Sialidase Activity of Influenza A Virus in an Endocytic Pathway Enhances Viral Replication
Identifieur interne : 001603 ( Main/Merge ); précédent : 001602; suivant : 001604Sialidase Activity of Influenza A Virus in an Endocytic Pathway Enhances Viral Replication
Auteurs : Takashi Suzuki ; Tadanobu Takahashi ; Chao-Tan Guo ; Kazuya I.-P. Jwa Hidari ; Daisei Miyamoto ; Hideo Goto ; Yoshihiro Kawaoka [États-Unis] ; Yasuo SuzukiSource :
- Journal of Virology [ 0022-538X ] ; 2005.
Descripteurs français
- KwdFr :
- Acides sialiques (pharmacologie), Animaux, Antienzymes (pharmacologie), Chiens, Concentration en ions d'hydrogène, Endocytose, Grippe humaine (virologie), Guanidines, Humains, Lignée cellulaire, Mâle, Pyrannes, Réplication virale (), Réplication virale (génétique), Réplication virale (physiologie), Sialidase (antagonistes et inhibiteurs), Sialidase (génétique), Sialidase (métabolisme), Souris, Souris de lignée C57BL, Stabilité enzymatique, Transfection, Virus de la grippe A (), Virus de la grippe A (enzymologie), Virus de la grippe A (génétique), Virus de la grippe A (physiologie), Zanamivir.
- MESH :
- antagonistes et inhibiteurs : Sialidase.
- enzymologie : Virus de la grippe A.
- génétique : Réplication virale, Sialidase, Virus de la grippe A.
- métabolisme : Sialidase.
- pharmacologie : Acides sialiques, Antienzymes.
- physiologie : Réplication virale, Virus de la grippe A.
- virologie : Grippe humaine.
- Animaux, Chiens, Concentration en ions d'hydrogène, Endocytose, Guanidines, Humains, Lignée cellulaire, Mâle, Pyrannes, Réplication virale, Souris, Souris de lignée C57BL, Stabilité enzymatique, Transfection, Virus de la grippe A, Zanamivir.
English descriptors
- KwdEn :
- Animals, Cell Line, Dogs, Endocytosis, Enzyme Inhibitors (pharmacology), Enzyme Stability, Guanidines, Humans, Hydrogen-Ion Concentration, Influenza A virus (drug effects), Influenza A virus (enzymology), Influenza A virus (genetics), Influenza A virus (physiology), Influenza, Human (virology), Male, Mice, Mice, Inbred C57BL, Neuraminidase (antagonists & inhibitors), Neuraminidase (genetics), Neuraminidase (metabolism), Pyrans, Sialic Acids (pharmacology), Transfection, Virus Replication (drug effects), Virus Replication (genetics), Virus Replication (physiology), Zanamivir.
- MESH :
- chemical , antagonists & inhibitors : Neuraminidase.
- chemical , genetics : Neuraminidase.
- chemical , metabolism : Neuraminidase.
- chemical , pharmacology : Enzyme Inhibitors, Sialic Acids.
- drug effects : Influenza A virus, Virus Replication.
- enzymology : Influenza A virus.
- genetics : Influenza A virus, Virus Replication.
- physiology : Influenza A virus, Virus Replication.
- virology : Influenza, Human.
- Animals, Cell Line, Dogs, Endocytosis, Enzyme Stability, Guanidines, Humans, Hydrogen-Ion Concentration, Male, Mice, Mice, Inbred C57BL, Pyrans, Transfection, Zanamivir.
Abstract
N2 neuraminidase (NA) genes of the 1957 and 1968 pandemic influenza virus strains possessed avian-like low-pH stability of sialidase activity, unlike most epidemic strains. We generated four reverse-genetics viruses from a genetic background of A/WSN/33 (H1N1) that included parental N2 NAs of 1968 pandemic (H3N2) and epidemic (H2N2) strains or their counterpart N2 NAs in which the low-pH stability of the sialidase activity was changed by substitutions of one or two amino acid residues. We found that the transfectant viruses bearing low-pH-stable sialidase (WSN/Stable-NAs) showed 25- to 80-times-greater ability to replicate in Madin-Darby canine kidney (MDCK) cells than did the transfectant viruses bearing low-pH-unstable sialidase (WSN/Unstable-NAs). Enzymatic activities of WSN/Stable-NAs were detected in endosomes of MDCK cells after 90 min of virus internalization by in situ fluorescent detection with 5-bromo-4-chloro-indole-3-yl-α-
Url:
DOI: 10.1128/JVI.79.18.11705-11715.2005
PubMed: 16140748
PubMed Central: 1212606
Links toward previous steps (curation, corpus...)
