prompt_front: Now, answer the following question based on the text below, only give me the answer and do not output any other words.

Question: What do dendritic spines contain?

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Context 1: JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
A role for thrombospondin-1 deficits in astrocyte-mediated spine and synaptic pathology in Downs syndrome  Octavio Garcia, Maria Torres, Pablo Helguera, Pinar Coskun, Jorge Busciglio 
PUBLISHED: 07-02-2010	Downs syndrome (DS) is the most common genetic cause of mental retardation  Reduced number and aberrant architecture of dendritic spines are common features of DS neuropathology  However, the mechanisms involved in DS spine alterations are not known  In addition to a relevant role in synapse formation and maintenance, astrocytes can regulate spine dynamics by releasing soluble factors or by physical contact with neurons  We have previously shown impaired mitochondrial function in DS astrocytes leading to metabolic alterations in protein processing and secretion  In this study, we investigated whether deficits in astrocyte function contribute to DS spine pathology 
Analysis of Dendritic Spine Morphology in Cultured CNS Neurons Authors: Deepak P  Srivastava, Kevin M  Woolfrey, Peter Penzes  Published: 07-13-2011 JoVE Neuroscience
Dendritic spines are the sites of the majority of excitatory connections within the brain, and form the post-synaptic compartment of synapses  These structures are rich in actin and have been shown to be highly dynamic  In response to classical Hebbian plasticity as well as neuromodulatory signals, dendritic spines can change shape and number, which is thought to be critical for the refinement of neural circuits and the processing and storage of information within the brain  Within dendritic spines, a complex network of proteins link extracellular signals with the actin cyctoskeleton allowing for control of dendritic spine morphology and number  Neuropathological studies have demonstrated that a number of disease states, ranging from schizophrenia to autism spectrum disorders, display abnormal dendritic spine morphology or numbers  Moreover, recent genetic studies have identified mutations in numerous genes that encode synaptic proteins, leading to suggestions that these proteins may contribute to aberrant spine plasticity that, in part, underlie the pathophysiology of these disorders  In order to study the potential role of these proteins in controlling dendritic spine morphologies/number, the use of cultured cortical neurons offers several advantages  Firstly, this system allows for high-resolution imaging of dendritic spines in fixed cells as well as time-lapse imaging of live cells  Secondly, this in vitro system allows for easy manipulation of protein function by expression of mutant proteins, knockdown by shRNA constructs, or pharmacological treatments  These techniques allow researchers to begin to dissect the role of disease-associated proteins and to predict how mutations of these proteins may function in vivo 
Play ButtonIsolation and Culture of Mouse Cortical AstrocytesAuthors: Sebastian Schildge, Christian Bohrer, Kristina Beck, Christian Schachtrup  Institutions: University of Freiburg , University of Freiburg  Astrocytes are an abundant cell type in the mammalian brain, yet much remains to be learned about their molecular and functional characteristics  In vitro astrocyte cell culture systems can be used to study the biological functions of these glial cells in detail  This video protocol shows how to obtain pure astrocytes by isolation and culture of mixed cortical cells of mouse pups  The method is based on the absence of viable neurons and the separation of astrocytes, oligodendrocytes and microglia, the three main glial cell populations of the central nervous system, in culture  Representative images during the first days of culture demonstrate the presence of a mixed cell population and indicate the timepoint, when astrocytes become confluent and should be separated from microglia and oligodendrocytes  Moreover, we demonstrate purity and astrocytic morphology of cultured astrocytes using immunocytochemical stainings for well established and newly described astrocyte markers  This culture system can be easily used to obtain pure mouse astrocytes and astrocyte-conditioned medium for studying various aspects of astrocyte biology Neuroscience, Issue 71, Neurobiology, Cellular Biology, Medicine, Molecular Biology, Anatomy, Physiology, brain, mouse, astrocyte culture, astrocyte, fibroblast, fibrinogen, chondroitin sulfate proteoglycan, neuronal regeneration, cell culture, animal model50079Play ButtonImaging Dendritic Spines of Rat Primary Hippocampal Neurons using Structured Illumination MicroscopyAuthors: Marijn Schouten, Giulia M  R  De Luca, Diana K  Alatriste González, Babette E  de Jong, Wendy Timmermans, Hui Xiong, Harm Krugers, Erik M  M  Manders, Carlos P  Fitzsimons  Institutions: University of Amsterdam, University of Amsterdam Dendritic spines are protrusions emerging from the dendrite of a neuron and represent the primary postsynaptic targets of excitatory inputs in the brain  Technological advances have identified these structures as key elements in neuron connectivity and synaptic plasticity  The quantitative analysis of spine morphology using light microscopy remains an essential problem due to technical limitations associated with light's intrinsic refraction limit  Dendritic spines can be readily identified by confocal laser-scanning fluorescence microscopy  However, measuring subtle changes in the shape and size of spines is difficult because spine dimensions other than length are usually smaller than conventional optical resolution fixed by light microscopy's theoretical resolution limit of 200 nm 
Several recently developed super resolution techniques have been used to image cellular structures smaller than the 200 nm, including dendritic spines  These techniques are based on classical far-field operations and therefore allow the use of existing sample preparation methods and to image beyond the surface of a specimen  Described here is a working protocol to apply super resolution structured illumination microscopy (SIM) to the imaging of dendritic spines in primary hippocampal neuron cultures  Possible applications of SIM overlap with those of confocal microscopy  However, the two techniques present different applicability  SIM offers higher effective lateral resolution, while confocal microscopy, due to the usage of a physical pinhole, achieves resolution improvement at the expense of removal of out of focus light  In this protocol, primary neurons are cultured on glass coverslips using a standard protocol, transfected with DNA plasmids encoding fluorescent proteins and imaged using SIM  The whole protocol described herein takes approximately 2 weeks, because dendritic spines are imaged after 16-17 days in vitro, when dendritic development is optimal  After completion of the protocol, dendritic spines can be reconstructed in 3D from series of SIM image stacks using specialized software Neuroscience, Issue 87, Dendritic Spine, Microscopy, Confocal, Fluorescence, Neurosciences, hippocampus, primary neuron, super resolution microscopy, structured illumination microscopy (SIM), neuroscience, dendrite51276Play ButtonSetting-up an In Vitro Model of Rat Blood-brain Barrier (BBB): A Focus on BBB Impermeability and Receptor-mediated TransportAuthors: Yves Molino, Françoise Jabès, Emmanuelle Lacassagne, Nicolas Gaudin, Michel Khrestchatisky  Institutions: VECT-HORUS SAS, CNRS, NICN UMR 7259 The blood brain barrier (BBB) specifically regulates molecular and cellular flux between the blood and the nervous tissue  Our aim was to develop and characterize a highly reproducible rat syngeneic in vitro model of the BBB using co-cultures of primary rat brain endothelial cells (RBEC) and astrocytes to study receptors involved in transcytosis across the endothelial cell monolayer  Astrocytes were isolated by mechanical dissection following trypsin digestion and were frozen for later co-culture  RBEC were isolated from 5-week-old rat cortices  The brains were cleaned of meninges and white matter, and mechanically dissociated following enzymatic digestion  Thereafter, the tissue homogenate was centrifuged in bovine serum albumin to separate vessel fragments from nervous tissue  The vessel fragments underwent a second enzymatic digestion to free endothelial cells from their extracellular matrix  The remaining contaminating cells such as pericytes were further eliminated by plating the microvessel fragments in puromycin-containing medium  They were then passaged onto filters for co-culture with astrocytes grown on the bottom of the wells  RBEC expressed high levels of tight junction (TJ) proteins such as occludin, claudin-5 and ZO-1 with a typical localization at the cell borders  The transendothelial electrical resistance (TEER) of brain endothelial monolayers, indicating the tightness of TJs reached 300 ohm·cm2 on average  The endothelial permeability coefficients (Pe) for lucifer yellow (LY) was highly reproducible with an average of 0 26 ± 0 11 x 10-3 cm/min  Brain endothelial cells organized in monolayers expressed the efflux transporter P-glycoprotein (P-gp), showed a polarized transport of rhodamine 123, a ligand for P-gp, and showed specific transport of transferrin-Cy3 and DiILDL across the endothelial cell monolayer  In conclusion, we provide a protocol for setting up an in vitro BBB model that is highly reproducible due to the quality assurance methods, and that is suitable for research on BBB transporters and receptors Medicine, Issue 88, rat brain endothelial cells (RBEC), mouse, spinal cord, tight junction (TJ), receptor-mediated transport (RMT), low density lipoprotein (LDL), LDLR, transferrin, TfR, P-glycoprotein (P-gp), transendothelial electrical resistance (TEER),51278Play ButtonInducing Plasticity of Astrocytic Receptors by Manipulation of Neuronal Firing RatesAuthors: Alison X  Xie, Kelli Lauderdale, Thomas Murphy, Timothy L  Myers, Todd A  Fiacco  Institutions: University of California Riverside, University of California Riverside, University of California Riverside Close to two decades of research has established that astrocytes in situ and in vivo express numerous G protein-coupled receptors (GPCRs) that can be stimulated by neuronally-released transmitter  However, the ability of astrocytic receptors to exhibit plasticity in response to changes in neuronal activity has received little attention  Here we describe a model system that can be used to globally scale up or down astrocytic group I metabotropic glutamate receptors (mGluRs) in acute brain slices  Included are methods on how to prepare parasagittal hippocampal slices, construct chambers suitable for long-term slice incubation, bidirectionally manipulate neuronal action potential frequency, load astrocytes and astrocyte processes with fluorescent Ca2+ indicator, and measure changes in astrocytic Gq GPCR activity by recording spontaneous and evoked astrocyte Ca2+ events using confocal microscopy  In essence, a “calcium roadmap” is provided for how to measure plasticity of astrocytic Gq GPCRs  Applications of the technique for study of astrocytes are discussed  Having an understanding of how astrocytic receptor signaling is affected by changes in neuronal activity has important implications for both normal synaptic function as well as processes underlying neurological disorders and neurodegenerative disease Neuroscience, Issue 85, astrocyte, plasticity, mGluRs, neuronal Firing, electrophysiology, Gq GPCRs, Bolus-loading, calcium, microdomains, acute slices, Hippocampus, mouse51458Play ButtonInhibitory Synapse Formation in a Co-culture Model Incorporating GABAergic Medium Spiny Neurons and HEK293 Cells Stably Expressing GABAA ReceptorsAuthors: Laura E  Brown, Celine Fuchs, Martin W  Nicholson, F  Anne Stephenson, Alex M  Thomson, Jasmina N  Jovanovic  Institutions: University College London Inhibitory neurons act in the central nervous system to regulate the dynamics and spatio-temporal co-ordination of neuronal networks  GABA (γ-aminobutyric acid) is the predominant inhibitory neurotransmitter in the brain  It is released from the presynaptic terminals of inhibitory neurons within highly specialized intercellular junctions known as synapses, where it binds to GABAA receptors (GABAARs) present at the plasma membrane of the synapse-receiving, postsynaptic neurons  Activation of these GABA-gated ion channels leads to influx of chloride resulting in postsynaptic potential changes that decrease the probability that these neurons will generate action potentials  During development, diverse types of inhibitory neurons with distinct morphological, electrophysiological and neurochemical characteristics have the ability to recognize their target neurons and form synapses which incorporate specific GABAARs subtypes  This principle of selective innervation of neuronal targets raises the question as to how the appropriate synaptic partners identify each other  To elucidate the underlying molecular mechanisms, a novel in vitro co-culture model system was established, in which medium spiny GABAergic neurons, a highly homogenous population of neurons isolated from the embryonic striatum, were cultured with stably transfected HEK293 cell lines that express different GABAAR subtypes  Synapses form rapidly, efficiently and selectively in this system, and are easily accessible for quantification  Our results indicate that various GABAAR subtypes differ in their ability to promote synapse formation, suggesting that this reduced in vitro model system can be used to reproduce, at least in part, the in vivo conditions required for the recognition of the appropriate synaptic partners and formation of specific synapses  Here the protocols for culturing the medium spiny neurons and generating HEK293 cells lines expressing GABAARs are first described, followed by detailed instructions on how to combine these two cell types in co-culture and analyze the formation of synaptic contacts  Neuroscience, Issue 93, Developmental neuroscience, synaptogenesis, synaptic inhibition, co-culture, stable cell lines, GABAergic, medium spiny neurons, HEK 293 cell line52115Play ButtonTwo-Photon in vivo Imaging of Dendritic Spines in the Mouse Cortex Using a Thinned-skull PreparationAuthors: Xinzhu Yu, Yi Zuo  Institutions: University of California, Santa Cruz In the mammalian cortex, neurons form extremely complicated networks and exchange information at synapses  Changes in synaptic strength, as well as addition/removal of synapses, occur in an experience-dependent manner, providing the structural foundation of neuronal plasticity  As postsynaptic components of the most excitatory synapses in the cortex, dendritic spines are considered to be a good proxy of synapses  Taking advantages of mouse genetics and fluorescent labeling techniques, individual neurons and their synaptic structures can be labeled in the intact brain  Here we introduce a transcranial imaging protocol using two-photon laser scanning microscopy to follow fluorescently labeled postsynaptic dendritic spines over time in vivo  This protocol utilizes a thinned-skull preparation, which keeps the skull intact and avoids inflammatory effects caused by exposure of the meninges and the cortex  Therefore, images can be acquired immediately after surgery is performed  The experimental procedure can be performed repetitively over various time intervals ranging from hours to years  The application of this preparation can also be expanded to investigate different cortical regions and layers, as well as other cell types, under physiological and pathological conditions Neuroscience, Issue 87, dendritic spine, mouse cortex, in vivo, two-photon microscopy, thinned-skull, imaging51520Play ButtonModeling Astrocytoma Pathogenesis In Vitro and In Vivo Using Cortical Astrocytes or Neural Stem Cells from Conditional, Genetically Engineered MiceAuthors: Robert S  McNeill, Ralf S  Schmid, Ryan E  Bash, Mark Vitucci, Kristen K  White, Andrea M  Werneke, Brian H  Constance, Byron Huff, C  Ryan Miller  Institutions: University of North Carolina School of Medicine, University of North Carolina School of Medicine, University of North Carolina School of Medicine, University of North Carolina School of Medicine, University of North Carolina School of Medicine, Emory University School of Medicine, University of North Carolina School of Medicine Current astrocytoma models