Peroxisomes in Dermatology.ppt

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Peroxisomes Peroxisome Proliferator-Activator
Receptors in Dermatology

• M.Y.ABDEL_MAWLA,MD
• Zagazig Faculty of Medicine,Zagzig,EGYPT

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Peroxisomes in Cells

• The trouble with simple things is that one must
• understand them very well !!!

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The Peroxisome

• Single membrane
• Roughly spherical
• 0.2 - 1.7?m
• Composition varies

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Peroxisomes

• Peroxisomes intracellular organelles with
  important roles defined in many metabolic
  processes.
• Peroxisomesin all mamalian cells,including
  kertatinocytes
• They derive their name from their ability to
  produce H2O2 through a group of oxidizing enzymes
  which use molecular oxygen to transform their
  substrates, releasing H2O2 and OH

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Peroxisomes

• The oxidative stress resulting from H2O2 is known
  to stimulate phospholipase D, associated with the
  production of phosphatidic acid and
  diacylglycerol.
• These in turn affect adenylyl cyclase and protein
  kinase C, respectively, which can modulate a wide
  array of target proteins including plasma
  membrane receptors, contractile proteins and
  regulatory enzymes.

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Peroxisomes

• Peroxisomes are similar physically to lysosomes,
  but they are different in two important ways
• First, they are believed to be formed by
  self-replication (or perhaps by budding off from
  the smooth endoplasmic reticulum) rather than
  from the Golgi apparatus.
• Second, they contain oxidases rather than
  hydrolases.

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Peroxisomes Biogenesis

• The development of peroxisomes involves a series
  of events including the recruitment of
  (phospho)lipids, synthesis, sorting and
  assembly/activation of matrix membrane proteins.
• The peroxisomal membrane mainly consists of
  phosphatidylcholine phosphatidylethanolamine
  and largely resembles that of the endoplasmic
  reticulum (ER)

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Peroxisome Biogenesis
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Peroxisomes Biogenesis

• Membrane proteins are synthesized on free
  polysomes in the cytosol.
• The model discriminates between membrane proteins
  essential for the biogenesis of organelles (PMBs)
  and those involved in the organellar function
  (PMFs, e.g. transporters).
• PMBs (e.g. Pex2p, Pex3p, Pex15p and Pex16p) may
  initially be transported to the ER and,
  eventually vesicle-mediated, be sorted to the
  growing peroxisome
• PMFs may be transported directly to the organelle

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Peroxisomes Functions

• Peroxisomes play only a minor role in cellular
  functions.
• Peroxisomes play an important role in regulating
  cellular proliferation and differentiation as
  well as in the modulation of inflammatory
  mediators.
• Peroxisomes have broad effects on the metabolism
  of lipids, hormones.
• Peroxisomes also affect cellular membranes and
  adipocyte formation, as well as insulin
  sensitivity.
• Peroxisomes play a role in aging and
  tumorigenesis through their effects on oxidative
  stress.

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Peroxisomes Functions

• Peroxisomes may be either catabolic or anabolic.
• Some functions are exclusively peroxisomal
• Others shared with other cellular compartments
  (microsomes endoplasmic reticulum, and
  mitochondria).

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Peroxisomes Functions in Cell signaling

• N nucleus, MIT mitochondria, MIC microsomes,
  Per peroxisome, PKO phosphatidyl choline,
• PC phosphosphatidyl choline, AA arachidonic
  acid, OX peroxisomal oxidases,
• LCFA long-chain fatty acids, MCFA
  medium-chain fatty acids, DG diacylglycerol, PA
  
• phosphatidic acid, CAT catalase, OFR
  oxygen free radicals.
• Peroxisome functions involved n cell signaling
  would potentiate differentiation, while reactive
  oxygen species would potentiate tumorigenesis and
  aging and catalase and other peroxisomal
  scavengers would moderate the effects of reactive
  oxygen species

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Peroxisomes Free Radicles
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Peroxisomal Enzymes
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Key peroxisomal enzymes
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How Peroxisomes to Function?

