It's got to be one of the best transportation values anywhere, it's easy to operate, and best of all it's a blast to ride.
The Honda Forza is one of the most practical scooters anywhere. Its 300-class engine has plenty of power—even two-up—and gets great mileage. Underseat storage makes errands a breeze. And the sleek bodywork means you'll arrive in style.