are limited in their ability to define the roles of oncogenic mutations in specific brain cell types during disease pathogenesis and their utility for preclinical drug development  In order to design a better model system for these applications, phenotypically wild-type cortical astrocytes and neural stem cells (NSC) from conditional, genetically engineered mice (GEM) that harbor various combinations of floxed oncogenic alleles were harvested and grown in culture  Genetic recombination was induced in vitro using adenoviral Cre-mediated recombination, resulting in expression of mutated oncogenes and deletion of tumor suppressor genes  The phenotypic consequences of these mutations were defined by measuring proliferation, transformation, and drug response in vitro  Orthotopic allograft models, whereby transformed cells are stereotactically injected into the brains of immune-competent, syngeneic littermates, were developed to define the role of oncogenic mutations and cell type on tumorigenesis in vivo  Unlike most established human glioblastoma cell line xenografts, injection of transformed GEM-derived cortical astrocytes into the brains of immune-competent littermates produced astrocytomas, including the most aggressive subtype, glioblastoma, that recapitulated the histopathological hallmarks of human astrocytomas, including diffuse invasion of normal brain parenchyma  Bioluminescence imaging of orthotopic allografts from transformed astrocytes engineered to express luciferase was utilized to monitor in vivo tumor growth over time  Thus, astrocytoma models using astrocytes and NSC harvested from GEM with conditional oncogenic alleles provide an integrated system to study the genetics and cell biology of astrocytoma pathogenesis in vitro and in vivo and may be useful in preclinical drug development for these devastating diseases Neuroscience, Issue 90, astrocytoma, cortical astrocytes, genetically engineered mice, glioblastoma, neural stem cells, orthotopic allograft51763Play ButtonPaired Whole Cell Recordings in Organotypic Hippocampal SlicesAuthors: Chantelle Fourie, Marianna Kiraly, Daniel V  Madison, Johanna M  Montgomery  Institutions: University of Auckland, Stanford University Pair recordings involve simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling not only direct electrophysiological characterization of the synaptic connections between individual neurons, but also pharmacological manipulation of either the presynaptic or the postsynaptic neuron  When carried out in organotypic hippocampal slice cultures, the probability that two neurons are synaptically connected is significantly increased  This preparation readily enables identification of cell types, and the neurons maintain their morphology and properties of synaptic function similar to that in native brain tissue  A major advantage of paired whole cell recordings is the highly precise information it can provide on the properties of synaptic transmission and plasticity that are not possible with other more crude techniques utilizing extracellular axonal stimulation  Paired whole cell recordings are often perceived as too challenging to perform  While there are challenging aspects to this technique, paired recordings can be performed by anyone trained in whole cell patch clamping provided specific hardware and methodological criteria are followed  The probability of attaining synaptically connected paired recordings significantly increases with healthy organotypic slices and stable micromanipulation allowing independent attainment of pre- and postsynaptic whole cell recordings  While CA3-CA3 pyramidal cell pairs are most widely used in the organotypic slice hippocampal preparation, this technique has also been successful in CA3-CA1 pairs and can be adapted to any neurons that are synaptically connected in the same slice preparation  In this manuscript we provide the detailed methodology and requirements for establishing this technique in any laboratory equipped for electrophysiology Neuroscience, Issue 91, hippocampus, paired recording, whole cell recording, organotypic slice, synapse, synaptic transmission, synaptic plasticity51958Play ButtonImaging Intracellular Ca2+ Signals in Striatal Astrocytes from Adult Mice Using Genetically-encoded Calcium IndicatorsAuthors: Ruotian Jiang, Martin D  Haustein, Michael V  Sofroniew, Baljit S  Khakh  Institutions: University of California Los Angeles, University of California Los Angeles Astrocytes display spontaneous intracellular Ca2+ concentration fluctuations ([Ca2+]i) and in several settings respond to neuronal excitation with enhanced [Ca2+]i signals  It has been proposed that astrocytes in turn regulate neurons and blood vessels through calcium-dependent mechanisms, such as the release of signaling molecules  However, [Ca2+]i imaging in entire astrocytes has only recently become feasible with genetically encoded calcium indicators (GECIs) such as the GCaMP series  The use of GECIs in astrocytes now provides opportunities to study astrocyte [Ca2+]i signals in detail within model microcircuits such as the striatum, which is the largest nucleus of the basal ganglia  In the present report, detailed surgical methods to express GECIs in astrocytes in vivo, and confocal imaging approaches to record [Ca2+]i signals in striatal astrocytes in situ, are described  We highlight precautions, necessary controls and tests to determine if GECI expression is selective for astrocytes and to evaluate signs of overt astrocyte reactivity  We also describe brain slice and imaging conditions in detail that permit reliable [Ca2+]i imaging in striatal astrocytes in situ  The use of these approaches revealed the entire territories of single striatal astrocytes and spontaneous [Ca2+]i signals within their somata, branches and branchlets  The further use and expansion of these approaches in the striatum will allow for the detailed study of astrocyte [Ca2+]i signals in the striatal microcircuitry Neuroscience, Issue 93, astrocyte, calcium, striatum, GECI, GCaMP3, AAV2/5, stereotaxic injection, brain slice, imaging51972Play ButtonMethods to Assess Subcellular Compartments of Muscle in C  elegansAuthors: Christopher J  Gaffney, Joseph J  Bass, Thomas F  Barratt, Nathaniel J  Szewczyk  Institutions: University of Nottingham Muscle is a dynamic tissue that responds to changes in nutrition, exercise, and disease state  The loss of muscle mass and function with disease and age are significant public health burdens  We currently understand little about the genetic regulation of muscle health with disease or age  The nematode C  elegans is an established model for understanding the genomic regulation of biological processes of interest  This worm’s body wall muscles display a large degree of homology with the muscles of higher metazoan species  Since C  elegans is a transparent organism, the localization of GFP to mitochondria and sarcomeres allows visualization of these structures in vivo  Similarly, feeding animals cationic dyes, which accumulate based on the existence of a mitochondrial membrane potential, allows the assessment of mitochondrial function in vivo  These methods, as well as assessment of muscle protein homeostasis, are combined with assessment of whole animal muscle function, in the form of movement assays, to allow correlation of sub-cellular defects with functional measures of muscle performance  Thus, C  elegans provides a powerful platform with which to assess the impact of mutations, gene knockdown, and/or chemical compounds upon muscle structure and function  Lastly, as GFP, cationic dyes, and movement assays are assessed non-invasively, prospective studies of muscle structure and function can be conducted across the whole life course and this at present cannot be easily investigated in vivo in any other organism Developmental Biology, Issue 93, Physiology, C  elegans, muscle, mitochondria, sarcomeres, ageing52043Play ButtonImproved Preparation and Preservation of Hippocampal Mouse Slices for a Very Stable and Reproducible Recording of Long-term PotentiationAuthors: Agnès Villers, Laurence Ris  Institutions: University of Mons Long-term potentiation (LTP) is a type of synaptic plasticity characterized by an increase in synaptic strength and believed to be involved in memory encoding  LTP elicited in the CA1 region of acute hippocampal slices has been extensively studied  However the molecular mechanisms underlying the maintenance phase of this phenomenon are still poorly understood  This could be partly due to the various experimental conditions used by different laboratories  Indeed, the maintenance phase of LTP is strongly dependent on external parameters like oxygenation, temperature and humidity  It is also dependent on internal parameters like orientation of the slicing plane and slice viability after dissection 
The optimization of all these parameters enables the induction of a very reproducible and very stable long-term potentiation  This methodology offers the possibility to further explore the molecular mechanisms involved in the stable increase in synaptic strength in hippocampal slices  It also highlights the importance of experimental conditions in in vitro investigation of neurophysiological phenomena Neuroscience, Issue 76, Neurobiology, Anatomy, Physiology, Biomedical Engineering, Surgery, Memory Disorders, Learning, Memory, Neurosciences, Neurophysiology, hippocampus, long-term potentiation, mice, acute slices, synaptic plasticity, in vitro, electrophysiology, animal model50483Play ButtonIn Vivo Modeling of the Morbid Human Genome using Danio rerioAuthors: Adrienne R  Niederriter, Erica E  Davis, Christelle Golzio, Edwin C  Oh, I-Chun Tsai, Nicholas Katsanis  Institutions: Duke University Medical Center, Duke University, Duke University Medical Center Here, we present methods for the development of assays to query potentially clinically significant nonsynonymous changes using in vivo complementation in zebrafish  Zebrafish (Danio rerio) are a useful animal system due to their experimental tractability; embryos are transparent to enable facile viewing, undergo rapid development ex vivo, and can be genetically manipulated 1 These aspects have allowed for significant advances in the analysis of embryogenesis, molecular processes, and morphogenetic signaling  Taken together, the advantages of this vertebrate model make zebrafish highly amenable to modeling the developmental defects in pediatric disease, and in some cases, adult-onset disorders  Because the zebrafish genome is highly conserved with that of humans (~70% orthologous), it is possible to recapitulate human disease states in zebrafish  This is accomplished either through the injection of mutant human mRNA to induce dominant negative or gain of function alleles, or utilization of morpholino (MO) antisense oligonucleotides to suppress genes to mimic loss of function variants  Through complementation of MO-induced phenotypes with capped human mRNA, our approach enables the interpretation of the deleterious effect of mutations on human protein sequence based on the ability of mutant mRNA to rescue a measurable, physiologically relevant phenotype  Modeling of the human disease alleles occurs through microinjection of zebrafish embryos with MO and/or human mRNA at the 1-4 cell stage, and phenotyping up to seven days post fertilization (dpf)  This general strategy can be extended to a wide range of disease phenotypes, as demonstrated in the following protocol  We present our established models for morphogenetic signaling, craniofacial, cardiac, vascular integrity, renal function, and skeletal muscle disorder phenotypes, as well as others  Molecular Biology, Issue 78, Genetics, Biomedical Engineering, Medicine, Developmental Biology, Biochemistry, Anatomy, Physiology, Bioengineering, Genomics, Medical, zebrafish, in vivo, morpholino, human disease modeling, transcription, PCR, mRNA, DNA, Danio rerio, animal model50338Play ButtonDirect Imaging of ER Calcium with Targeted-Esterase Induced Dye Loading (TED)Authors: Samira Samtleben, Juliane Jaepel, Caroline Fecher, Thomas Andreska, Markus Rehberg, Robert Blum  Institutions: University of Wuerzburg, Max Planck Institute of Neurobiology, Martinsried, Ludwig-Maximilians University of Munich Visualization of calcium dynamics is important to understand the role of calcium in cell physiology  To examine calcium dynamics, synthetic fluorescent Ca2+ indictors have become popular  Here we demonstrate TED (= targeted-esterase induced dye loading), a method to improve the release of Ca2+ indicator dyes in the ER lumen of different cell types  To date, TED was used in cell lines, glial cells, and neurons in vitro  TED bases on efficient, recombinant targeting of a high carboxylesterase activity to the ER lumen using vector-constructs that express Carboxylesterases (CES)  The latest TED vectors contain a core element of CES2 fused to a red fluorescent protein, thus enabling simultaneous two-color imaging  The dynamics of free calcium in the ER are imaged in one color, while the corresponding ER structure appears in red  At the beginning of the procedure, cells are transduced with a lentivirus  Subsequently, the infected cells are seeded on coverslips to finally enable live cell imaging  Then, living cells are incubated with the acetoxymethyl ester (AM-ester) form of low-affinity Ca2+ indicators, for instance Fluo5N-AM, Mag-Fluo4-AM, or Mag-Fura2-AM  The esterase activity in the ER cleaves off hydrophobic side chains from the AM form of the Ca2+ indicator and a hydrophilic fluorescent dye/Ca2+ complex is formed and trapped in the ER lumen  After dye loading, the cells are analyzed at an inverted confocal laser scanning microscope  Cells are continuously perfused with Ringer-like solutions and the ER calcium dynamics are directly visualized by time-lapse imaging  Calcium release from the ER is identified by a decrease in fluorescence intensity in regions of interest, whereas the refilling of the ER calcium store produces an increase in fluorescence intensity  Finally, the change in fluorescent intensity over time is determined by calculation of ΔF/F0 Cellular Biology, Issue 75, Neurobiology, Neuroscience, Molecular Biology, Biochemistry, Biomedical Engineering, Bioengineering, Virology, Medicine, Anatomy, Physiology, Surgery, Endoplasmic Reticulum, ER, Calcium Signaling, calcium store, calcium imaging, calcium indicator, metabotropic signaling, Ca2+, neurons, cells, mouse, animal model, cell culture, targeted esterase induced dye loading, imaging50317Play ButtonPreparation of Dissociated Mouse Cortical Neuron CulturesAuthors: Lutz G  W  Hilgenberg, Martin A  Smith  Institutions: University of California, Irvine (UCI) This video will guide you through the process for generating cortical neuronal cultures from late embryo and early postnatal mouse brain  These cultures can be used for a variety of applications including immunocytochemistry, biochemistry, electrophysiology, calcium and sodium imaging, protein and/or RNA isolation  These cultures also provide a platform to study the neuronal development of transgenic animals that carry a late embryonic or postnatal lethal gene mutation  The procedure is relatively straight forward, requires some experience in tissue culture technique and should not take longer than two to three hours if you are properly prepared  Careful separation of the cortical rind from the thalamo-cortical fiber tract will reduce the number of unwanted non-neuronal cells  To increase yields of neuronal cells triturate the pieces of the cortical tissue gently after the enzyme incubation step  This is imperative as it prevents unnecessary injury to cells and premature neuronal cell death  Since these cultures are maintained in the absence of glia feeder cells, they also offer an added advantage of growing cultures enriched in neurons Neuroscience, Issue 10, cellular, molecular, neurobiology, neuron, calcium/sodium imaging, primary cultures, mouse562Play ButtonAnalysis of Schwann-astrocyte Interactions Using In Vitro AssaysAuthors: Fardad T  Afshari, Jessica C  Kwok, James W  Fawcett  Institutions: University of Cambridge Schwann cells are one of the commonly used cells in repair strategies following spinal cord injuries  Schwann cells are capable of supporting axonal regeneration and sprouting by secreting growth factors 1,2 and providing growth promoting adhesion molecules 3 and extracellular matrix molecules 4  In addition they myelinate the demyelinated axons at the site of injury 5 