• Peroxisomes, contribute to key metabolic pathways
  of the cell function.
• These organelles are able to proliferate
  abundantly upon exposure to so-called peroxisome
  proliferators.
• Peroxisome proliferators may activate PPARs by
  binding directly to the receptors,and the
  activated PPARs may regulate the expression of
  genes involved in lipid metabolism and peroxisome
  proliferation
• Peroxisome-located enzymes are also regulated by
  many molecules including nutrients (fatty acids,
  steroids), hormones (T3, retinoids), PPARs and
  other nuclear signaling factors.

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How Peroxisomes to Function?

• Peroxisomal proliferator activator receptors
  (PPARs) and ligands for these receptors modulate
  different peroxisomal functions.

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PPARs Distribution

• PPARs are found mainly in tissues associated with
  high fatty acid metabolism.
• Thus are expressed mainly in liver,
• They are also found in kidney, muscle, heart,
  fat, B and T lymphocytes , vascular smooth
  muscle and keratinocytes

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Peroxisome Proliferator-Activator Receptors
(PPARs)

• Nuclear hormone receptor superfamily
• Multiple isoforms (?, ? (d), ?)
• Unique tissue distribution

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Mechanism of PPAR action
?
Co-repressors Co-activators
PPARRXR?
?
?
Modulation of gene transcription
Biological effect
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Mechanism of PPAR action
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PPARs FUNCTIONS

• PPARs play a significant role in many in skin
  homeostasis.
• On binding of ligand, PPARs undergo a
  conformational change enabling them to form a
  heterodimer with the 9-cis retinoic acid receptor
  (RXR).
• PPAR/RXR complex forms a transcription factor
  able to bind to DNA at PPREs, which are DNA
  sequences specifically capable of recognition
  and binding by the DNA-binding component of this
  transcription factor.

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PPAR FUNCTIONS

• A large number of PPAR target genes have been
  identified with wide-ranging effects on cell
  proliferation differentiation, inflammatory
  responses and angiogenesis, as well as lipid and
  glucose metabolism.

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• Peroxisome proliferator-activated receptors
  (PPARs) function as heterodimers with retinoid X
  receptors (RXRs) and are activated by specific
  ligands they then modulate DNA transcription by
  binding to defined nucleotide sequences
  (peroxisome proliferator response element, PPRE)
  in the promoter region of target genes. Several
  cofactors (coactivators or corepressors) mediate
  the ability of nuclear receptors to stimulate or
  repress the transcription process. (b) The
  N-terminus A/B domain contains a
  ligand-independent transcriptional activation
  domain (AF-1), which can be regulated by
  mitogen-activated protein kinase (MAPK)
  phosphorylation in a and ? isotypes. The C domain
  contains two zinc-finger-like motifs that
  specifically bind the PPRE in the regulatory
  region of PPAR-responsive genes. The D domain or
  hinge region allows conformational changes in the
  molecule. The E/F domain consists of the
  ligand-binding domain and the ligand-dependent
  transcriptional activation domain (AF-2). The
  ligand-binding pocket appears to be quite large
  in comparison with other nuclear receptors,
  allowing the PPARs to interact with a broad range
  of natural and synthetic ligands

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ACTIVATION OF PPARs

• Most nuclear hormone receptors exist in an
  inactive form complexed with heats hock proteins
• PPAR activation and regulation may occur either
  following interaction with specific ligands or by
  kinase-mediated phosphorylation, following a
  variety of extracellular signals.
• Following binding of ligand, the PPAR undergoes a
  conformational change and becomes able to
  interact through the E/F domain with RXR to form
  heterodimers.
• These dimers will then interact with the cognate
  recognition motif (PPREs) in the promoter region
  of relevant genes to activate transcription of
  target genes.

