Test Bank for Principles of Development, 6th Edition, 6e by Lewis Wolpert, Cheryll Tickle, and Alfonso Martinez Arias TEST BANK ISBN-13: 9780198800569 FULL CHAPTERS INCLUDED 1 History and basic... concepts The origins of developmental biology 1.1 Aristotle first defined the problem of epigenesis versus preformation 1.2 Cell theory changed how people thought about embryonic development and heredity 1.3 Two main types of development were originally proposed 1.4 The discovery of induction showed that one group of cells could determine the development of neighboring cells 1.5 Developmental biology emerged from the coming together of genetics and embryology 1.6 Development is studied mainly through selected model organisms 1.7 The first developmental genes were identified as spontaneous mutations Summary A conceptual tool kit 1.8 Development involves the emergence of pattern, change in form, cell differentiation, and growth 1.9 Cell behavior provides the link between gene action and developmental processes 1.10 Genes control cell behavior by specifying which proteins are made 1.11 The expression of developmental genes is under tight control 1.12 Development is progressive and the fates of cells become determined at different times 1.13 Inductive interactions make cells different from each other 1.14 The response to inductive signals depends on the state of the cell 1.15 Patterning can involve the interpretation of positional information 1.16 Lateral inhibition can generate spacing patterns 1.17 Localization of cytoplasmic determinants and asymmetric cell division can make daughter cells different from each other 1.18 The embryo contains a generative rather than a descriptive program 1.19 The reliability of development is achieved by various means 1.20 The complexity of embryonic development is due to the complexity of cells themselves 1.21 Development is a central element in evolution Summary Summary to Chapter 1 2 Development of the Drosophila body plan Drosophila life cycle and overall development 2.1 The early Drosophila embryo is a multinucleate syncytium 2.2 Cellularization is followed by gastrulation and segmentation 2.3 After hatching, the Drosophila larva develops through several larval stages, pupates, and then undergoes metamorphosis to become an adult 2.4 Many developmental genes were identified in Drosophila through large-scale genetic screening for induced mutations Summary Setting up the body axes 2.5 The body axes are set up while the Drosophila embryo is still a syncytium 2.7 Three classes of maternal genes specify the antero-posterior axis 2.8 Bicoid protein provides an antero-posterior gradient of a morphogen 2.9 The posterior pattern is controlled by the gradients of Nanos and Caudal proteins 2.10 The anterior and posterior extremities of the embryo are specified by activation of a cell-surface receptor 2.11 The dorso-ventral polarity of the embryo is specified by localization of maternal proteins in the egg vitelline envelope 2.12 Positional information along the dorso-ventral axis is provided by the Dorsal protein Summary Localization of maternal determinants during oogenesis 2.13 The antero-posterior axis of the Drosophila egg is specified by signals from the preceding egg chamber and by interactions of the oocyte with follicle cells 2.14 Localization of maternal mRNAs to either end of the egg depends on the reorganization of the oocyte cytoskeleton 2.15 The dorso-ventral axis of the egg is specified by movement of the oocyte nucleus followed by signaling between oocyte and follicle cells Summary Patterning the early embryo 2.16 The expression of zygotic genes along the dorso-ventral axis is controlled by Dorsal protein 2.17 The Decapentaplegic protein acts as a morphogen to pattern the dorsal region 2.18 The antero-posterior axis is divided up into broad regions by gap gene expression 2.19 Bicoid protein provides a positional signal for the anterior expression of zygotic hunchback 2.20 The gradient in Hunchback protein activates and represses other gap genes Summary Activation of the pair-rule genes and the establishment of parasegments 2.21 Parasegments are delimited by expression of pair-rule genes in a periodic pattern 2.22 Gap gene activity positions stripes of pair-rule gene expression Summary Segmentation genes and segment patterning 2.23 Expression of the engrailed gene defines the boundary of a parasegment, which is also a boundary of cell lineage restriction 2.24 Segmentation genes stabilize parasegment boundaries 2.25 Signals generated at the parasegment boundary delimit and pattern the future segments Summary Specification of segment identity 2.26 Segment identity in Drosophila is specified by Hox genes 2.27 Homeotic selector genes of the bithorax complex are responsible for diversification of the posterior segments 2.28 The