- to stream Pmc, to step Corpus: 000381
- to stream Pmc, to step Curation: 000381
- to stream Pmc, to step Checkpoint: 000C25
- to stream PubMed, to step Corpus: 000318
- to stream PubMed, to step Curation: 000318
- to stream PubMed, to step Checkpoint: 000288
- to stream Ncbi, to step Merge: 000135
- to stream Ncbi, to step Curation: 000135
- to stream Ncbi, to step Checkpoint: 000135
Links to Exploration step
PMC:1212606Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Sialidase Activity of Influenza A Virus in an Endocytic Pathway Enhances Viral Replication</title>
<author><name sortKey="Suzuki, Takashi" sort="Suzuki, Takashi" uniqKey="Suzuki T" first="Takashi" last="Suzuki">Takashi Suzuki</name>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
</author>
<author><name sortKey="Takahashi, Tadanobu" sort="Takahashi, Tadanobu" uniqKey="Takahashi T" first="Tadanobu" last="Takahashi">Tadanobu Takahashi</name>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
</author>
<author><name sortKey="Guo, Chao Tan" sort="Guo, Chao Tan" uniqKey="Guo C" first="Chao-Tan" last="Guo">Chao-Tan Guo</name>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
</author>
<author><name sortKey="Hidari, Kazuya I P Jwa" sort="Hidari, Kazuya I P Jwa" uniqKey="Hidari K" first="Kazuya I.-P. Jwa" last="Hidari">Kazuya I.-P. Jwa Hidari</name>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
</author>
<author><name sortKey="Miyamoto, Daisei" sort="Miyamoto, Daisei" uniqKey="Miyamoto D" first="Daisei" last="Miyamoto">Daisei Miyamoto</name>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
</author>
<author><name sortKey="Goto, Hideo" sort="Goto, Hideo" uniqKey="Goto H" first="Hideo" last="Goto">Hideo Goto</name>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
</author>
<author><name sortKey="Kawaoka, Yoshihiro" sort="Kawaoka, Yoshihiro" uniqKey="Kawaoka Y" first="Yoshihiro" last="Kawaoka">Yoshihiro Kawaoka</name>
<affiliation><nlm:aff id="aff1"></nlm:aff>
<country>États-Unis</country>
<placeName><settlement type="city">Madison (Wisconsin)</settlement>
<region type="state">Wisconsin</region>
</placeName>
<orgName type="university" n="3">Université du Wisconsin à Madison</orgName>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
<country>États-Unis</country>
<placeName><settlement type="city">Madison (Wisconsin)</settlement>
<region type="state">Wisconsin</region>
</placeName>
<orgName type="university" n="3">Université du Wisconsin à Madison</orgName>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
<country>États-Unis</country>
<placeName><settlement type="city">Madison (Wisconsin)</settlement>
<region type="state">Wisconsin</region>
</placeName>
<orgName type="university" n="3">Université du Wisconsin à Madison</orgName>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
<country>États-Unis</country>
<placeName><settlement type="city">Madison (Wisconsin)</settlement>
<region type="state">Wisconsin</region>
</placeName>
<orgName type="university" n="3">Université du Wisconsin à Madison</orgName>
</affiliation>
</author>
<author><name sortKey="Suzuki, Yasuo" sort="Suzuki, Yasuo" uniqKey="Suzuki Y" first="Yasuo" last="Suzuki">Yasuo Suzuki</name>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PMC</idno>
<idno type="pmid">16140748</idno>
<idno type="pmc">1212606</idno>
<idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1212606</idno>
<idno type="RBID">PMC:1212606</idno>
<idno type="doi">10.1128/JVI.79.18.11705-11715.2005</idno>
<date when="2005">2005</date>
<idno type="wicri:Area/Pmc/Corpus">000381</idno>
<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000381</idno>
<idno type="wicri:Area/Pmc/Curation">000381</idno>
<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000381</idno>
<idno type="wicri:Area/Pmc/Checkpoint">000C25</idno>
<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000C25</idno>
<idno type="wicri:source">PubMed</idno>
<idno type="RBID">pubmed:16140748</idno>
<idno type="wicri:Area/PubMed/Corpus">000318</idno>
<idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">000318</idno>
<idno type="wicri:Area/PubMed/Curation">000318</idno>
<idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">000318</idno>
<idno type="wicri:Area/PubMed/Checkpoint">000288</idno>
<idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">000288</idno>
<idno type="wicri:Area/Ncbi/Merge">000135</idno>
<idno type="wicri:Area/Ncbi/Curation">000135</idno>
<idno type="wicri:Area/Ncbi/Checkpoint">000135</idno>
<idno type="wicri:doubleKey">0022-538X:2005:Suzuki T:sialidase:activity:of</idno>
<idno type="wicri:Area/Main/Merge">001603</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Sialidase Activity of Influenza A Virus in an Endocytic Pathway Enhances Viral Replication</title>
<author><name sortKey="Suzuki, Takashi" sort="Suzuki, Takashi" uniqKey="Suzuki T" first="Takashi" last="Suzuki">Takashi Suzuki</name>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