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ACTIVATION OF PPARs

• PPARs are held in inactive complex
  associated with heat shock proteins (HsP).
  Following binding with ligand or other activation
  signals .
• HsP is replaced with retinoic acid receptor
  (RXR) forming active transcription factor.
• The active PPAR/RXR heterodimer binds to PPAR
  response element AGGTCA X AGGTCA to initiate
  transcription of relevant genes.

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PPAR? activation.

• Upon ligand binding, the nuclear receptor PPAR?
  associates with nuclear receptor RXR as well as
  with coactivators and corepressors which are
  present in a cell type and state specific
  pattern. This complex binds to PPAR response
  elements to enhance or repress gene transcription.

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Biological roles of PPARa
PPARa mediates the induction of multiple enzymes
required to mobilize and transport fatty acids
from adipose stores to liver for catabolism.
Basis for therapeutic use in humans to lower
serum lipids.
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Biological roles of PPARb/d
Ligand activation of PPARb/d leads to terminal
differentiation of keratinocytes as shown by four
independent laboratories. Activation of PPARb/d
in skeletal muscle leads to increased catabolism
of fatty acids and improved insulin sensitivity.
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Biological roles of PPARg
The role of PPARg in carcinogenesis is also
controversial. There is evidence that activation
of PPARg can either potentiate or attenuate
cancer, but current consensus favors
attenuation.
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Orchestration of Immune Responses
CELLS Lymphocytes Monocytes/Macs Neutrophils Eosin
ophils Basophils Dendritic cells
MOLECULES Complement Lysozyme Inflammatory
mediators Chemokines Cytokines
TISSUES Thymus Spleen Lymph nodes Blood
Innate immunity Adaptive Immunity
PPARs are found in a number of immune cell types
and there is evidence that they could modulate a
number of different immune responses
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Role of PPARa in Immune Function

• Expressed in monocytes/macrophages, increased
  after treatment with phorbol ester
• PPARa ligands induce apoptosis in activated
  macrophages
• PPARa ligands decrease secretion of MMPs in
  LPS-treated monocytes
• PPARa ligands decrease NOS activity in
  macrophages
• BUT
• Natural PPARa ligands increase NOS activity in
  macrophages

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Role of PPARa in Immune Function

• Inflammatory response induced by LTB4 is enhanced
  in PPARa-null mice
• PPARa ligands can inhibit inflammatory cytokine
  production
• BUT
• PPARa ligands cause increase in serum TNFa after
  LPS

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Role of PPARa in Immune Function

• Reports suggest that PPARa ligands are
  anti-inflammatory but there are also some reports
  suggesting that PPARa ligands are pro-inflammatory

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Peroxisomes Inflammation

• PPAR-a activation transcription factor for
  peroxisomal metabolism for arachidonic-derived
  proinflammatory eicosanoids.
• ligands for this activation are the w-3 fatty
  acids, which may explain their antiinflammatory
  effects.

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Peroxisomes Inflammation

• PPARa repress NFkB transcription, signal
  transducer and activator of transcriptions
  (STATs) and AP-1 (Jun/ Fos) transcription and the
  downstream signaling mechanism.
• These effects result in the decreased production
  of inflammatory cytokines, protease production,
  and some mechanisms for proliferation and
  apoptotic signaling.

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Peroxisomes Inflammation

• Activation of PPARs also modulates endothelial
  cell adhesion molecules, and endothelial cells
  express both PPARa and PPAR gamma

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Peroxisomes INFLAMMATION

• The peroxisome proliferator-activated receptors
  (PPARs) a, ß/d, and ? are ligand-activated
  transcription factors belonging to the nuclear
  receptor superfamily.
• In addition to their regulatory role on lipid and
  glucose metabolism, they exert anti-inflammatory
  properties.
• In skin both PPAR-a and PPAR-ß/d regulate
  keratinocyte proliferation/differentiation and
  contribute to wound healing.
• The 3 PPAR isoforms are expressed by several cell
  types recruited into the dermis during
  inflammation.