However following transplantation, Schwann cells do not migrate from the site of implant and do not intermingle with the host astrocytes 6,7  This results in formation of a sharp boundary between the Schwann cells and astrocytes, creating an obstacle for growing axons trying to exit the graft back into the host tissue proximally and distally  Astrocytes in contact with Schwann cells also undergo hypertrophy and up-regulate the inhibitory molecules 8-13 
In vitro assays have been used to model Schwann cell-astrocyte interactions and have been important in understanding the mechanism underlying the cellular behaviour 
These in vitro assays include boundary assay, where a co-culture is made using two different cells with each cell type occupying different territories with only a small gap separating the two cell fronts  As the cells divide and migrate, the two cellular fronts get closer to each other and finally collide  This allows the behaviour of the two cellular populations to be analyzed at the boundary  Another variation of the same technique is to mix the two cellular populations in culture and over time the two cell types segregate with Schwann cells clumped together as islands in between astrocytes together creating multiple Schwann-astrocyte boundaries 
The second assay used in studying the interaction of two cell types is the migration assay where cellular movement can be tracked on the surface of the other cell type monolayer 14,15  This assay is commonly known as inverted coverslip assay  Schwann cells are cultured on small glass fragments and they are inverted face down onto the surface of astrocyte monolayers and migration is assessed from the edge of coverslip 
Both assays have been instrumental in studying the underlying mechanisms involved in the cellular exclusion and boundary formation  Some of the molecules identified using these techniques include N-Cadherins 15, Chondroitin Sulphate proteoglycans(CSPGs) 16,17, FGF/Heparin 18, Eph/Ephrins19 
This article intends to describe boundary assay and migration assay in stepwise fashion and elucidate the possible technical problems that might occur Cellular Biology, Issue 47, Schwann cell, astrocyte, boundary, migration, repulsion2214Play ButtonQuantifying Synapses: an Immunocytochemistry-based Assay to Quantify Synapse NumberAuthors: Dominic M  Ippolito, Cagla Eroglu  Institutions: Duke University, Duke University One of the most important goals in neuroscience is to understand the molecular cues that instruct early stages of synapse formation  As such it has become imperative to develop objective approaches to quantify changes in synaptic connectivity  Starting from sample fixation, this protocol details how to quantify synapse number both in dissociated neuronal culture and in brain sections using immunocytochemistry  Using compartment-specific antibodies, we label presynaptic terminals as well as sites of postsynaptic specialization  We define synapses as points of colocalization between the signals generated by these markers  The number of these colocalizations is quantified using a plug in Puncta Analyzer (written by Bary Wark, available upon request, c eroglu@cellbio duke edu) under the ImageJ analysis software platform  The synapse assay described in this protocol can be applied to any neural tissue or culture preparation for which you have selective pre- and postsynaptic markers  This synapse assay is a valuable tool that can be widely utilized in the study of synaptic development Neuroscience, Issue 45, synapse, immunocytochemistry, brain, neuron, astrocyte2270Play ButtonPreparation of Acute Hippocampal Slices from Rats and Transgenic Mice for the Study of Synaptic Alterations during Aging and Amyloid PathologyAuthors: Diana M  Mathis, Jennifer L  Furman, Christopher M  Norris  Institutions: University of Kentucky College of Public Health, University of Kentucky College of Medicine, University of Kentucky College of Medicine The rodent hippocampal slice preparation is perhaps the most broadly used tool for investigating mammalian synaptic function and plasticity  The hippocampus can be extracted quickly and easily from rats and mice and slices remain viable for hours in oxygenated artificial cerebrospinal fluid  Moreover, basic electrophysisologic techniques are easily applied to the investigation of synaptic function in hippocampal slices and have provided some of the best biomarkers for cognitive impairments  The hippocampal slice is especially popular for the study of synaptic plasticity mechanisms involved in learning and memory  Changes in the induction of long-term potentiation and depression (LTP and LTD) of synaptic efficacy in hippocampal slices (or lack thereof) are frequently used to describe the neurologic phenotype of cognitively-impaired animals and/or to evaluate the mechanism of action of nootropic compounds  This article outlines the procedures we use for preparing hippocampal slices from rats and transgenic mice for the study of synaptic alterations associated with brain aging and Alzheimer's disease (AD)1-3  Use of aged rats and AD model mice can present a unique set of challenges to researchers accustomed to using younger rats and/or mice in their research  Aged rats have thicker skulls and tougher connective tissue than younger rats and mice, which can delay brain extraction and/or dissection and consequently negate or exaggerate real age-differences in synaptic function and plasticity  Aging and amyloid pathology may also exacerbate hippocampal damage sustained during the dissection procedure, again complicating any inferences drawn from physiologic assessment  Here, we discuss the steps taken during the dissection procedure to minimize these problems  Examples of synaptic responses acquired in "healthy" and "unhealthy" slices from rats and mice are provided, as well as representative synaptic plasticity experiments  The possible impact of other methodological factors on synaptic function in these animal models (e g  recording solution components, stimulation parameters) are also discussed  While the focus of this article is on the use of aged rats and transgenic mice, novices to slice physiology should find enough detail here to get started on their own studies, using a variety of rodent models Neuroscience, Issue 49, aging, amyloid, hippocampal slice, synaptic plasticity, Ca2+, CA1, electrophysiology2330Play ButtonMesenteric Artery Contraction and Relaxation Studies Using Automated Wire MyographyAuthors: Lakeesha E  Bridges, Cicely L  Williams, Mildred A  Pointer, Emmanuel M  Awumey  Institutions: North Carolina Central University, Durham, North Carolina Central University, Durham, Wake Forest University School of Medicine Proximal resistance vessels, such as the mesenteric arteries, contribute substantially to the peripheral resistance  These small vessels of between 100-400 μm in diameter function primarily in directing blood flow to various organs according to the overall requirements of the body  The rat mesenteric artery has a diameter greater than 100 μm  The myography technique, first described by Mulvay and Halpern1, was based on the method proposed by Bevan and Osher2  The technique provides information about small vessels under isometric conditions, where substantial shortening of the muscle preparation is prevented  Since force production and sensitivity of vessels to different agonists is dependent on the extent of stretch, according to active tension-length relation, it is essential to conduct contraction studies under isometric conditions to prevent compliance of the mounting wires  Stainless steel wires are preferred to tungsten wires because of oxidation of the latter, which affects recorded responses3 The technique allows for the comparison of agonist-induced contractions of mounted vessels to obtain evidence for normal function of vascular smooth muscle cell receptors 
Medicine, Issue 55, cardiovascular, resistant arteries, contraction, relaxation, myography3119Play ButtonVisualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero ElectroporationAuthors: Emilie Pacary, Matilda A  Haas, Hendrik Wildner, Roberta Azzarelli, Donald M  Bell, Djoher Nora Abrous, François Guillemot  Institutions: MRC National Institute for Medical Research, National Institute for Medical Research, Université de Bordeaux In utero electroporation (IUE) has become a powerful technique to study the development of different regions of the embryonic nervous system 1-5  To date this tool has been widely used to study the regulation of cellular proliferation, differentiation and neuronal migration especially in the developing cerebral cortex 6-8  Here we detail our protocol to electroporate in utero the cerebral cortex and the hippocampus and provide evidence that this approach can be used to study dendrites and spines in these two cerebral regions 
Finally, IUE provides a useful tool to identify functional interactions between genes involved in dendrite, spine and/or synapse development  Indeed, in contrast to other gene transfer methods such as virus, it is straightforward to combine multiple RNAi or transgenes in the same population of cells  In summary, IUE is a powerful method that has already contributed to the characterization of molecular mechanisms underlying brain function and disease and it should also be useful in the study of dendrites and spines Neuroscience, Issue 65, Developmental Biology, Molecular Biology, Neuronal development, In utero electroporation, dendrite, spines, hippocampus, cerebral cortex, gain and loss of function4163Play ButtonImaging Analysis of Neuron to Glia Interaction in Microfluidic Culture Platform (MCP)-based Neuronal Axon and Glia Co-culture SystemAuthors: Haruki Higashimori, Yongjie Yang  Institutions: Tufts University, Tufts Sackler School of Graduate Biomedical Sciences Proper neuron to glia interaction is critical to physiological function of the central nervous system (CNS)  This bidirectional communication is sophisticatedly mediated by specific signaling pathways between neuron and glia1,2   Identification and characterization of these signaling pathways is essential to the understanding of how neuron to glia interaction shapes CNS physiology  Previously, neuron and glia mixed cultures have been widely utilized for testing and characterizing signaling pathways between neuron and glia  What we have learned from these preparations and other in vivo tools, however, has suggested that mutual signaling between neuron and glia often occurred in specific compartments within neurons (i e , axon, dendrite, or soma)3  This makes it important to develop a new culture system that allows separation of neuronal compartments and specifically examines the interaction between glia and neuronal axons/dendrites  In addition, the conventional mixed culture system is not capable of differentiating the soluble factors and direct membrane contact signals between neuron and glia  Furthermore, the large quantity of neurons and glial cells in the conventional co-culture system lacks the resolution necessary to observe the interaction between a single axon and a glial cell 
In this study, we describe a novel axon and glia co-culture system with the use of a microfluidic culture platform (MCP)  In this co-culture system, neurons and glial cells are cultured in two separate chambers that are connected through multiple central channels  In this microfluidic culture platform, only neuronal processes (especially axons) can enter the glial side through the central channels  In combination with powerful fluorescent protein labeling, this system allows direct examination of signaling pathways between axonal/dendritic and glial interactions, such as axon-mediated transcriptional regulation in glia, glia-mediated receptor trafficking in neuronal terminals, and glia-mediated axon growth  The narrow diameter of the chamber also significantly prohibits the flow of the neuron-enriched medium into the glial chamber, facilitating probing of the direct membrane-protein interaction between axons/dendrites and glial surfaces Neuroscience, Issue 68, Molecular Biology, Cellular Biology, Biophysics, Microfluidics, Microfluidic culture platform, Compartmented culture, Neuron to glia signaling, neurons, glia, cell culture4448Play ButtonFluorescence Recovery After Photobleaching (FRAP) of Fluorescence Tagged Proteins in Dendritic Spines of Cultured Hippocampal NeuronsAuthors: Chan-Ying Zheng, Ronald S  Petralia, Ya-Xian Wang, Bechara Kachar  Institutions: National Institutes of Health, Bethesda FRAP has been used to quantify the mobility of GFP-tagged proteins  Using a strong excitation laser, the fluorescence of a GFP-tagged protein is bleached in the region of interest  The fluorescence of the region recovers when the unbleached GFP-tagged protein from outside of the region diffuses into the region of interest  The mobility of the protein is then analyzed by measuring the fluorescence recovery rate  This technique could be used to characterize protein mobility and turnover rate 
This FRAP protocol shows how to perform a basic FRAP experiment as well as how to analyze the data Neuroscience, Issue 50, Spine, FRAP, hippocampal neurons, live cell imaging, protein mobility2568Play ButtonPrimary Neuronal Cultures from the Brains of Late Stage Drosophila PupaeAuthors: Beatriz Sicaeros, Jorge M  Campusano, Diane K  O'Dowd  Institutions: University of California, Irvine (UCI) In this video, we demonstrate the preparation of primary neuronal cultures from the brains of late stage Drosophila pupae  The procedure begins with the removal of brains from animals at 70-78 hrs after puparium formation  The isolated brains are shown after brief incubation in papain followed by several washes in serum-free growth medium  The process of mechanical dissociation of each brain in a 5 ul drop of media on a coverslip is illustrated  The axons and dendrites of the post-mitotic neurons are sheered off near the soma during dissociation but the neurons begin to regenerate processes within a few hours of plating  Images show live cultures at 2 days  Neurons continue to elaborate processes during the first week in culture  Specific neuronal populations can be identified in culture using GAL4 lines to drive tissue specific expression of fluorescent markers such as GFP or RFP  Whole cell recordings have demonstrated the cultured neurons form functional, spontaneously active cholinergic and GABAergic synapses  A short video segment illustrates calcium dynamics in the cultured neurons using Fura-2 as a calcium indicator dye to monitor spontaneous calcium transients and nicotine evoked calcium responses in a dish of cultured neurons  These pupal brain cultures are a useful model system in which genetic and pharmacological tools can be used to identify intrinsic and extrinsic factors that influence formation and function of central synapses 
Context 3: HOFFMAN: I'm delighted to introduce the chair of the last session, Mara Liasson from the National Public Radio  Mara is Congressional correspondent for NPR, and covers activities in Congress in D C  Right now, this week, she has been covering the tax bill, which people currently are going at hot and heavy  She took time off from her busy schedule to come here to help us sort out some of these key issues for today, and more importantly, for what happens in the next decade and beyond  I'll turn it over to Mara to get the panel going 
LIASSON: Thank you very much  I am probably the only person here who has absolutely no background in technology  Anyway, I am the only one who does not understand what the panelists are going to be talking about (laughter), and although they have already told me that they do not appreciate people who think that that's a great quality and look down on people who are technical, and I certainly do not, I will reserve the right to insist that they all talk in terms that people like me can understand, since there is more of me out there than you, although not in this room today  (laughter) What we are going to do is introduce each panelist, and each one will make a short three- to five-minute presentation  Then my instructions say that we are going to have a McLaughlin Group discussion, which I guess means lots of yelling and screaming and talking at once  (laughter) After that's over, about 4:10, we'll open up the panel for questions from the audience 
To my left is Peter Denning, who is Chairman of the Computer Science Department at George Mason University and also the associate dean for computing  He is the program chair of this conference, has also served as the president of ACM, and he is currently the editor of Communications 