</author>
<author><name sortKey="Takahashi, Tadanobu" sort="Takahashi, Tadanobu" uniqKey="Takahashi T" first="Tadanobu" last="Takahashi">Tadanobu Takahashi</name>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
</author>
<author><name sortKey="Guo, Chao Tan" sort="Guo, Chao Tan" uniqKey="Guo C" first="Chao-Tan" last="Guo">Chao-Tan Guo</name>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
</author>
<author><name sortKey="Hidari, Kazuya I P Jwa" sort="Hidari, Kazuya I P Jwa" uniqKey="Hidari K" first="Kazuya I.-P. Jwa" last="Hidari">Kazuya I.-P. Jwa Hidari</name>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
</author>
<author><name sortKey="Miyamoto, Daisei" sort="Miyamoto, Daisei" uniqKey="Miyamoto D" first="Daisei" last="Miyamoto">Daisei Miyamoto</name>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
</author>
<author><name sortKey="Goto, Hideo" sort="Goto, Hideo" uniqKey="Goto H" first="Hideo" last="Goto">Hideo Goto</name>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
</author>
<author><name sortKey="Kawaoka, Yoshihiro" sort="Kawaoka, Yoshihiro" uniqKey="Kawaoka Y" first="Yoshihiro" last="Kawaoka">Yoshihiro Kawaoka</name>
<affiliation><nlm:aff id="aff1"></nlm:aff>
<country>États-Unis</country>
<placeName><settlement type="city">Madison (Wisconsin)</settlement>
<region type="state">Wisconsin</region>
</placeName>
<orgName type="university" n="3">Université du Wisconsin à Madison</orgName>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
<country>États-Unis</country>
<placeName><settlement type="city">Madison (Wisconsin)</settlement>
<region type="state">Wisconsin</region>
</placeName>
<orgName type="university" n="3">Université du Wisconsin à Madison</orgName>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
<country>États-Unis</country>
<placeName><settlement type="city">Madison (Wisconsin)</settlement>
<region type="state">Wisconsin</region>
</placeName>
<orgName type="university" n="3">Université du Wisconsin à Madison</orgName>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
<country>États-Unis</country>
<placeName><settlement type="city">Madison (Wisconsin)</settlement>
<region type="state">Wisconsin</region>
</placeName>
<orgName type="university" n="3">Université du Wisconsin à Madison</orgName>
</affiliation>
</author>
<author><name sortKey="Suzuki, Yasuo" sort="Suzuki, Yasuo" uniqKey="Suzuki Y" first="Yasuo" last="Suzuki">Yasuo Suzuki</name>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff1"></nlm:aff>
</affiliation>
</author>
</analytic>
<series><title level="j">Journal of Virology</title>
<idno type="ISSN">0022-538X</idno>
<idno type="eISSN">1098-5514</idno>
<imprint><date when="2005">2005</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term>
<term>Cell Line</term>
<term>Dogs</term>
<term>Endocytosis</term>
<term>Enzyme Inhibitors (pharmacology)</term>
<term>Enzyme Stability</term>
<term>Guanidines</term>
<term>Humans</term>
<term>Hydrogen-Ion Concentration</term>
<term>Influenza A virus (drug effects)</term>
<term>Influenza A virus (enzymology)</term>
<term>Influenza A virus (genetics)</term>
<term>Influenza A virus (physiology)</term>
<term>Influenza, Human (virology)</term>
<term>Male</term>
<term>Mice</term>
<term>Mice, Inbred C57BL</term>
<term>Neuraminidase (antagonists & inhibitors)</term>
<term>Neuraminidase (genetics)</term>
<term>Neuraminidase (metabolism)</term>
<term>Pyrans</term>
<term>Sialic Acids (pharmacology)</term>
<term>Transfection</term>
<term>Virus Replication (drug effects)</term>
<term>Virus Replication (genetics)</term>
<term>Virus Replication (physiology)</term>
<term>Zanamivir</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Acides sialiques (pharmacologie)</term>
<term>Animaux</term>
<term>Antienzymes (pharmacologie)</term>
<term>Chiens</term>
<term>Concentration en ions d'hydrogène</term>
<term>Endocytose</term>
<term>Grippe humaine (virologie)</term>
<term>Guanidines</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Mâle</term>
<term>Pyrannes</term>
<term>Réplication virale ()</term>
<term>Réplication virale (génétique)</term>
<term>Réplication virale (physiologie)</term>
<term>Sialidase (antagonistes et inhibiteurs)</term>
<term>Sialidase (génétique)</term>
<term>Sialidase (métabolisme)</term>
<term>Souris</term>
<term>Souris de lignée C57BL</term>
<term>Stabilité enzymatique</term>
<term>Transfection</term>
<term>Virus de la grippe A ()</term>
<term>Virus de la grippe A (enzymologie)</term>
<term>Virus de la grippe A (génétique)</term>
<term>Virus de la grippe A (physiologie)</term>
<term>Zanamivir</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="antagonists & inhibitors" xml:lang="en"><term>Neuraminidase</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Neuraminidase</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Neuraminidase</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Enzyme Inhibitors</term>