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Effects on angiogenesis

• Vascular endothelial cell growth factor(VEGF) is
  a potent endothelial cell-specific mitogen.
• PPAR agonists have diverse effects on the
  expression of VEGF .
• PPAR gamma ligands increase the generation of
  VEGF , which might be expected to have
  pro-angiogenic effects.

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Peroxisomes and Sterol Metabolism

• The levels of 17b-hydroxy forms of sex steroids
  are regulated by the 17b-hydroxysteroid
  dehydrogenase (17b-
• HSD) family of proteins.
• The type IV enzyme, found primarily in
  peroxisomes, possesses a number of unique features

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Peroxisomes and Sterol Metabolism

• 17b-HSD IV is involved in degradation of
  branched-chain fatty acids and the side chain of
  cholesterol.
• Its expression is stimulated by PPARa ligands

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Peroxisomes in Epidermal Differentiation
Proliferation

• A well documented effects of PPARs on gene
  transcription associated with lipid metabolism
  energy homeostasis.
• Roles in epidermal maturation, repair and
  angiogenesis are highlighted.

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Peroxisomes in Epidermal Differentiation
Proliferation

• PPARa has a role in barrier development.
• Activators of PPARa, such as clofibrate, oleic
  acid and linoleic acid, accelerate the barrier
  development. as evidenced by
• decreased transepidermal water loss (TEWL)
• increased epidermal stratification
• the appearance of mature lamellae in the
  extracellular spaces of a multilayered stratum
  corneum.
• the induction of b-galactosidase and steroid
  sulphatase,
• accelerated the expression of profilaggrin, and
  its processing to filaggrin, and the expression
  of loricrin, a

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Peroxisomes Atopic Dermatitis

• Changes in immunoglobulin-associated
  transcription in atopic dermatitis may favour
• IgE over secretory immunoglobulin (multimeric
  IgM and IgA) expression in AD skin.
• Decreased PPAR activity appears common
• to both AD and psoriasis, and reduced
  cutaneous IFNa2 transcription also appears
  characteristic of AD

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Peroxisomes in Epidermal Differentiation
Proliferation

• PPARa ligands to promote epidermal
  differentiation,to restore epidermal homeostasis
  in hyperproliferative
• mouse epidermis and regulate apoptosis.
• PPARb / gammainduced expression of involucrin
  and transglutaminase( markers of keratinocyte
  differentiation)

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Wound healing

• PPARa and PPARb / gammaexpression is upregulated
  in the keratinocytes at the wound edge of the
  damaged skin.
• PPARa is re-expressed transiently in this area
  during the early inflammatory phase of the
  healing.
• Delays in wound healing parallel the pattern of
  PPAR expression of the respective PPAR isotypes
• PPARb upregulation linked to
• proinflammatory cytokines, such as
• interferon, tumour necrosis factor (TNF)-a.

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Wound healing

• PPARb a key mediator ofepidermal effects in
  wound healing by converting the extracellular
  inflammatory signal into an organized pattern of
  gene expression leading to survival, migration
  and differentiation of keratinocytes.

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Sebocyte glands

• Activation of PPAR gamma a by their respective
  specific ligands, stimulates lipid
• droplet accumulation in sebocytes.
• Because increased sebum production is an
  important element in the pathogenesis of acne
  vulgaris, development PPAR antagonists
  interfering selectively with sebum formation may
  have implications for the treatment of acne.

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Psoriasis

• The hallmarks of psoriasis are abnormal
  differentiation hyperproliferation of
  keratinocytes with inflammatory cell
  infiltration. These cellular changes are likely
  to find their explanation in activated T
  lymphocytes infiltrating the skin.

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Psoriasis

• PPARs a critical regulator of cutaneous
  homeostasis in psoriasis.
• In view of their prodifferentiating,
  antiproliferative immunomodulating effects,
  PPAR ligands may be interesting compounds for
  the treatment of epidermal disorders showing
  inflammation,hyperproliferation
• aberrant differentiation, such as psoriasis.