Simon Davies, to my right, also wears blue suits, but you can tell him from Mitch, because he wears a white hat  (laughter) He is from Sydney, Australia, and is the Director General of Privacy International, which is an international network of privacy advocates  He is also an author, a journalist, and radio commentator 
To his right is Roland Homet  He is an information policy writer and thinker who recently opened his own public policy writing firm here in Washington -- it's called Executive Ink, not Inc , as it is written in your programs, so you can scratch that out 
Esther Dyson, at the end of the panel, is among the most respected commentators on developing technology trends in the personal computer business  She publishes two newsletters, Release 1 0 and Rel-EAST  She has also been one of the driving forces promoting East-West relations through computer networks  She is a board member of the Electronic Frontier Foundation as well 
I'll ask Peter to start 
P  DENNING: Thank you  Starting around 1850, people of many countries looked to their governments to regulate commerce, erase inequities, and build societies of better human beings  For over a hundred years, many people, from peasants to intellectuals, had faith that strong governments would bring them a better life  This faith was part of the clearing in which Communist governments flourished; although the United States took an anti-Communist stand, the same faith fostered a strong government that promised salvation by great national programs including Social Security, welfare, food stamps, the War on Poverty, and the Great Society  This faith is now shattered  People no longer trust that powerful government can deliver a better life 
The dramatic collapse of Communism in Eastern Europe and the Soviet Union illustrates this, as does the growing disillusionment of the American people for federal, state, and local governments  The poor track record of government is not the only reason for the shift  Information technology has accelerated the process  Communications that took weeks in the last century now take fractions of a second  Business success depends on what happens around the globe, not only on local conditions  Radio, TV, fax, and now E-mail are common worldwide, so much so that not even a powerful government can control what information its citizens have  Because the space of opportunity for people to engage in transactions with each other has been so enormously enlarged during the past decade, faith in marketplace democracies is on the rise worldwide; correspondingly faith in central management mechanisms is on the decline  This shift has brought with it a shift of the power of institutions  Government institutions tend to try to hold onto their power by regulatory coercion to enforce the old ways  This can produce big tensions and even promote breakage 
Nowhere can this be seen more clearly than in the cryptographic area which we have just been talking about in the previous hour  This technology, cryptography, produces mechanisms for digital signatures, authentication, electronic money, certificates, and private communication -- all offering a way for standard business practices now based on paper to be shifted into the electronic media  The success of worldwide enterprises depends on this shift being completed rapidly and effectively  As more people realize this, the momentum for incorporating cryptographic technology into the information infrastructure is accelerating 
In this country, the National Security Agency has long been given the authority to regulate cryptography  This authority was granted in another time when the success of the country depended upon the ability of its government to gather intelligence and communicate in secret  These premises made sense in a world where most of the power resided in governments, but the world is changing  Much economic power is now accumulating in large apolitical transnational corporations  These corporations place their own concerns and strategies ahead of those of governments of the countries in which they do business  Like governments, they are interested in gathering intelligence about competitors and in conducting business in private  Unlike governments, they want open access to the technologies of authentication, electronic money, digital signatures, and certificates that will allow them to conduct business transactions across the network  So it is no longer true that national power and national security are increased when government has the sole right to gather intelligence and encipher communications  Now the strength of a country depends not only on its government, but also on its corporations  The old premises have fallen away in the new reality, but the old policy remains  It's time to rethink the policy, before tensions between a threatened government and corporations produce significant social tension and perhaps breakage 
Well, digital media -- computer-based communications -- are the printing press of the 21st century, and as the printing press transformed society, created the modern individual, gave rise to the basis of the democratic state and to the notion of individual rights, I suspect that we will see a similar, radical transformation of the very constitution of global society in the next century, facilitated by this enabling technology  I would be the last person to try to sketch out the details, or tell you what the issues are going to be, but I want to share with you some feelings about what is really going to matter, as we go about this -- and I'll start with something about myself 
You see a guy wearing a suit; most of you know I have a lot of money -- I'm a successful businessman  God knows what images propagate around the media and settle in people's minds, but I've always seen myself, and felt myself to the core of my being, as an outsider, every bit as much as a self-proclaimed outsider, as Tom Jennings -- who spoke so eloquently about this at the Pioneer awards* yesterday -- was  *The Electronic Freedom Foundation presented its first awards at a related, adjacent reception which was not formally a part of the conference 
I think we are all outsiders; we are all different, all unique  We're not the same  We share an underlying common humanity, but we should not be asked to subjugate ourselves to some form of mass society that causes us each to become indistinguishable from one another  I believe that computer- based communications technology is an enabling technology to liberate individuals and to free us from the oppressive influence of large institutions, whether those are public or private  And I am talking about an economic restructuring that results in a much more decentralized society, and social restructuring in an affirmation of the simple right to be left alone  I think Cyberspace is good for individuals, and I think that's important  I also think that the flip side of the coin, the creation of community, which we so sorely lack in this country today, can be facilitated through these technologies 
I have experienced that for myself, as many of you have on your various computer networks on conferencing systems like the WELL  It is enormously liberating to overcome the artificial boundaries of space and time  We are prisoners of geography in the physical world, and our communities are largely a product of who we can see face to face each day, even though our real comrades and colleagues may be scattered all over the world and our interests -- whether they are hobbies or political interests or religious interests, whatever they might be -- can be facilitated if we are able to get in touch with, to form bonds with, to exchange views and ideas with other kindred spirits  And I believe this technology is an enabling technology for the formation of community  My hope is that we will have the wisdom to create policies which enable individuals to flourish free from the chains of mass society, and which enable voluntary communities of people, individuals, groups who come together to be with each other and to work together  I hope both of those become possible 
DAVIES: I feel very warmed by the various visions of the future that have come out of this conference, but I am a cynic, and cynicism is good, because it adds fiber  (laughter) How nice the world would be if everyone was like Mitch, but they're not, because the future is in the hands of ruthless, greedy little men 
I want to paint the vision of the future that I have, and I hope it's not too depressing because there is a future, a good future    possibly  I agree, as many of you do, that the future is going to be like some giant informational Yggdrasil* *Reference from Old Norse mythology -- the Yggdrasil was a giant ash tree whose roots held together the universe   We'll all be part of interconnectivity, the likes of which we can scarcely imagine right now  I imagine it will be like an organism where we're independent and interdependent, and so it's like a two-edged sword  That's all very nice, and we can see that we form part of that new community  But, I see a world with 15 billion beings scrambling for life, where four-fifths of the world lives on half a liter of water a day, where people grow up to see their children dying, where new political frontiers are destroying freedoms and the democracy that we have developed over the last two centuries  I see a world where there is very little hope for nearly everybody on the planet, except for the elite -- that's us -- except for those of us who are plugged into the informational Yggdrasil 
What I see is that 14 of those 15 billion people are a lot of pissed-off people who have their eyes set on what they see, not as a wonderful informational community, but as the beast  And they see that that is where the resources are, and that's where the opportunities are, and that's where the political power is  I can't see a future for us in a world where ultimately the great demon becomes information  It might be good for us, but for the disaffected four-fifths of the world, information is going to be something which, frankly, we can do without, because in a world with almost no resources left, surely information is selfishness 
HOMET: Thank you  I'm grateful to the organizers for including me in these proceedings -- they are reminiscent for me of some information policy conferences that I organized 15 to 20 years ago for the Aspen Institute  The particulars have certainly changed, but the dynamics remain much the same  For me, these are well-represented by Peter Denning's image of a changeable clearing in the woods  At any given time, as I see it, the clearing is an acceptable standoff between the forces of modernization and of traditional culture, between freedom and discipline, between structure and spontaneity  Now we voice these as opposites, but in fact, they need each other  It is the creative tension between technological innovation and established order that allows society to hold together and progress to take place  Take away freedom and order will be overthrown -- witness the Soviet Union  Take away tradition, and modernization will be crushed -- witness Iran  The clearing must be respected and it must move  Just as Benjamin Cardozo of the U S  Supreme Court said 65 years ago, the genius of the American system is its penchant for ordered liberty  When both halves of the equation work against each other and together in Hegelian terms, the clearing that they produce is, at any given time, a prevailing hypothesis, which is challenged by a new antithesis  Together they can produce a fresh synthesis  And all that is very familiar  What is new and trying is the sweep and pace of innovation today, plus -- and this is what we sometimes forget -- the political volatility of the value systems that this can induce  If you doubt that, consider the Buchanan campaign and what's been going on with the Endowment for the Arts and public broadcasting  These are signs of people running scared, and they can cause damage 
So the answer for the 21st century is to proceed under power, but with restraint, to practice what Mitch Kapor in another connection called toleration for opposing forces and perspectives  We need each other to keep the enterprise together and on course  For computer practitioners represented in this room, this means restraint from provoking unnecessary and damaging social backlash  A good example might be New York telcos offering free per-call and per-line blocking with this caller identification service  For regulators and law enforcers, restraint means asking, "Do you know enough to freeze emerging conduct in a particular form or pattern?" I was very taken by the role reversal exercise organized by Michael Gibbons on Wednesday night  It led me to wonder what might have happened to the government's wiretapping and encryption proposals had they been subjected to a comparable advanced exercise before introduction 
Sixteen years ago in Aspen, Colorado, I convened a gathering of federal policymakers and invited them to consider a suggested matrix of policy values and processes in the information society  The first two of those values -- it will not surprise you to know -- were freedom of discourse and individual privacy  But there were more: freedom of economic choice is one; the general welfare another; popular sovereignty, worth pausing on, I described as avoiding concentrations of economic and political power in any sector of industry or government that impinge unduly on the freedoms or welfare of the citizenry  And then there is progress, social progress, the fostering, I said, of market incentives and opportunities for technological and service innovations and for widened consumer choice among technologies and services  Now obviously if you give just a moment's thought to it, you will recognize, as I think we have in this conference, that these values can collide with each other at key points, and therefore accommodations must be made  For that we need processes of accommodation  I also suggested some of those  After you identify the relevant values and goals, you then should ask yourself about the necessity and the appropriateness of having government make any decision on the matter  And this has to do with such things like the adequacy of decision-making standards, the availability of adequate information, and the adequacy of personnel resources to deal with it  Then you get into dividing up the possible roles of the various elements of government -- the regulatory agencies, the Executive Branch, the Judiciary, and the Congress  It doesn't stop there, because you need to ask about international implications, which we have done some of here  And federal/state implications -- very often allowing the state to make a stab at social ordering in the first instance is, as Justice Brandeis often said, the best way, through the social laboratory technique, to try out what is the right answer, without endangering the whole society  And as we have heard today, we need also to think about the availability of non-coercive instruments of accommodation, like a federal data protection board 
DYSON: I want to just say one thing about this business of crypto technology -- it is a very simple sentence, and everyone seems to slip slightly by it; that is, if you outlaw guns, only outlaws will have guns  Crypto technology is fundamentally a defensive weapon  It may protect murderers and thieves, but it is not a weapon that murders, kills, does anything bad; and so it is a very different kettle of fish from any other kind of weapon  The whole point is that information is powerful, and that the free flow of information, privacy-protected, empowers the powerless and is dangerous to the powerful -- and that's why we need our privacy protected 
Now let me just talk a wee bit about the future  A couple of days ago, a reporter called me and asked what the EFF stood for  I kind of floundered around and said, "Well, we want privacy, we want good hackers to be protected and bad crackers to be punished  We want people to understand the difference, and we want all these good things, but we really don't want to grab power " The guy kept on not quite getting it  The real answers were pro choice



ground_truth: They are rich in actin and have been shown to be highly dynamic.

prompt_front: Now, answer the following question based on the text below, only give me the answer and do not output any other words.

Question: What are some reasons for the lack of data sharing in archaeobotany?

Read the following text and answer briefly.