Antennapedia complex controls specification of anterior regions 2.29 The order of Hox gene expression corresponds to the order of genes along the chromosome 2.30 The Drosophila head region is specified by genes other than the Hox genes Summary Summary to Chapter 2 3 Vertebrate development I: life cycles and experimental techniques Vertebrate life cycles and outlines of development 3.1 The frog Xenopus laevis is the model amphibian for studying development of the body plan 3.2 The zebrafish embryo develops around a large mass of yolk 3.3 Birds and mammals resemble each other and differ from Xenopus in some important features of early development 3.4 The early chicken embryo develops as a flat disc of cells overlying a massive yolk 3.5 The mouse egg has no yolk and early development involves the allocation of cells to form the placenta and extra-embryonic membranes Experimental approaches to studying vertebrate development 3.6 Gene expression in embryos can be mapped by in situ nucleic acid hybridization 3.7 Fate mapping and lineage tracing reveal which cells in which parts of the early embryo give rise to particular adult structures 3.9 Developmental genes can be identified by spontaneous mutation and by large-scale mutagenesis screens 3.10 Transgenic techniques enable animals to be produced with mutations in specific genes 3.11 Gene function can also be tested by transient transgenesis and gene silencing Human embryonic development 3.12 The early development of a human embryo is similar to that of the mouse 3.13 The timing of formation and the anatomy of the human placenta differs from that in the mouse 3.14 Some studies of human development are possible but are subject to strict laws Summary to Chapter 3 4 Vertebrate development II: Xenopus and zebrafish Setting up the body axes 4.1 The animal–vegetal axis is maternally determined in Xenopus 4.2 Local activation of Wnt/β-catenin signaling specifies the future dorsal side of the embryo 4.3 Signaling centers develop on the dorsal side of the blastula Summary The origin and specification of the germ layers 4.4 The fate map of the Xenopus blastula makes clear the function of gastrulation 4.5 Cells of the early Xenopus embryo do not yet have their fates determined and regulation is possible 4.6 Endoderm and ectoderm are specified by maternal factors, whereas mesoderm is induced from ectoderm by signals from the vegetal region 4.7 Mesoderm induction occurs during a limited period in the blastula stage 4.8 Zygotic gene expression is turned on at the mid-blastula transition 4.9 Mesoderm-inducing and patterning signals are produced by the vegetal region, the organizer, and the ventral mesoderm 4.10 Members of the TGF-β family have been identified as mesoderm inducers ▪ Experimental Box 4D Investigating receptor function using dominant-negative proteins 4.11 The zygotic expression of mesoderm-inducing and patterning signals is activated by the combined actions of maternal VegT and Wnt signaling 4.12 Threshold responses to gradients of signaling proteins are likely to pattern the mesoderm Summary The Spemann organizer and neural induction ▪ Cell Biology Box 4E The fibroblast growth factor signaling pathway 4.13 Signals from the organizer pattern the mesoderm dorso-ventrally by antagonizing the effects of ventral signals 4.14 The antero-posterior axis of the embryo emerges during gastrulation 4.15 The neural plate is induced in the ectoderm 4.16 The nervous system is patterned along the antero-posterior axis by signals from the mesoderm 4.17 The final body plan emerges by the end of gastrulation and neurulation Summary Development of the body plan in zebrafish 4.18 The body axes in zebrafish are established by maternal determinants 4.19 The germ layers are specified in the zebrafish blastoderm by similar signals to those in Xenopus 4.20 The shield in zebrafish is the embryonic organizer Summary to Chapter 4 5 Vertebrate development III: chick and mouse—completing the body plan Development of the body plan in chick and mouse and generation of the spinal cord 5.1 The antero-posterior polarity of the chick blastoderm is related to the primitive streak 5.2 Early stages in mouse development establish separate cell lineages for the embryo and the extra-embryonic structures 5.4 The fate maps of vertebrate embryos are variations on a basic plan 5.5 Mesoderm induction and patterning in the chick and mouse occurs during primitive streak formation 5.6 The node that develops at the anterior end of the streak in chick and mouse embryos is equivalent to the Spemann organizer in Xenopus 5.7 Neural induction in chick and mouse is initiated by FGF signaling with inhibition of BMP signaling being required in a later step 5.8 Axial structures in chick and mouse are generated from self-renewing cell populations Summary Somite formation and