<term>Sialic Acids</term>
</keywords>
<keywords scheme="MESH" qualifier="antagonistes et inhibiteurs" xml:lang="fr"><term>Sialidase</term>
</keywords>
<keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>Influenza A virus</term>
<term>Virus Replication</term>
</keywords>
<keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Virus de la grippe A</term>
</keywords>
<keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Influenza A virus</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Influenza A virus</term>
<term>Virus Replication</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Réplication virale</term>
<term>Sialidase</term>
<term>Virus de la grippe A</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Sialidase</term>
</keywords>
<keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Acides sialiques</term>
<term>Antienzymes</term>
</keywords>
<keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Réplication virale</term>
<term>Virus de la grippe A</term>
</keywords>
<keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Influenza A virus</term>
<term>Virus Replication</term>
</keywords>
<keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Grippe humaine</term>
</keywords>
<keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Influenza, Human</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Cell Line</term>
<term>Dogs</term>
<term>Endocytosis</term>
<term>Enzyme Stability</term>
<term>Guanidines</term>
<term>Humans</term>
<term>Hydrogen-Ion Concentration</term>
<term>Male</term>
<term>Mice</term>
<term>Mice, Inbred C57BL</term>
<term>Pyrans</term>
<term>Transfection</term>
<term>Zanamivir</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Animaux</term>
<term>Chiens</term>
<term>Concentration en ions d'hydrogène</term>
<term>Endocytose</term>
<term>Guanidines</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Mâle</term>
<term>Pyrannes</term>
<term>Réplication virale</term>
<term>Souris</term>
<term>Souris de lignée C57BL</term>
<term>Stabilité enzymatique</term>
<term>Transfection</term>
<term>Virus de la grippe A</term>
<term>Zanamivir</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en"><p>N2 neuraminidase (NA) genes of the 1957 and 1968 pandemic influenza virus strains possessed avian-like low-pH stability of sialidase activity, unlike most epidemic strains. We generated four reverse-genetics viruses from a genetic background of A/WSN/33 (H1N1) that included parental N2 NAs of 1968 pandemic (H3N2) and epidemic (H2N2) strains or their counterpart N2 NAs in which the low-pH stability of the sialidase activity was changed by substitutions of one or two amino acid residues. We found that the transfectant viruses bearing low-pH-stable sialidase (WSN/Stable-NAs) showed 25- to 80-times-greater ability to replicate in Madin-Darby canine kidney (MDCK) cells than did the transfectant viruses bearing low-pH-unstable sialidase (WSN/Unstable-NAs). Enzymatic activities of WSN/Stable-NAs were detected in endosomes of MDCK cells after 90 min of virus internalization by in situ fluorescent detection with 5-bromo-4-chloro-indole-3-yl-α-<italic>N</italic>
-acetylneuraminic acid and Fast Red Violet LB. Inhibition of sialidase activity of WSN/Stable-NAs on the endocytic pathway by pretreatment with 4-guanidino-2,4-dideoxy-<italic>N</italic>
-acetylneuraminic acid (zanamivir) resulted in a significant decrease in progeny viruses. In contrast, the enzymatic activities of WSN/Unstable-NAs, the replication of which had no effect on pretreatment with zanamivir, were undetectable in cells under the same conditions. Hemadsorption assays of transfectant-virus-infected cells revealed that the low-pH stability of the sialidase had no effect on the process of removal of sialic acid from hemagglutinin in the Golgi regions. Moreover, high titers of viruses were recovered from the lungs of mice infected with WSN/Stable-NAs on day 3 after intranasal inoculation, but WSN/Unstable-NAs were cleared from the lungs of the mice. These results indicate that sialidase activity in late endosome/lysosome traffic enhances influenza A virus replication.</p>
</div>
</front>
</TEI>
</record>
Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/H2N2V1/Data/Main/Merge
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 001603 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Merge/biblio.hfd -nk 001603 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien |wiki= Sante |area= H2N2V1 |flux= Main |étape= Merge |type= RBID |clé= PMC:1212606 |texte= Sialidase Activity of Influenza A Virus in an Endocytic Pathway Enhances Viral Replication }}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Merge/RBID.i -Sk "pubmed:16140748" \ | HfdSelect -Kh $EXPLOR_AREA/Data/Main/Merge/biblio.hfd \ | NlmPubMed2Wicri -a H2N2V1
This area was generated with Dilib version V0.6.33. |