Context 2: Sowing the Seeds of Future Research: Data Sharing, Citation and Reuse in Archaeobotany
Reading: Sowing the Seeds of Future Research: Data Sharing, Citation and Reuse in Archaeobotany
University of Oxford, GB
Lisa is a post-doctoral research fellow at All Souls College, University of Oxford  Her publications include the co-authored volume The Rural Economy of Roman Britain (Britannia Monographs, 2017)  Her research interests are focussed on agricultural practices in the later prehistoric and Roman period and the utilisation of archaeobotanical data to investigate human-plant relationships 
The practices of data sharing, data citation and data reuse are all crucial aspects of the reproducibility of archaeological research  This article builds on the small number of studies reviewing data sharing and citation practices in archaeology, focussing on the data-rich sub-discipline of archaeobotany  Archaeobotany is a sub-discipline built on the time-intensive collection of data on archaeological plant remains, in order to investigate crop choice, crop husbandry, diet, vegetation and a wide range of other past human-plant relationships  Within archaeobotany, the level and form of data sharing is currently unknown  This article first reviews the form of data shared and the method of data sharing in 239 articles across 16 journals which present primary plant macrofossil studies  Second, it assesses data-citation in meta-analysis studies in 107 articles across 20 journals  Third, it assesses data reuse practices in archaeobotany, before exploring how these research practices can be improved to benefit the rigour and reuse of archaeobotanical research 
Keywords: Archaeobotany, Data reuse, Data sharing, Open science
How to Cite: Lodwick, L , 2019  Sowing the Seeds of Future Research: Data Sharing, Citation and Reuse in Archaeobotany  Open Quaternary, 5(1), p 7  DOI: http://doi org/10 5334/oq 62
Accepted on 29 May 2019 Submitted on 25 Mar 2019
Archaeology is a discipline built on the production and analysis of quantitative data pertaining to past human behaviour  As each archaeological deposit is a unique occurrence, ensuring that the data resulting from excavation and analysis are preserved and accessible is crucially important  Currently, there is a general perception of a low level of data sharing and reuse  Such a low level of data availability would prevent the assessment of research findings and the reuse of data in meta-analysis (Kansa & Kansa 2013; Moore & Richards 2015)  As observed across scientific disciplines, there is a major problem in the reproduction of scientific findings, commonly known as the ‘replication crisis’ (Costello et al  2013)  A range of intersecting debates contribute to this, including access to academic findings (open access), open data, access to software and access to methodologies, which can be broadly grouped as open science practices  Without these, the way that scientific findings can be verified and built upon is impaired  Questions of reproducibility have been raised in recent years in archaeology, with considerations of a range of practices which can improve the reproducibility of findings, and a recent call for the application of open science principles to archaeology (Marwick et al  2017)  Discussion has so far focussed on access to grey literature (Evans 2015), data sharing (Atici et al  2013), data citation practices (Marwick & Pilaar Birch 2018) and computational reproducibility (Marwick 2017), with a focus on lithics, zooarchaeological evidence, and archaeological site reports 
Quantitative assessments of current levels of data sharing, data citation and reuse remain limited in archaeology  The focus of evaluation has been on the uptake of large-scale digital archives for the preservation and dissemination of digital data, such as the Archaeology Data Service (ADS), utilised by developer-led and research projects, and recommended for use by many research funders in the UK (Richards 2002; Wright and Richards 2018)  Much less focus has been paid to the data-sharing practices of individuals or small-groups of university-based researchers who may be disseminating their research largely through journal articles  Recent work on the availability of data on lithics assemblages found a low level of data sharing (Marwick & Pilaar Birch 2018) and there are perceptions of low levels of data reuse (Huggett 2018; Kintigh et al  2018)  Within zooarchaeology numerous studies have explored issues of data sharing and reuse (Kansa & Kansa 2013, 2014), and the sub-discipline is seen as one of the most advanced areas of archaeology in regards to open science (Cooper & Green 2016: 273)  Beyond zooarchaeology, however, explicit discussion has remained limited 
This paper assesses data sharing and reuse practices in archaeology through the case study of archaeobotany – a long established sub-discipline within archaeology which has well-established principles of data recording  Archaeobotany is an interesting case study for data sharing in archaeology as it straddles the division of archaeology between scientific and more traditional techniques  Quantitative data on archaeological plant remains are also of interest to a range of other fields, including ecology, environmental studies, biology and earth sciences  The key issues of data sharing and data reuse (Atici et al  2013) have been touched upon in archaeobotany over the past decade within broader discussions on data quality (Van der Veen, Livarda & Hill 2007; Van der Veen, Hill & Livarda 2013)  These earlier studies focussed on the quality and availability of archaeobotanical data from developer-funded excavations in Britain and Cultural Resource Management in North America (Vanderwarker et al  2016: 156)  However, no discussion of data-sharing and reuse in academic archaeobotany occurred  A recent review of digital methods in archaeobotany is the notable exception, with discussions of the challenges and methods of data sharing (Warinner & d’Alpoim Guedes 2014) 
Currently, we have no evidence for the levels of data sharing and reuse within archaeobotany  This article provides the first quantitative assessment of 1) data publication in recent archaeobotanical journal articles 2) data citation in recent archaeobotanical meta-analysis 3) the reuse of archaeobotanical datasets, in order to assess whether practices need to change and how such changes can take place 
2  Data Publication and Re-use Practices in Archaeobotany
2 1  History of data production and publication
Archaeobotanical data falls within the category of observational data in archaeology (Marwick & Pilaar Birch 2018)  Archaeobotanical data is considered as the quantitative assessment of plant macrofossils present within a sample from a discrete archaeological context, which can include species identification, plant part, levels of identification (cf  – confer or “compares to”), and a range of quantification methods including count, minimum number of individuals, levels of abundance and weight (Popper 1988)  Archaeobotanical data is usually entered into a two-way data table organised by sample number  Alongside the counts of individual taxa, other information is also necessary to interpret archaeobotanical data, including sample volume, flot volume, charcoal volume, flot weight, level of preservation, sample number, context number, feature number, feature type and period  Beyond taxonomic identifications, a range of other types of data are increasingly gathered on individual plant macrofossils (morphometric measurements, isotopic values, aDNA) 
Archaeobotanical training places a strong emphasis on recording data on a sample-by-sample basis (Jacomet & Kreuz 1999: 138–139; Jones & Charles 2009; Pearsall 2016: 97–107)  Time-consuming methodologies utilised in the pursuit of accurate sample-level data recording include sub-sampling and splitting samples into size fractions and counting a statistically useful number of items per sample (Van der Veen & Fieller 1982)  The creation of sample-level data means analysis is often undertaken on the basis of individual samples, for instance the assessment of crop-processing stages and weed ecological evidence for crop husbandry practices  The analysis of sample level data also enables archaeobotanical finds to be integrated alongside contextual evidence from archaeological sites  Requirements for the publication of this data are in place in some archaeological guidelines, for instance current Historic England guidelines for archaeological practice in England (Campbell, Moffett & Straker 2011: 8) 
From the earliest archaeobotanical reports, such as Reid’s work at Roman Silchester, the sample from which plant remains were recovered was noted (Lodwick 2017a), but often results were reported as a list of taxa, or long catalogues of detailed botanical descriptions with seed counts, such as Knörzer’s work at Neuss (Knörzer 1970)  Early systematic archaeobotanical reports displayed data within in-text tables, for example Jones’s work at Ashville (Jones 1978) and the two-way data table has been the standard form of reporting archaeobotanical data ever since  Often data tables are presented within book chapters or appendices, but the financial, space and time constraints of book publishing are limiting  Furthermore, there is the perception that specialist data was not necessary for publication (Barker 2001)  Hence, alternative methods of the dissemination of specialist archaeological data were pursued in the later twentieth century 
From the 1980s, archaeobotanical data tables were often consigned to microfiche following a Council for British Archaeology and Department of Environment report (Moore & Richards 2015: 31), with the example of the excavation of Roman Colchester where the contents of all archaeobotanical samples were available on microfiche (Murphy 1992)  An alternative in the 2000s was providing data tables on CD Rom as seen, for instance, in the CD accompanying the study of a Roman farmstead in the Upper Thames Valley (Robinson 2007) or the One Poultry excavations in London (Hill and Rowsome 2011)  Meanwhile, the inception of the Archaeology Data Service, a digital repository for heritage data, in 1996 meant archaeological datasets were increasingly digitally archived, for instance the data from the Channel Tunnel Rail Link Project (Foreman 2018) or a recent large-scale research excavation at Silchester (University of Reading 2018)  In these cases, archaeobotanical data is available to download as a  csv file 
Whilst the data publication strategy of large excavations was shifting, the availability of data from post-excavation assessment reports has remained challenging  So-called ‘grey literature’ results from the initial evaluation stage of developer-funded investigations and accompanying post-excavation assessment often contain a semi-quantitative evaluation of archaeobotanical samples on a scale of abundance  Whilst paper reports were initially deposited with county Historic Environment Records, a process of digitisation focussing on the Roman period has meant many pdfs are now available through the ADS (Allen et al  2018), whilst born-digital reports are now deposited through OASIS (Online AccesS to the Index of archaeological investigationS), as part of the reporting process (Evans 2015), althought the extent to which specialist appendices are included is variable 
These varying ‘publication’ strategies means archaeobotanical data is often available somewhere for recent developer-funded excavations and large-scale developer-funded excavations, even if much of this data is as a printed table or  pdf file (Evans 2015; Evans and Moore 2014)  However, academic journals are typically perceived as the most high-status publication venue for archaeobotanical data, and a crucial publication venue for academics in order to comply with institutional requirements and the norms of career progression  Aside from the problem of access to pay-walled journals by those without institutional subscriptions to all journals, the publication of primary data alongside research articles faces various problems, from the outright lack of inclusion of data, to problematic curation of supplementary data and a lack of peer review of data (Costello et al  2013; Warinner and d’Alpoim Guedes 2014: 155; Whitlock, 2011)  The extent of these problems for archaeobotany is currently unknown  Given the growth in archaeobotanical data production as methodologies are introduced into many new regions and periods over the last decade, it is vital that we know whether the mass of new data being produced is made available and is being reused 
Recent important advances within archaeobotanical data sharing have focussed on the construction of the ARBODAT database, developed by Angela Kreuz at the Kommission für Archäologische Landesforschung in Hessen  The database is used by a range of researchers in Germany, the Czech Republic, France and England (Kreuz & Schäfer 2002)  Data sharing enabled by the use of this database has facilitated research on Neolithic agriculture in Austria, Bulgaria and Germany (Kreuz et al  2005), and Bronze Age agriculture in Europe (Stika and Heiss 2012)  The use of this database makes data integration between specialists easier due to the shared data structure and metadata description, but often the primary archaeobotanical data is not made publicly available 
2 2  Meta-analysis in archaeobotany
Beyond the need to preserve information, a key reason for the formal sharing of archaeobotanical data is in its reuse to facilitate subsequent research  There has been a long-standing concern within archaeobotany with the need to aggregate datasets and identify temporal and spatial patterns  The palaeobotanist Clement Reid maintained his own database of Quaternary plant records in the late nineteenth century (Reid 1899), which formed the foundation of Godwin’s Quaternary database (Godwin 1975)  Mid-twentieth century studies of prehistoric plant use compiled lists of archaeobotanical materials incorporating full references and the location of the archive (Jessen & Helbaek 1944)  The International Work Group for Palaeoethnobotany was itself founded in 1968 in part with the aim to compile archaeobotanical data, first realised through the publication of Progress in Old World Palaeoethnobotany (Van Zeist, Wasylikowa & Behre 1991), and subsequently through the publication of annual lists of new records of cultivated plants (Kroll 1997) 
To take England as an example, regional reviews produced by state heritage authorities have provided catalogues of archaeobotanical datasets in particular time periods and regions (e g  Murphy 1998)  When one archaeobotanist has undertaken the majority of study within a region, pieces of synthesis within books have provided a relatively comprehensive review, for instance in the Thames Valley, UK (Lambrick & Robinson 2009)  Over the last decade regional synthesis has occurred within several funded reviews which produced catalogues of sites with archaeobotanical data (Lodwick 2014; McKerracher 2018; Parks 2012) and a series of funded projects in France have enabled regional synthesis (Lepetz & Zech-Matterne 2017)  However, many of these reviews are not accompanied by an available underlying database, and draw upon reports which are themselves hard to access 
Through the 1990s and 2000s, a series of databases were constructed in order to collate data from sites in a particular region and facilitate synthetic research  However, these databases have all placed the role of data archiving onto later projects specifically funded to collate data, rather than sourcing datasets at the time of publication  Such a model is unsustainable, and is unlikely to result in all available datasets being compiled  The Archaeobotanical Computer Database (ABCD), published in 1996 in the first issue of Internet Archaeology, contained much of the archaeobotanical data from Britain available at the time of publication, largely at the level of individual samples  The database was compiled between 1989 and 1994 and is still accessible through the accompanying online journal publication (Tomlinson & Hall 1996)  The ABCD made major contributions to recent reviews of the Roman and Medieval periods (Van der Veen, Livarda & Hill 2008; Van der Veen, Hill & Livarda 2013)  However, the database could only be centrally updated, with the online resource remaining a static version, lacking much of the new data produced subsequent to the implementation of PPG16 in 1990  The ADEMNES database, created through a research project undertaken at the Universities of Freiburg and Tübingen, contains data from 533 eastern Mediterranean and Near Eastern sites (Riehl & Kümmel 2005)  Kroll has maintained the Archaeobotanical Literature Database to accompany the Vegetation History and Archaeobotany articles (Kroll 2005) now accessible as a database (Kirleis & Schmültz 2018)  Numerous other databases have collated archaeobotanical studies, including the COMPAG project (Fuller et al  2015), the Cultural Evolution of Neolithic Europe project (Colledge 2016), RADAR in the Netherlands (van Haaster and Brinkkemper 1995), BRAIN Botanical Records of Archaeobotany Italian Network (Mercuri et al  2015) and CZAD – Archaeobotanical database of Czech Republic (CZAD 2019) 