antero-posterior patterning 5.9 Somites are formed in a well-defined order along the antero-posterior axis 5.10 Identity of somites along the antero-posterior axis is specified by Hox gene expression 5.11 Deletion or overexpression of Hox genes causes changes in axial patterning 5.12 Hox gene expression is activated in an anterior to posterior pattern 5.13 The fate of somite cells is determined by signals from the adjacent tissues Summary The origin and patterning of neural crest 5.14 Neural crest cells arise from the borders of the neural plate and migrate to give rise to a wide range of different cell types 5.15 Neural crest cells migrate from the hindbrain to populate the branchial arches Summary Determination of left–right asymmetry 5.16 The bilateral symmetry of the early embryo is broken to produce left–right asymmetry of internal organs 5.17 Left–right symmetry breaking may be initiated within cells of the early embryo Summary Summary to Chapter 5 6 Development of nematodes and sea urchins Nematodes 6.1 The cell lineage of Caenorhabditis elegans is largely invariant 6.2 The antero-posterior axis in Caenorhabditis elegans is determined by asymmetric cell division 6.3 The dorso-ventral axis in Caenorhabditis elegans is determined by cell–cell interactions 6.4 Both asymmetric divisions and cell–cell interactions specify cell fate in the early nematode embryo 6.5 Cell differentiation in the nematode is closely linked to the pattern of cell division 6.6 Hox genes specify positional identity along the antero-posterior axis in Caenorhabditis elegans 6.7 The timing of events in nematode development is under genetic control that involves microRNAs 6.8 Vulval development is initiated through the induction of a small number of cells by short-range signals from a single inducing cell Summary Echinoderms 6.9 The sea urchin embryo develops into a free-swimming larva 6.10 The sea urchin egg is polarized along the animal–vegetal axis 6.11 The sea urchin fate map is finely specified, yet considerable regulation is possible 6.12 The vegetal region of the sea urchin embryo acts as an organizer 6.13 The sea urchin vegetal region is demarcated by the nuclear accumulation of β-catenin 6.14 The animal–vegetal axis and the oral–aboral axis can be considered to correspond to the antero-posterior and dorso-ventral axes of other deuterostomes 6.15 The pluteus skeleton develops from the primary mesenchyme 6.16 The oral–aboral axis in sea urchins is related to the plane of the first cleavage 6.17 The oral ectoderm acts as an organizing region for the oral–aboral axis Summary Summary to Chapter 6 7 Morphogenesis: change in form in the early embryo Cell adhesion 7.1 Sorting out of dissociated cells demonstrates differences in cell adhesiveness in different tissues 7.2 Cadherins can provide adhesive specificity 7.3 The activity of the cytoskeleton regulates the mechanical properties of cells and their interactions with each other 7.4 Transitions of tissues from an epithelial to a mesenchymal state, and vice versa, involve changes in adhesive junctions Summary Cleavage and formation of the blastula 7.5 The orientation of the mitotic spindle determines the plane of cleavage at cell division 7.6 The positioning of the spindle within the cell also determines whether daughter cells will be the same or different sizes 7.7 Cells become polarized in the sea urchin blastula and the mouse morula 7.8 Fluid accumulation as a result of tight-junction formation and ion transport forms the blastocoel of the mammalian blastocyst Summary Gastrulation movements 7.9 Gastrulation in the sea urchin involves an epithelial-to-mesenchymal transition, cell migration, and invagination of the blastula wall 7.10 Mesoderm invagination in Drosophila is due to changes in cell shape controlled by genes that pattern the dorso-ventral axis 7.11 Germ-band extension in Drosophila involves myosin-dependent remodeling of cell junctions and cell intercalation 7.12 Planar cell polarity confers directionality on a tissue 7.13 Gastrulation in amphibians and fish involves involution, epiboly, and convergent extension 7.14 Xenopus notochord development illustrates the dependence of medio-lateral cell elongation and cell intercalation on a pre-existing antero-posterior polarity 7.15 Gastrulation in chick and mouse embryos involves the separation of individual cells from the epiblast and their ingression through the primitive streak Summary Neural tube formation 7.16 Neural tube formation is driven by changes in cell shape and convergent extension Summary Formation of tubes and branching morphogenesis 7.17 The Drosophila tracheal system is a prime example of branching morphogenesis 7.18 The vertebrate vascular system develops by vasculogenesis followed by sprouting angiogenesis 7.19 New blood vessels are formed from pre-existing