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Psoriasis

• PPAR gamma and some of its ligands, including two
  lipoxygenase metabolites, are induced by the T
  helper (TH)2 cytokine IL-4 in macrophages.
• PPAR gamma activation may also be an integral
  part of the effecter mechanism for downregulation
  TH1 dominate inflammatory reaction such as
  psoriasis and rheumatoid arthritis

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Psoriasis

• The expression of both PPARa gamma is decreased
  in epidermis , whereas the exact opposite happens
  with PPAR dela.
• Treatment by troglitazone, a specific PPAR gamma
  activator, inhibited the proliferation of both
  normal and psoriatic human keratinocytes

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Psoriasis

• There may be an association between
• psoriasis and the genes encoding PPARa or PPAR
  gamma

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Effects on angiogenesis

• Vascular endothelial cell growth factor(VEGF) is
  a potent endothelial cell-specific mitogen.
• PPAR agonists have diverse effects on the
  expression of VEGF .
• PPAR gamma ligands increase the generation of
  VEGF , which might be expected to have
  pro-angiogenic effects.

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Human immunodeficiencyvirus-1-protease inhibitor
associatedlipodystrophy

• HIV-1-protease inhibitor-associated lipodystrophy
  the result of impaired cellular retinoic acid
  binding
• protein type I (CRABP-1)-mediated 9-cis
  retinoic stimulation of PPARcRXR.
• Altered differentiation status of peripheral
  adipocytes in HIV-1-infected patients with
  protease inhibitor lipodystrophy is associated
  with greatly reduced sterol-regulatory
  element-binding protein 1c (SREBP1c) mRNA
  expression
• Reduced SREBP1 expression consequently alters the
  PPAR gamma activity, which may lead to
  lipodystrophy and to metabolic alterations.

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Peroxisome defects

• There are now many inherited disorders known to
  relate to peroxisome defects, frequently with
  significant cutaneous manifestations such as
• ichthyosis
• Recurrent ulceration,
• alopecia,
• follicular atrophoderma
• And photosensitivity,
• Thus ,it is suggested that modification of
  their
• activity may be of therapeutic benefit in
  the field of dermatology

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Summary

•   Peroxisome proliferator-activated receptors
  (PPARs) are ligand-activated transcription
  factors that regulate the expression of target
  genes involved in many cellular functions
  including cell proliferation, differentiation and
  immune/inflammation response.
• The PPAR subfamily consists of three isotypes
  PPARa, PPARß/d and PPAR?, which have all been
  identified in keratinocytes.
•  

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Summary

• PPARß/d is the predominant subtype in human
  keratinocytes, whereas PPARa and PPAR? are
  expressed at much lower levels and increase
  significantly upon keratinocyte differentiation.
• PPARß/d is significantly upregulated upon
  various conditions that result in keratinocyte
  proliferation, and during skin wound healing.

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Summary

• PPARs appear to play an important role in skin
  barrier permeability, inhibiting epidermal cell
  growth, promoting epidermal terminal
  differentiation and regulating skin inflammatory
  response by diverse mechanisms.
• These proprieties are pointing in the direction
  of PPARs being key regulators of skin conditions
  characterized by hyperproliferation, inflammatory
  infiltrates and aberrant differentiation such as
  psoriasis,
• They may also have clinical implications in
  other inflammatory skin disease (e.g. atopic
  dermatitis), proliferative skin disease, wound
  healing, acne and protease inhibitor associated
  lipodystrophia.

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A Message Home

• Peroxisomes are small cellular organelles that
  were almost ignored for years because they were
  believed to play only a minor role in cellular
  functions.
• Peroxisomes play an important role in regulating
  cellular proliferation and differentiation as
  well as in the modulation of inflammatory
  mediators.
• Peroxisomes have broad effects on the metabolism
  of lipids, hormones,.
• Through their effects on lipid metabolism,
  peroxisomes also affect cellular membranes and
  adipocyte formation, as well as insulin
  sensitivity
• Peroxisomes play a role in aging and
  tumorigenesis through their effects on oxidative
  stress.

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