The majority of databases have a restricted regional coverage, whilst research-project driven period-specific databases provide overlapping content  Whilst there are a wide range of archaeobotanical databases available, few contain primary datasets (other than the ABCD) which can be downloaded as  csv files  Data which is most commonly available are bibliographic references per site, with some indications of mode of preservation, quantity of archaeobotanical data, and sometimes taxa present  The databases do not inter-relate to each other, and function primarily as bibliographic sources enabling researchers to find comparative sites or to identify published datasets which need to be re-tabulated prior to meta-analysis  The IWGP website curates a list of resources, but otherwise the resources are often disseminated through the archaeobotany jiscmail list 
Beyond the aim of cataloguing archaeobotanical data within a region and period, meta-analysis is often used in archaeobotany to identify spatial and chronological trends in a range of past human activities, for instance crop choice, crop husbandry practices, plant food consumption, the trade in luxury foods or the use of plants in ritual  Meta-analysis can be undertaken on the basis of simple presence/absence data per site, but in order for such analysis to be rigorous and comparable, sample-level data must be utilised  For instance, sample-level data is required for meta-studies, in order to identify high-quality samples of unmixed crops for weed ecology analysis (Bogaard 2004), to assess the importance of context in the evaluation of wild plant foods (Wallace et al  2019), or to use volumetric measurements as a proxy for scale (Lodwick 2017b)  The reuse of archaeobotanical data also extends to include datasets used as “controls” in commonly used forms of statistical analysis, for instance Jones’s weed data from Amorgos, Greece, which is utilised as a control group in discriminant analysis of crop-processing stage (Jones 1984), and ethnographic observations of crop items in different crop-processing stages (Jones 1990) 
2 3  Open data principles and solutions
Debates over issues of data publication and meta-analysis have been on-going across scientific disciplines over the last decade (Editors 2009), and have been summarised within principles of open science, as recently set out in relation to archaeology (Marwick et al  2017)  Open Data is one of the three core principles for promoting transparency in social science (Miguel et al  2014)  The FAIR principles, developed by representatives from academia, industry, funding agencies, industry and publishers, provide four principles which data sharing should meet for use by both humans and machines – Findability, Accessibility, Interoperability, and Reusability (Wilkinson et al  2016)  A recent report assessing the adoption and impact of FAIR principles across academia in the UK included archaeology as a case study (Allen and Hartland 2018: 46)  It reported how the ADS was often used to archive data, but that “The journal itself provides the “story” about the data, the layer that describes what the data is, how it was collected and what the author thinks it means ” The report also raises the problem that smaller projects may not have the funding to utilise the ADS, meaning that other repositories are utilised  Increasingly, archaeological data is made available through a wide range of data repositories (OSF, Mendeley Data, Zenodo, Open Context), university data repositories (e g  ORA-Data), or social networking sites for academics (Academia edu, ResearchGate)  More widely in archaeology, some have observed that archaeological data is rarely published (Kintigh et al  2014), and recent reviews have reported low levels of data sharing (Huggett 2018; Marwick & Pilaar Birch 2018)  A closely related issue is that of data reuse  Responsible reuse of primary data encourages the sharing of primary data (Atici et al  2013), but levels of data reuse in archaeology are thought to remain low (Huggett 2018)  Principles for responsible data citation in archaeology have recently been developed summarising how datasets should be cited (Marwick & Pilaar Birch 2018) 
In order to assess the current status of data sharing, citation and data re-use in archaeobotany, a review was undertaken of the publication of primary data and the publication of meta-analysis in major archaeological journals over the last ten years, building on recent pilot studies within archaeology (Marwick & Pilaar Birch 2018)  The review of academic journals provided a contrast to recent assessments of archaeobotanical data deriving from developer-funded archaeology (Lodwick 2017c; Van der Veen, Hill & Livarda 2013)  Journal articles have been selected as the focus of this study as the provision of online supplementary materials in the majority of journals and the ability to insert hyperlinks to persistent identifiers (eg a DOI) to link to datasets available elsewhere should not limit the publication of data and references  Much archaeobotanical data is also published elsewhere, especially from projects not based in the university sector, that is commercial or community archaeology in the UK  Archaeobotanical datasets emanating from this research are more commonly published through monographs, county journal articles, and unpublished (or grey literature) reports, but these are beyond the scope of the current review 
All journal articles were included which represent the principle reporting of a new archaeobotanical assemblage  The selected journals fall within three groups  First, what is considered the specialist archaeobotanical journal (Vegetation History and Archaeobotany (VHA))  Second, archaeological science journals (Archaeological and Anthropological Sciences, Environmental Archaeology, The Holocene, Journal of Archaeological Science (JAS), Journal of Archaeological Science: Reports (JASR), Journal of Ethnobiology, Quaternary International, Journal of Wetland Archaeology), which can be considered as specialist sub-disciplinary journals which should be maintaining data-quality  Third, general archaeology journals (Antiquity, Journal of Field Archaeology, Oxford Journal of Archaeology, Journal of Anthropological Archaeology, Journal of World Prehistory)  Finally, the broader cross-disciplinary journals PLoS One and Proceedings of the National Academy of Sciences (PNAS) were included  Published articles from the past ten years (2009–2018) have been analysed in order to assess the availability of plant macrofossil data  This ten-year period brackets the period where most archaeological journals have moved online and adopted supplementary materials 
Data citation in synthetic studies has been assessed in the same range of publications  The extent of data reuse ranges from the analysis of whole sample data to the presence/absence of individual crops  The location of a data citation has been assessed in the same range of publications, with the addition of journals where occasional research incorporating archaeobotanical data is featured (Britannia, Journal of Archaeological Research, Ethnobiology Letters, Medieval Archaeology, Proceedings of the Prehistoric Society, World Archaeology)  The underlying dataset for the analysis is available in Lodwick 2019 
4 1  Primary data sharing
Here, the location of primary archaeobotanical data, that is sample level counts of macroscopic plant remains, was assessed for 239 journal articles across 16 journals (Lodwick 2019 Table 1)  Figure 1 shows the results grouped by journal  Overall, only 56% of articles shared their primary data  In, Antiquity, JAS, JASR, PLOS One, Quaternary International and VHA, the highest proportion of publications did not include their primary data, that is to say that the sample-by-sample counts of plant macrofossils was not available  This level of data is comparable to the findings of other pilot studies in archaeology  Marwick and Pilaar Birch found a data sharing rate of 53% from 48 articles published in Journal of Archaeological Science in Feb – May 2017 (Marwick & Pilaar Birch 2018: 7), and confirm previous assertions that data is often withheld in archaeology (Kansa 2012: 499)  This is better than some disciplines, with a 9% data sharing rate on publication found across high impact journal science publications (n = 500) (Alsheikh-Ali et al  2011) and 13% in biology, chemistry, mathematics and physics (n = 4370) (Womack 2015), yet still indicates that nearly half of articles did not include primary data  Primary archaeobotanical data is more likely to be shared in archaeobotanical and archaeological science journals than general archaeology journals  However, within the primary archaeobotanical journal, VHA, 51% of articles do not include their primary data (Figure 1) 
Chart showing the location of primary archaeobotanical data by journal in primary archaeobotanical data publications 
Where primary data was not shared, the data which was available ranged from summary statistics, typically counts or frequencies, reported either by site, site phase, or feature group  Figure 2 summarises these results by year, showing that there is a gradient within articles not sharing their full ‘raw’ data, from those only provided sample counts on one aspect of the archaeobotanical assemblage, to those only presenting data graphically or within discussion  Beyond full data, the most common form of data shared is either summary counts per site or summary counts per feature or phase  Whilst this data does enable some level of reuse, the results of any sample-level data analysis presented within an article cannot be verified, and the data cannot be reused for crop-processing or weed ecology analysis which requires sample level data  Furthermore, such data would have been collected on a sample-by-sample basis, but this information is lost from the resulting publication 
Chart showing the form of archaeobotanical data shared by year in primary archaeobotanical data publications 
The forms in which data are made available vary across journals  The sharing of primary data within an article remains the most common data sharing form in archaeobotany (Figure 1)  Data tables in text require manual handling to extract data, in journals such as VHA, whilst in other journals in-text tables can be downloaded as  csv files  These however would not be citable as a separate dataset  Supplementary datasets are the third most common form of data sharing  Indeed, the use of electronic supplementary material has been advocated recently for by some journals, such as the Journal of Archaeological Science (Torrence, Martinón-Torres & Rehren 2015)  Microsoft Excel spreadsheets are the most common form of supplementary data, followed by  pdfs and then word documents (Figure 1)  Both  xlsx and  docx are proprietary file formats, and not recommended for long term archiving or open science principles  There is no indication of improvement over the last decade in the form of data sharing  In 2018, 50% of articles did not share their primary data, and where the data was shared, it was in proprietary forms ( docx,  xlsx) or those that do not easily facilitate data reuse ( pdf) (Figure 3) 
Chart showing the location of archaeobotanical data from 2009–2018 in primary archaeobotanical data publications 
Just one of the articles included in this review incorporated a dataset archived in a repository (Farahani 2018), in contrast to the substantial growth in data repositories across academic disciplines (Marcial & Hemminger 2010)  Other examples provide the underlying data for monograph publications, such as that of the archaeobotanical data from Gordion, Turkey (Marston 2017a, 2017b), Silchester, UK (Lodwick 2018; University of Reading 2018) and Vaihingen, Germany (Bogaard 2011a; Bogaard, 2011b) 
Several of the journals that have been assessed have research data policies  In the case of Vegetation History and Archaeobotany, sufficient papers have been surveyed to assess the impact of the research data policy on the availability of data  Figure 4 show the proportion of data sharing formats through time just for VHA (note the small sample size)  The introduction of a research data policy in 2016 encouraging data sharing in repositories has not resulted in any datasets being shared in that format  Of the 10 articles published in PLOS One after the introduction of a clear research data policy in 2014, 4 did not contain primary data  However, elsewhere, journals with no research data policy, such as Antiquity, has one of the lower levels of data sharing (Figure 1) 
Chart showing the location of primary archaeobotanical data in Vegetation History and Archaeobotany 
There are various reasons for why a primary dataset may be lacking  The option of providing supplementary datasets has been available in many of the journals here since before the start of the surveyed period (e g  Vegetation History and Archaeobotany in 2004), and so cannot be a reason for the absence of data publication in this journal while it may be a reason in other journals  Reasons suggested for a lack of data sharing within archaeology include technological limitations, and resistance amongst some archaeologists to making their data available due to cautions of exposing data to scrutiny, lost opportunities of analysis before others use it and loss of ‘capital’ of data (Moore & Richards 2015: 34–35)  Furthermore, control over how data tables is presented (taxa ordering, summary data presented) may also contribute to the preferential publishing of data within journal articles  Another factor to consider is the emphasis on the creation of new data through archaeological research (Huvila 2016)  The creation of a new archaeobotanical dataset through primary analysis is a key form of training in archaeobotany, and the perception of the value of the reuse of other previously published archaeobotanical journals may be low, hence not encouraging the sharing of well-documented datasets  Excellent exams of data reuse have resulted in influential studies (Bogaard 2004; Riehl 2008; Wallace et al  2019), and would hopefully encourage further data sharing in the future 
Given that there are numerous examples of meta-analysis which do take place in archaeobotany, it seems likely that the prevalent form of data sharing is through informal data sharing between individual specialists  However, this does not improve access to data in the long term, and is inefficient and time consuming, with large potential for data errors (Kansa & Kansa 2013), and relies on personal networks, which are likely to exclude some researchers  The absence of primary data in many archaeobotanical publications thus inhibits the verification of patterns observed within a dataset, and strongly limits the re-use potential of a dataset 
4 2  Data citation
One of the common arguments for increasing data sharing is an associated increase in the citation of the articles which have data available  Here, the data citation practices of meta-analyses of plant macrofossil data undertaken over the last decade have been reviewed  20 journals were consulted, including a wider range of period-specific journals, and 107 articles were assessed (Lodwick 2019 Table 2)  Data citation was assessed as ‘in text’ or ‘in table’ to refer to when the citation and the bibliographic reference were within the article, as ‘in supplementary data’ when the citation and reference were within the supplementary materials, and as ‘no citation’ when no citation and reference was provided 
21% of articles (n = 22) did not contain any citations to the underlying studies  16% (n = 17) contained citations within supplementary data files  50% of articles (n = 53) contained a citation within a table within the main article, and 14% (n = 15) contained citations within the main text  For the 21% of articles without data citations, the results of these studies could not be reproduced without consulting individual authors  The papers supplying the underlying data also received no credit for producing these datasets  Where articles contain citations within the main article (in text or table), full credit is provided to the underlying studies, a citation link is created through systems such as google scholar, and the study can be easily built upon in the future  Where the citation is provided within supplementary data, the original studies do receive attribution, but are not linked to so easily 
Through time, there is a steady decrease in the proportion of studies without citations to the underlying data, whereby of the 17 meta-analysis articles published in 2018, only one had no data citations  In comparison, in 2009, 3 out of 8 meta-analysis articles contained no data citation (Figure 6)  Overall this is a more positive outlook on the reuse of published data, but the consistent presence of articles lacking data citation indicates that improvements are needed  Reasons for a lack of data citation may include restrictions on word counts imposed by journals, a lack of technical knowledge in making large databases available, or the wish to hold on to a dataset to optimise usage  Considering the type of journal (Figure 5), levels of data citation are worse in general archaeology journals, with sub-disciplinary journals showing slightly better levels of data citation  In particular VHA has a lack of consistency in where data citations are located 
Chart showing the location of data citations in meta-analysis journal articles by journal type 