vessels in angiogenesis Summary Cell migration 7.20 Embryonic neural crest gives rise to a wide range of different cell types 7.21 Neural crest migration is controlled by environmental cues 7.22 The formation of the lateral-line primordium in fishes is an example of collective cell migration 7.23 Body wall closure occurs in Drosophila, Caenorhabditis, mammals, and chick Summary Summary to Chapter 7 8 Cell differentiation and stem cells The control of gene expression 8.1 Control of transcription involves both general and tissue-specific transcriptional regulators 8.2 Gene expression is also controlled by epigenetic chemical modifications to DNA and histone proteins that alter chromatin structure 8.3 Patterns of gene activity can be inherited by persistence of gene-regulatory proteins or by maintenance of chromatin modifications 8.4 Changes in patterns of gene activity during differentiation can be triggered by extracellular signals Summary 8.5 Muscle differentiation is determined by the MyoD family of transcription factors 8.6 The differentiation of muscle cells involves withdrawal from the cell cycle, but is reversible 8.7 All blood cells are derived from multipotent stem cells 8.8 Intrinsic and extrinsic changes control differentiation of the hematopoietic lineages 8.9 Developmentally regulated globin gene expression is controlled by control regions far distant from the coding regions 8.10 The epidermis of adult mammalian skin is continually being replaced by derivatives of stem cells 8.11 Stem cells use different modes of division to maintain tissues 8.12 The lining of the gut is another epithelial tissue that requires continuous renewal 8.13 Skeletal muscle and neural cells can be renewed from stem cells in adults 8.14 Embryonic stem cells can proliferate and differentiate into many cell types in culture and contribute to normal development in vivo Summary The plasticity of the differentiated state 8.15 Nuclei of differentiated cells can support development 8.16 Patterns of gene activity in differentiated cells can be changed by cell fusion 8.17 The differentiated state of a cell can change by transdifferentiation 8.18 Adult differentiated cells can be reprogrammed to form pluripotent stem cells 8.19 Stem cells could be a key to regenerative medicine 8.20 Various approaches can be used to generate differentiated cells for cell-replacement therapies Summary Summary to Chapter 8 9 Germ cells, fertilization, and sex determination The development of germ cells 9.1 Germ cell fate is specified in some embryos by a distinct germplasm in the egg 9.2 In mammals germ cells are induced by cell–cell interactions during development 9.3 Germ cells migrate from their site of origin to the gonad 9.4 Germ cells are guided to their destination by chemical signals 9.5 Germ cell differentiation involves a halving of chromosome number by meiosis 9.6 Oocyte development can involve gene amplification and contributions from other cells 9.7 Factors in the cytoplasm maintain the totipotency of the egg 9.8 In mammals some genes controlling embryonic growth are ‘imprinted’ Summary Fertilization 9.9 Fertilization involves cell-surface interactions between egg and sperm 9.10 Changes in the egg plasma membrane and enveloping layers at fertilization block polyspermy 9.11 Sperm–egg fusion causes a calcium wave that results in egg activation Summary Determination of the sexual phenotype 9.12 The primary sex-determining gene in mammals is on the Y chromosome 9.13 Mammalian sexual phenotype is regulated by gonadal hormones 9.14 The primary sex-determining factor in Drosophila is the number of X chromosomes and is cell autonomous 9.15 Somatic sexual development in Caenorhabditis is determined by the number of X chromosomes 9.16 Determination of germ cell sex depends on both genetic constitution and intercellular signals 9.17 Various strategies are used for dosage compensation of X-linked genes Summary Summary to Chapter 9 10 Organogenesis The insect wing and leg 10.1 Imaginal discs arise from the ectoderm in the early Drosophila embryo 10.3 The adult wing emerges at metamorphosis after folding and evagination of the wing imaginal disc 10.4 A signaling center at the boundary between anterior and posterior compartments patterns the Drosophila wing disc along the antero-posterior axis 10.5 A signaling center at the boundary between dorsal and ventral compartments patterns the Drosophila wing along the dorso-ventral axis 10.6 Vestigial is a key regulator of wing development that acts to specify wing identity and control wing growth 10.7 The Drosophila wing disc is also patterned along the proximo-distal axis 10.8 The leg disc is patterned in a similar manner to the wing disc, except for the proximo-distal axis 10.9 Different imaginal discs can have the same positional values Summary The vertebrate limb 10.10 The vertebrate limb develops from a limb bud and its development illustrates general principles 10.11 Genes expressed in the lateral plate mesoderm are involved in specifying limb position, polarity, and identity 10.12 The apical ectodermal ridge is required for limb-bud outgrowth and the formation of structures along the proximo-distal axis of the limb 10.13 Formation and outgrowth of the limb bud involves oriented cell behavior 10.14 Positional value along the proximo-distal axis of the limb bud is specified by a combination of graded signaling and a timing mechanism 10.15 The polarizing region specifies position along the limb’s antero-posterior axis 10.16 Sonic hedgehog is the polarizing region morphogen 10.17 The dorso-ventral axis of the limb is controlled by the ectoderm 10.18 Development of the limb is integrated by interactions between signaling centers 10.19 Hox genes have multiple inputs into the patterning of the limbs 10.20 Self-organization may be involved in the development of the limb 10.21 Limb muscle is patterned by the connective tissue 10.22 The initial development of cartilage, muscles, and tendons is autonomous 10.23 Joint formation involves secreted signals and mechanical stimuli 10.24 Separation of the digits is the result of programmed cell death Summary Teeth 10.25 Tooth development involves epithelial–mesenchymal interactions and a homeobox gene code specifies tooth identity Summary Vertebrate lungs 10.26 The vertebrate lung develops from a bud of endoderm 10.27 Morphogenesis of the lung involves three modes of branching Summary The vertebrate heart 10.28 The development of the vertebrate heart involves morphogenesis and patterning of a mesodermal tube The vertebrate eye 10.29 Development of the vertebrate eye involves interactions between an extension of the forebrain and the ectoderm of the head Summary Summary to Chapter 10 11 Development of the nervous system Specification of cell identity in the nervous system 11.1 Initial regionalization of the vertebrate brain involves signals from local organizers 11.2 Local signaling centers pattern the brain along the antero-posterior axis 11.3 The cerebral cortex is patterned by signals from the anterior neural ridge 11.4 The hindbrain is segmented into rhombomeres by boundaries of cell-lineage restriction 11.5 Hox genes provide positional information in the developing hindbrain 11.6 The pattern of differentiation of cells along the dorso-ventral axis of the spinal cord depends on ventral and dorsal signals 11.8 Spinal cord motor neurons at different dorso-ventral positions project to different trunk and limb muscles 11.9 Antero-posterior pattern in the spinal cord is determined in response to secreted signals from the node and adjacent mesoderm Summary The formation and migration of neurons 11.10 Neurons in Drosophila arise from proneural clusters 11.11 The development of neurons in Drosophila involves asymmetric cell divisions and timed changes in gene expression 11.12 The production of vertebrate neurons involves lateral inhibition, as in Drosophila 11.13 Neurons are formed in the proliferative zone of the vertebrate neural tube and migrate outwards 11.14 Many cortical interneurons migrate tangentially Summary Axon navigation 11.15 The growth cone controls the path taken by a growing axon 11.16 Motor neuron axons in the chick limb are guided by ephrin–Eph interactions 11.17 Axons crossing the midline are both attracted and repelled 11.18 Neurons from the retina make ordered connections with visual centers in the brain Summary Synapse formation and refinement 11.19 Synapse formation involves reciprocal interactions 11.20 Many motor neurons die during normal development 11.21 Neuronal cell death and survival involve both intrinsic and extrinsic factors 11.22 The map from eye to brain is refined by neural activity Summary Summary to Chapter 11 12 Growth, post-embryonic development, and regeneration Growth 12.1 Tissues can grow by cell proliferation, cell enlargement, or accretion 12.2 Cell proliferation is controlled by regulating entry into the cell cycle 12.3 Cell division in early development can be controlled by an intrinsic developmental program 12.4 Extrinsic signals coordinate cell division, cell growth, and cell death in the developing Drosophila wing 12.5 Cancer can result from mutations in genes that control cell proliferation 12.6 The relative contributions of intrinsic and extrinsic factors in controlling size differ in different mammalian organs 12.7 Overall body size depends on the extent and the duration of growth 12.8 Hormones and growth factors coordinate the growth of different tissues and organs and contribute to determining overall body size 12.9 Elongation of the long bones illustrates how growth can be determined by a combination of an intrinsic