Chart showing the location of data citations in meta-analysis journal articles from 2009–2018 
4 3  Reuse of archived archaeobotanical datasets
The majority of data citations assessed in the previous section are to articles or book chapters rather than data-sets  The ADS currently hosts 66 data archives which have been tagged as containing plant macro data, deriving mainly from developer-funded excavations but also some research excavations  However, in some of these the plant macro data is contained within a pdf  As, the archiving of archaeobotanical datasets in data repositories is still at an early stage, the reuse of these datasets is assessed here on a case-by-case basis  The archaeobotanical dataset from the Neolithic site of Vaihingen, Germany (Bogaard 2011b) has not been cited on google scholar  Metrics are provided through the ADS, showing this dataset has been downloaded 56 times with 477 individual visits (as of 25/2/19)  The archaeobotanical dataset from Gordion by Marston has no citations on Google Scholar (Marston 2017b), neither does the Giza botanical database (Malleson & Miracle 2018), but these are both very recently archived datasets  In contrast, the Roman Rural Settlement Project dataset, which includes site-level archaeobotanical data, has received greater levels of use, with 12 citations in Google Scholar, over 40,000 file downloads, and over 35,000 visits (Allen et al  2018) and the archaeobotanical computer database (Tomlinson & Hall 1996) has been cited 44 times, and is the major dataset underpinning other highly-cited studies (Van der Veen, Livarda & Hill 2008; Van der Veen, Hill & Livarda 2013)  Whilst there is clearly precedence for the reuse of archaeobotanical databases, current data citation practices within archaeobotany do not yet appear to be formally citing individual datasets, meaning an assessment of the reuse of archived archaeobotanical datasets is challenging 
5  Steps Forward
This review of data sharing, citation, and reuse practices in archaeobotany has found medium levels of data sharing, good levels of data citation, but so far limited levels of reuse of archived data sets  This picture is similar across archaeology, in part attributed to the status of archaeology as a small-science, where data-sharing takes place ad-hoc (Marwick & Pilaar Birch 2018)  Here, recommendations are discussed for improving these data practices within archaeobotany, of applicability more widely in archaeology 
Clearly an important step is improving the sharing of plant macrofossil data  Given the reasonable small size of most archaeobotanical datasets (a  csv file < 1mb), and a lack of ethical conflicts, there seems to be few reasons why the majority of archaeobotanical data couldn’t be shared  In the case of developer-funded derived data, issues of commercial confidentiality could limit the sharing of data  A key stage is establishing why levels of data sharing are not higher  Issues within archaeobotany may include the conflict between having to publish results within excavation monographs, which may take some time to be published, and have limited visibility due to high purchase costs and no digital access, and the need to publish journal articles for career progression within academia  The production of an archaeobotanical dataset is very time-consuming, and interim publication on notable aspects of an assemblage may be considered as a necessary publication strategy  More broadly, one important aspect is issues of equity in access to digital archiving resources (Wright & Richards 2018), such as differential access to funds, training and knowledge  A recent study in Sweden found that we need to know concerns, needs, and wishes of archaeologists in order to improve preservation of archaeological data (Huvila 2016), especially when control of ones data may be linked to perceptions of job security  In order to make improvements in data sharing and reuse across archaeology, we need improved training in data sharing and the reuse of data in higher education (Touchon & McCoy 2016; Cook et al  2018), improved training in data management (Faniel et al  2018), and crucially, the necessary software skills to make the reuse of archived datasets attainable (Kansa & Kansa 2014: 91)  Examples of good practice in archaeobotany are the Vaihingen and Gordion datasets which demonstrate how datasets can be archived in data repositories to accompany a monograph (Bogaard 2011b; Marston 2017b), whilst Farahani (2018) provides an excellent example of a journal article, where the primary data is supplied as a  csv in a cited data repository along with the R script for the analysis 
In tandem with the need to encourage authors to share their data, is the need for journals to create and implement research data policies  Given the existence of research data policies in many of the journals included here, this reflects other findings of the poor enforcement of data policies by journals (Marwick & Pilaar Birch 2018), supporting arguments that journals should not be relied upon to make data accessible, and data should instead by deposited in digital repositries  In order to implement change in data sharing, there is a role to play for learned societies and academic organisation in lobbying funding bodies, prioritising data sharing in research projects  A key step is through journal editorial boards, and the enforcement of any pre-existing research data policies (Nosek et al  2015)  Revi
Context 3:  We don't want someone else to make all these decisions for anybody  We don't even want the majority to rule  In every way that is possible, we want the minorities to control their own conditions in their own lives  There are very few things that are the province of government, but way too many things nowadays are being given to the government carelessly, fearfully, whatever  In my terms -- and I happen to be a right-wing person in terms of the economy and private freedoms -- I want more markets and fewer governments  Markets give choices to individuals  They let people trade what they don't want for what they do want  Again, to the extent possible, they want people to make individual choices 
What worries me is large concentrations of power, making choices for people  Big business, big government, even big media  The media until now have mostly been our protectors, because they go out and produce information, they use anonymous sources where necessary, and they make that information free  What protected global networking is going to do is give more and more of that power to individuals, and help reduce the power of big institutions of any kind  We are going to have small businesses flourishing, because it is easier for them to collect resources  You don't need to have a giant monolithic corporation to be efficient any more, and so a lot of marketplace economies of scale will even disappear, as we have better networking, better coordination  We have markets like the American Information Exchange, and if you don't know what that is, come and see me, or Hugh Daniel, or a couple of other people 
On the social side, I think 20 years ago    when you mentioned 15 years ago, I thought, Yes, that must have been about 1940  Then I realized    Anyway, some time ago there was all this talk about the global village  We're going to have mass broadcasting, we're going to have mass E-mail, we're going to have this global village  We don't  What we have is a lot of global villages, but as Mitch said, they're no longer geographical, physical villages  They're small, geographical villages of people with like interests  The big question becomes, How do we avert tribalism? It might not be nation against nation any more, but it certainly will be rich against poor, and franchised versus disenfranchised 
LIASSON: Thank you all very much  Now we can all try to stir up the pot a little bit  Somewhere between Mitch's paradise and the Simon's apocalypse is probably what's really going to happen  I want to just jump off from what Esther said about you all being in a minority and what kind of responsibility you owe to the rest of the world  We're in the midst of a presidential election and not one single candidate has said anything about Cyberspace  I am wondering if you think they should, and what are the kinds of extremely important issues that you think should be discussed? Should they be discussed in a kind of mass, political forum? Or should they be left to an elite like you to discuss and decide, and not really spend a whole lot of energy trying to translate or disseminate them to the great masses of people? I guess what I am wondering is, if you were an advisor to one of the presidential candidates, or a candidate yourself, how would you go about interjecting these things? Or wouldn't you bother at all?
DYSON: Does he want to get elected, or does he want to make a point?
LIASSON: I think he wants to make a point  If he wants to get elected, I think the discussion would stop right now 
DYSON: Let me just try a serious answer  I think what a candidate could say is, "I'm no longer going to protect the textile industry, the peanut butter interests, the sugar guys, the antediluvian steel mills  If I'm going to have an industrial policy and help anyone, it's going to be new technology  I'm going to focus on investment in R&D  I am going to create a national infrastructure for telecommunications, just the way we created a highway system years ago  I'm going to put people to work doing these things " I think that would go over reasonably well  I think it's something most of us would agree on  (laughter) We have an industrial policy -- we might as well acknowledge it, and we might as well have it be forward-looking 
KAPOR: Now there is something about the question as to whether this is presidential material that I think is ironic, given that most people really want to vote for "none of the above " We know in our hearts that we have come to a particular period in history in which the presidential spectacle seems to be particularly irrelevant to whatever set of problems we have on our minds  As a great believer in democracy, I think this is incredibly lamentable  We need to do something about this, because there are a lot of issues, but Cyberspace is not ready for prime time  It would be trivialized -- I have seen what Geraldo did to hackers, and I don't need to see any more 
It seems to me that the presidential candidates are really not the leaders that they ought to be, but are always putting their finger to the wind to see if they can detect some current of values or beliefs that can help get them elected  And I think that -- I'm not espousing utopian vision -- there needs to be an utopian vision out there, so people have something to give them some inspiration  But values are a lot more important than technology  There are some values in this community -- and I'm not sure if it's an elite or a minority or both -- but it's really in the propagation of a sense of values about openness and tolerance, acting on that basis and living one's life, and saving capitalism from itself and things like that where we can make a difference  If some of the expressions are technological, that's fine  We are living in an era where people like buttons, and so on  If we do that well, the presidential candidates are going to be coming to us 
LIASSON: You talk about Cyberspace not being ready for prime time -- I still want a definition of Cyberspace in 25 words or less -- but I think you want to transform prime time to a certain extent 
DYSON: Mostly I agree with this, but the press does have two roles: one is collecting information and uncovering things, and the other is setting the agenda  If 12,000 voices are crying out, who's going to listen to them? Who's going to notice when they do discover that the President did something wrong? Again, it's a check and balance sort of thing, but there is a certain community that is created by collective media 
KAPOR: Esther, what makes you believe that in Cyberspace Mara won't have two hours a day of her own that everyone listens to  (laughter) She might get more time than she gets today, because people trust her 
DYSON: But then she becomes prime time 
LIASSON: But you said before that instead of one global village, we have a lot of little global villages  I'm wondering if instead, we won't have millions of little huts  I mean individual huts  There are just so many different choices 
LIASSON: What I'm wondering is, if everybody becomes their own producer, publisher, what does that mean for the future?
KAPOR: I think we'll get a much more fluid, self-organizing state  I don't think in practice everybody is going to be what we think of today as a broadcast publisher  I just want things to be able to sort themselves out in a much more equitable fashion  We have this enormous, artificial scarcity today over the means of communication, because the government awards licenses which self-perpetuate  They are about to do the same thing, and give every broadcast television station another license for HDTV  So if you've got a license today, you get a second one; if you don't have one, you get nothing  That is going to be our policy about HDTV  I think it would be a lot better if we had more markets, more choices, and better values  I don't know how to do better values, but we know how to do more choices  So the point is, we'll wind up with some new regime which I don't think that we can particularly predict  I don't think that it is going to be chaotic or anarchic  I think there is something about people as social animals or creatures -- we will create some new forms of social organization  There will be information middlemen; there will be the equivalent of editors and packagers  There will be trusted intermediaries who help organize these new media  If you open it up and equalize things so that everybody can participate, you will get more diversity of points of view, you will get less homogenization  One of the reasons that tons of people have just dropped out, or are in terminal couch-potato-dom is that the sets of choices and the values that come across the tube are not ones that stir the human heart  And people know that  They can't figure out what to do about that, so they sort of fuzz out on drugs and alcohol  I say let's edit TV, which is the electronic drug  Let's do something about that 
DAVIES: I like your idea, Mitch  I think it's sweet  (laughter) The problem is that I really worry that the ultimate test of the future is going to be the outcome of the quest, the battle between those who are looking for the sort of vision you've got of the right of the individual, the individual being the producer  And that, probably, is the way we solve our problems on this planet  But there is the other side, and that's the planetary managers  Planetary management is the path of the least resistance  You know all the powermongers go for the planetary management model, because they all think they can clamber over the bodies to get to the top  Ultimately the test is going to be who comes out on the top, the individual rightist or the planetary managers  Unfortunately, I'm not a betting man, but at the moment I'd like to bet on the planetary managers 
DYSON: Part of this issue is reducing the value of incumbency, whether it's incumbency in prime time live, or incumbency in the government  There is much more fluidity of movement; you can't accumulate power because the unorganized forces have more power than you do 
P  DENNING: I feel a little strange being on the left end of the stage, because most people think of me as being on the far right sometimes, but right now I'd like to comment on something that is halfway between what Mitch is saying, and what Simon is saying  The way I hear what Simon is saying, is that there is a disease of today which I will call inward- centeredness  We are very worried about ourselves and our organizations  We find in that orientation a lot of instability of things and technologies that change rapidly  In order to achieve the world that Mitch is talking about, we need to cure the disease, and instead come from an orientation that we could call outward-centeredness, instead of inward-centeredness  The question is the shift from, How do we accumulate power? to, How do we help others accumulate power? How do we go from looking for stability in things to looking for stability in relationships? In watching my own children grow up, I am convinced that they know more about this than I do  In listening to some of the younger people here, I'm more convinced that they know more about this than I do  They know something about the outward-centeredness that I have yet to learn  Observing this among children and among students gives me a lot of optimism, as a matter of fact, against the apocalypse that Simon talks about, because Simon is talking about the world that would be created if we continued "us," and I think that the world that is being created by our children with their outward-centeredness is going to be the kind of world that Mitch is pointing towards  And I am much more optimistic about that than Simon is 
LIASSON: Roland, I wonder if we can interject you into this discussion a little bit  You have been a policymaker  What can be done to make sure that Simon's vision doesn't come true, and something a little closer to what Esther and Mitch describe does happen?
HOMET: I think we probably need both doom seers and paradise seekers  We'll always have them, and we should have them  It's between the swing of those two views that things happen  I think that this notion of replacing the gatekeepers and letting everybody perform his own dance, to the amusement of those who chose to tune in, is one that many of us were promoting 20 years ago  That's not 1940 -- that's 1970 (laughter), and we were quite convinced that was likely to happen by the end of that decade  Now it's 12 years beyond the end of that decade, and we're nowhere near having that happening  We just have newly-named controversies, and so, as you heard me say in my little short remark, I think that our objective ought to be more modest, and that is to keep the questions open, not let them be foreclosed -- certainly not prematurely, and not on the basis of inadequate evidence  I would say something about the apocalyptic view, which is, I think there is a difference between information policy questions and welfare questions  The poor we have always with us, as somebody once said, and whether information, Cyberspace -- whatever you want to call it -- is promoted or not, that is true  It may become more glaringly true in an advanced information society, in which case, more may be done about it  So I wouldn't despair about that, and I wouldn't hold back on the development of instruments of interconnection simply because we can see that there is and will remain an underclass  Perhaps if we do the one, we'll be better equipped to do the other 
LIASSON: In just a minute or two, we're going to open this up to your questions, but I want to try to end maybe with a discussion of something quite specific, which is, Who should own the new infrastructure and information systems? Should they be publicly owned? There are lots of conflicts even within the vision that you lay out 
KAPOR: The first point I'd make is let's not make the unnecessary mistake of betting on a single infrastructure  Technologically, we don't need to do that  In the 1930s, pre-digital, the old Bell system was the social contract  You get a monopoly, you have an obligation to provide universal service  We've learned a few things about how to do things with interoperable standards and how to interconnect multiple, independent providers and carriers  One of the fathers of the Internet, Vint Cerf, is sitting here in the front row, and he deserves an enormous amount of credit for insisting on this vision and promulgating it  A lot of the risks that come with private ownership of infrastructure go away when it's no longer a monopoly  The abusive problems that are sometimes experienced with local phone service and cable companies -- both of which are private sector monopolies -- I would say come more from not their private sector character, but from their monopoly character  If it is possible for there to be competition, that serves as the most effective check that we know of in this society against abuse  So I would opt for private infrastructure, but lots of it  Government has to make sure that everybody stays interconnected -- it's the referee that keeps the playing field level, doesn't let people cheat, and sort of bangs a few heads together when people get a little too greedy, or a little too selfish  If we do that, that will provide for the most choice and the most diversity 
LIASSON: Are we all in agreement on that?
HOMET: Not entirely  I think the question is less who should own infrastructure than how it should be classified  There may be a role for government in, for example, extending communication pipes to rural America for at least a period, as with the TVA  We have always had that question  There has always been a mixed economy with government doing some things and private sector others  It's a debate and should be a debate about who does what best



ground_truth: Technological limitations, resistance to exposing data to scrutiny, and desire to hold onto data for personal use.

prompt_front: Now, answer the following question based on the text below, only give me the answer and do not output any other words.

Question: Who is the program chair of this conference?

Read the following text and answer briefly.

Context 3:  It should be revised from time to time, but the important question is, If we get a significant distribution system like cable television, how should we classify it? I speak here from the heart, because 20 years ago, I was trying to fasten onto, or gain the recognition for, cable as a broadband distribution system which was only trivially in the program production and publishing business, but was very much in the distribution business and ought to have been treated as a common carrier open to all information suppliers  Had that happened, we would have been very much further along in the vision that some of us had 20 years ago  (applause) It tends to support what I said about not going in for premature freezing or characterization of how things look  It was decided, because the broadcasters felt threatened, to treat cable as a species of broadcasting  That's the greatest frittering away of resources in my lifetime, and perhaps in the lifetime of the United States of America  Let's not make that mistake again  Let's be clear-eyed and ask the broad-scale questions about public use and benefit  Thank you 
LIASSON: Let's open it up to the audience  If you have any questions     oh my God, wrestle your way to the microphone!
AUDIENCE MEMBER: Let us not forget the history of the commons in which a wealthy society creates in its overflowing abundance structures on which all people can participate  This was originally, back in medieval society, the structure that was created for the support of the poor  In the abundance of the land in which the overpopulation was not a question, and there was much agriculture to go around, and the poor were supported out of the commonly-owned things that were jointly owned by all society  That's all I have to say 
LIASSON: Who wants to start?
DAVIES: Sticking to my apocalyptic vision just for the moment, because that's how I'm characterized, what I would like to see, just as my own social experiment, if you like, is for the various groups that this room represents and groups that you are all involved in, is to actually set up the apocalyptic vision, and then see how you as part of the information technology community can utilize it, stop it, or reverse it  It's only when you see the vision and see your own part in it that we are actually going to set up solutions  I mean, that is a straight, outright homework assignment, and I think would be a great benefit for everybody  Then go on and publish them through the E-mail, or the Internet, whatever 
DYSON: Something along the lines of go find the most influential person you know well enough to influence, who you do not agree with -- assuming that you all agree with me, of course -- and attempt to win that person over to your point of view  In other words, don't stick to your own community  Don't just talk to the people who only agree with you  Go out and evangelize or proselytize to people who don't understand what this stuff is about  Do it in such a way that you are not superior or offputting; don't try to be right; try to win and expand this community, not in terms of pressure or rightness, but in terms of understanding what we are about  The biggest problem is ganging up on some of these politicians and having them think that this stuff is not cute, or weird, or colorful, or irrelevant, but incredibly important  Make the rest of the world know about us 
HOMET: I would like to second that motion  The story is told that when a beautiful woman comes out on a street in Paris, every man within eyeshot becomes in that instant much more intensively himself  (laughter) What I would suggest to you, if you are energized by this subject, is to be yourself  To thine own self be true, and perhaps to add to that the biblical admonition to the apostles -- if I remember it correctly -- and this picks up what Esther was saying -- to be wise as snakes, and cunning as foxes  Go out there to persuade 
P  DENNING: I'd like to add to that  It is not only within yourself that you have to look, it's within others  Don't assume that you know the answers, but go talk to people  Don't just talk to us, because we already know what "us" has to say, but go to talk to people that we haven't talked to and find out what concerns them 
AUDIENCE MEMBER: Hi, my name is Lou Woleneck  I'm from the LBJ School of Public Affairs at the University of Texas  I'm a graduate student  I have a question, a general policy question, about how we should go about providing the information resources to the have-nots that the information elites have access to now  What sort of strategy that you all would have for that?
KAPOR: A 30-second or less answer, which is to set a national policy that updates a universal service for the 21st century that says everybody needs to have basic minimal access to a digital platform that reaches into every home, into every office and school in the country  We should focus our attention on how to put in place the least expensive amount of infrastructure that will produce that  What we find is, if we do that, then the overwhelming majority of American families will find it already within their budget to be able to do that, because it will be priced like basic phone service  To the extent that we need to continue or even slightly expand the kinds of lifeline programs that subsidize today's basic voice telephone service for a small percentage of the population, we should be prepared to renew that commitment  We don't need to bankrupt ourselves to give everybody access to a digital platform 
JIM WARREN: My name is Jim Warren  Two quick observations: there were several cynical comments during the last several days about a number of IRS people being here  It turns out, because they never had a platform to say this, that the whole crowd from the IRS who are here, as I understand it, are from the IRS privacy project, intent on developing policies to assure privacy protection for taxpayer information  So let us not be so cynical about their being here; otherwise, remember that they are simply doing what they are told to do by our representatives  (laughter and hisses) I was also bothered by both Simon's, and (my God!) Esther's comments on those evil little men, and the men in politics, etc  Gee, this is a modern age, let's say "men and women," for evil deeds, as well as good deeds 
DYSON: There aren't enough women in politics for there to be any evil ones 
WARREN: Well, I am sure that I can find some evil ones for you  (laughter) Anyway, to the main points: I would say that we are not so much elite, in that we are open to anyone who takes the initiative to join us, and many of us are active mentors in trying to get others to join us  I would say simply that we are a minority, and it occurs to me that revolution has always been a minority activity  It was not millions of Russians who opposed the attempted coup several months ago  It was ten, twenty, or thirty thousand in Moscow, with the aid of communications  It was not a massive movement, a populist movement, in America that resisted the Crown, two centuries ago  It was a small minority of activists and we are the activists here -- we are the revolutionaries  Freedom has always been a do-it-yourself activity, but the key syllable in that word activity is act  Let us reaffirm freedom of speech, press, assembly, security against undue search and seizure -- the basic constitutional freedoms and privileges  Let us demand that our politicians and our political candidates do the same in explicit formal commitments to act in behalf of protecting electronic civil liberties, just as they validate and speak favorably for traditional civil liberties  We can write our politicians, write our candidates and say, "Take a position in favor of civil liberties, regardless of the technology of the moment " Thank you 
GLENN TENNEY: Thank you for the introduction, Jim 
LIASSON: Are you from the IRS?
TENNEY: No  (laughter) My name is Glenn Tenney, and I have a question for you, Mara  I think that I have enough supporters on the panel  I'm not too curious about their views, but they are welcome to them  You questioned if the presidential election and race is ready for Cyberspace  What about Congress? I'm running for Congress -- is it ready for me?
AUDIENCE MEMBER: Ms  Liasson, I believe that you have opened a can of worms called politics for this little hacker community  You certainly have with me in your comment about asking for comments for the Cyberspace era from presidential candidates  I have very strong reactions to that  I think that I am going to try to express them, as a pure statement, or maybe an actual story  Several years ago, I was discussing with a friend of mine the current presidential, the then-current presidential election  He was asking me why I wasn't rabidly supporting Jesse Jackson  I thought about it, and my first response was, "Well, let's talk about the other candidates for a second  What about -- and I'll take a random name -- Michael Dukakis?" And my friend looked at me and said, "Michael Dukakis, he's just an administrator, he's not a visionary " I thought about it, and I said, "Hold on, I'm an American, I'm not someone who's a slave of the Queen of England, or something like that  I'm my own visionary, I decide where I am going " I don't want the politicians walking around telling me that I am going to have an expressway system that's going to pave over all my favorite swamps to play in  I don't want the politicians walking around defining what I'm going to do in my life  I want to elect politicians to manage government for me, to provide the barest minimum necessities to keep us smoothly greased as individuals in living together, and I want those politicians to be of the people, and I don't want them to tell me what my opinions should be  Finally, I want to cap that off with when we have government deciding how our systems work for us, we can then end up with situations where we can say, "Oh yeah, that IRS guy or that government net guy, he was just doing his job when he banned cryptography," or something like that  That's not the sort of world that I want to live in  I want to live in a world, where each of us defines our little space in it  Thank you all 
LIASSON: I think we have time for just two more and then we'll have to wrap it up 
AUDIENCE MEMBER: Hi, to the apocalypse types  I'd like to say just one thing that somebody said: The truth will make you free  In that this technology is a vehicle of communication, I believe that it is a vehicle of the truth, and as long as we keep it free, the truth will be heard that much more  Now I have kind of a question with a bit of a statement  I am a learning-disabled college student  I didn't ever finish high school  I had a freshmen education in high school, because of educational problems, and adjustment problems, I never really got too far beyond that  I write probably a fifth of the speed of anyone in this room and I have a real hard time doing math without a calculator  That's part of the reason why I wasn't able to do well in school  I read very well, fortunately, so I was able to go in when I was eighteen and take my GED just flat out without studying for it  I'm not dumb, or uneducated by any standards, but what has allowed me to get an associate's degree in college, and what has allowed me to approach graduation and get a bachelor's degree in college is the kind of technology that we are dealing with  I have never had easy access to that technology  The barriers that I have faced have been ones of order, regimentation, and where people try and say, "Oh well, you don't fit in, you're not a CS student, you don't need those resources " I'm good with computers, I do a lot with them, I spend a lot of time with them  I hack, I don't do anything illegal, but I took a hacksaw to the frame of my nasty little 8088 about two years ago to cram some RAM into it, because that was the only way I could get it to fit and I needed it  Now I'm in a little bit better shape  I'm approaching the point where I would like to see ISDN real soon, because I need that kind of connectivity  You know, I'm doing interesting things that I find absolutely wonderful, but the idea that the kind of technology that is available to us, that is just there for the using, could be limited and unavailable to people, or that people would have to go through some of the things that I have had to go through, not being able to do well on tests, because I had no word processor available to me  That type of thing, even though they are all over the place, elsewhere  It was just that that wasn't an acceptable solution  That type of policy planning, that type of government, that type of order scares me  And I have to ask, what is your answer to that?
DAVIES: The apocalyptic vision of a world in grief and individual rights in crisis has nothing to do with a Luddite mentality, and it would be very dangerous for the people in this room to link the two together  I, for one, believe in technology  I am very grateful for it, and I think the world is a better place for it  I have great faith in the future, but technology's not a silver lining for the future  It's not an El Dorado, it's more like plutonium  The very great thing that technology does for all of us can also be used by the people who would repress our freedoms and all I am saying is be aware of that  Let's not marginalize people like me, who are saying, Hey look, we are going to have 15 billion people on the planet  We are going to have a political inversion, you know, that is going to create massive tensions that are going to repress our rights, or at least create a tension that we have never known before  Don't marginalize me -- don't shoot the messenger  I believe in technology, so please don't equate the apocalypse with Ludditism -- the two do not match 
LIASSON: We're about out of time  I'm going to turn this over to Lance 
HOFFMAN: Thank you, Mara  I'm really unhappy that we are out of time, but I feel that we have a contract to those who want to leave in a moment or two  Those who want to stay, can stay up here, are welcome to continue, until the hotel throws us out  Since Lu Kleppinger is in the room at the moment, I don't know when that will be, but we can probably have it for a little while  I just want to make a couple of comments before I formally close this meeting 
We have seen an awful lot happen in these last three days and there has been building, and indeed we will be continuing to some extent the work that Jim Warren started at CFP-1 -- a sense of community  It has been increased by the participation of various diverse groups  My one hope is that you do not stop that here  When each and every one of you goes home, contact -- I don't care whether it's by letter, or electronic mail, or even telephone, if you must -- three people that you have met here that you didn't know, or didn't know very well before, or perhaps only knew electronically, and now you know them in person, and continue talking with them and to their friends and colleagues  If you do that, this will be a success 
The other comment that I want to make is that Bruce Koball is going to need a lot of help for CFP-3  Please talk to him -- he is listed in the roster  Or better yet, don't do that, talk to him here, and then give him a month to chill out in Berkeley before he has to start working real hard  Check the message board, there are some messages that have not been picked up  You have your evaluation forms  If you haven't filled them out and you would like to, please do and turn them in  I have nothing else, except to thank you all for being such a good group and, hopefully, we'll see you next year in California  Thank you very much 
Support efforts at engaging society and government on the appropriate legal and social uses of technology 



ground_truth: Peter Denning.