growth program and extracellular factors 12.10 The amount of nourishment an embryo receives can have profound effects in later life Summary Molting and metamorphosis 12.11 Arthropods have to molt in order to grow 12.12 Insect body size is determined by the rate and duration of larval growth 12.13 Metamorphosis in amphibians is under hormonal control Summary Regeneration 12.14 Regeneration involves repatterning of existing tissues and/or growth of new tissues 12.15 Amphibian limb regeneration involves cell dedifferentiation and new growth 12.16 Limb regeneration in amphibians depends on the presence of nerves 12.17 The limb blastema gives rise to structures with positional values distal to the site of amputation 12.19 Mammals can regenerate the tips of the digits 12.20 Insect limbs intercalate positional values by both proximo-distal and circumferential growth 12.21 Heart regeneration in zebrafish involves the resumption of cell division by cardiomyocytes Summary Aging and senescence 12.22 Genes can alter the timing of senescence 12.23 Cell senescence blocks cell proliferation 12.24 Elimination of senescent cells in adult salamanders explains why regenerative ability does not diminish with age Summary Summary to Chapter 12 13 Plant development 13.1 The model plant Arabidopsis thaliana has a short life cycle and a small diploid genome Embryonic development 13.2 Plant embryos develop through several distinct stages 13.3 Gradients of the signal molecule auxin establish the embryonic apical–basal axis 13.4 Plant somatic cells can give rise to embryos and seedlings 13.5 Cell enlargement is a major process in plant growth and morphogenesis Summary Meristems 13.6 A meristem contains a small, central zone of self-renewing stem cells 13.7 The size of the stem cell area in the meristem is kept constant by a feedback loop to the organizing center 13.8 The fate of cells from different meristem layers can be changed by changing their position 13.9 A fate map for the embryonic shoot meristem can be deduced using clonal analysis 13.10 Meristem development is dependent on signals from other parts of the plant 13.11 Gene activity patterns the proximo-distal and adaxial–abaxial axes of leaves developing from the shoot meristem 13.12 The regular arrangement of leaves on a stem is generated by regulated auxin transport 13.13 The outgrowth of secondary shoots is under hormonal control 13.14 Root tissues are produced from Arabidopsis root apical meristems by a highly stereotyped pattern of cell divisions 13.15 Root hairs are specified by a combination of positional information and lateral inhibition Summary Flower development and control of flowering 13.16 Homeotic genes control organ identity in the flower 13.17 The Antirrhinum flower is patterned dorso-ventrally, as well as radially 13.18 The internal meristem layer can specify floral meristem patterning 13.19 The transition of a shoot meristem to a floral meristem is under environmental and genetic control 13.20 Vernalization reflects the epigenetic memory of winter 13.21 Most flowering plants are hermaphrodites, but some produce unisexual flowers Summary Summary to Chapter 13 14 Evolution and development The evolution of development 14.1 Multicellular organisms evolved from single-celled ancestors 14.2 Genomic evidence is throwing light on the evolution of animals 14.3 How gastrulation evolved is not known 14.4 More general characteristics of the body plan develop earlier than specializations 14.5 Embryonic structures have acquired new functions during evolution 14.6 Evolution of different types of eyes in different animal groups is an example of parallel evolution Summary The diversification of body plans 14.7 Hox gene complexes have evolved through gene duplication 14.9 Changes in Hox gene expression and their target genes contributed to the evolution of the vertebrate axial skeleton 14.10 The basic body plan of arthropods and vertebrates is similar, but the dorso-ventral axis is inverted Summary The evolutionary modification of specialized characters 14.11 Limbs evolved from fins 14.12 Limbs have evolved to fulfill different specialized functions 14.13 The evolution of limblessness in snakes is associated with changes in axial gene expression and mutations in a limb-specific enhancer 14.14 Butterfly wing markings have evolved by redeployment of genes previously used for other functions 14.15 Adaptive evolution within the same species provides a way of studying the developmental basis for evolutionary change Summary Changes in the timing of developmental processes 14.16 Changes in growth can modify the basic body plan 14.17 Evolution can be due to changes in the timing of developmental events 14.18 The evolution of life histories has implications for developm [Show More]
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