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 https://mednexus.org/doi/10.1097/JBR.0000000000000039

Zinc-finger protein A20 protects hair cells from damage made by high-power microwave

Published Online: 17 September 2019
 
Inner ear hair cells are important for maintaining hearing. Irreversible damage to hair cells is an important cause of sensorineural deafness. Electromagnetic radiation, especially high-power microwave, is an important threat to human health in modern society and war. However, it is not clear whether high-power microwave has an effect on cochlea hair cells. This study aimed to assess the effects of high-power microwave on cochlear hair cells in guinea pigs, and investigate the potential protection of these cells against high-power microwave-induced damage by recombinant adenovirus A20. Based on experimental results, a 65 W/cm2 irradiation density applied to guinea pigs in this study to establish a high-power microwave inner ear injury model. In addition, pAdEeay-1/A20 was injected via a round window into experimental guinea pig cochlea, whereas artificial perilymph was injected into the control group. Auditory function was assessed by testing the auditory brainstem response threshold, and damage to cochlear hair cells was investigated by cell counting and scanning electron microscopy observations of the basilar membrane. Inner ear injury was observed 6 hours after 65 W/cm2 of irradiation and the auditory brainstem response threshold was significantly higher in the irradiation group (P < 0.05) compared with other groups. Propidium iodide staining and scanning electron microscopy results indicated that significant morphological changes occurred after radiation, especially to inner hair cells, which exhibited remarkable damage and the presence of several unknown spherical substances. Auditory brainstem response threshold was decreased in the pAdEeay-1/A20 group compared with the artificial perilymph group; moreover, damage to hair cells was milder in the pAdEeay-1/A20 group compared with the control group (P < 0.01). Thus, high-power microwave can cause damage to cochlear hair cells, as well as hearing loss with prolonged exposure and/or high dosage. In this regard, 65 W/cm2 of irradiation for 6 hours is a reliable target dose for observation of damage. The zinc finger protein A20 can protect cochlear hair cells from high-power microwave-induced damage and prevent further hearing loss. This study was approved by the Laboratory Animal Welfare and Ethics Committee of the Third Military Medical University, China on April 18, 2017.
Introduction
With the advance of industrial applications of radiation in modern society, humans are increasingly subjected to electromagnetic radiation.[1] In modern warfare, electromagnetic radiation, especially high-power microwave (HPM), poses a growing threat to human health. However, whether and how electromagnetic radiation can affect cochlear hair cells remains unclear. HPM refers to electromagnetic waves with peak power exceeding 100 MW and frequencies ranging from 1 to 300 GHz, thus spanning centimeter and millimeter bands. The biological effects of HPM represent a new, hot research topic that has accompanied the development of HPM weapons. Indeed, the advent of HPM has greatly attracted attention to its biological effects, as resulting injuries are not only caused by direct exposure to microwave weaponry, but also by frequent low-dose HPM exposure and accidental high-dose HPM leaks occurring during microwave weapon production  and training. Importantly, HPM contains some unique effects that are not readily observed with ordinary biological waves. To date, several studies have focused on the biological effects of microwaves on vital organs; however, no previous research examined the effects of HPM on the auditory system. Hence, it is essential to study the mechanisms by which HPM causes damage to cochlear hair cells. Abstract
HPM-induced damage to organisms has attracted significant attention form scholars. Indeed, previous studies have shown that HPM has harmful effects on several human organs and systems.[2,3] In particular, the nervous,[4] cardiovascular,[5] immune, and reproductive[6,7] systems are target organs that may be negatively affected by HPM, with adverse consequences such as nerve weakness, memory loss, myocardial injury, hematopoietic dysfunction, and declined sperm motility. The influence of microwave on auditory morphology has been gradually studied. Initially, Yu et al[8,9] found that a high-power electromagnetic pulse could lead to increased auditory brainstem response (ABR) thresholds in mice. Subsequently, Feng et al[10] reported that prolonged exposure to microwaves could lead to dysfunction of outer hair cells. Later, Seckin et al[11] demonstrated that radiofrequency radiation caused damage to cell structures of the cochlea during rat development. However, studies examining pathological changes elicited by HPM in cochlear hair cells and potential protective effects are still lacking. Hence, in this study, we established an HPM-induced hair cell injury model to observe the effects of HPM on guinea pig cochlear hair cells.
 
The zinc finger protein A20 was initially discovered in 1990 in human endothelial cells as a response product to tumor necrosis factor.[12] Through gene sequencing, it was found that its readable frame encoded a new zinc finger protein, named zinc finger protein A20 or just A20 for brevity. It has been revealed that A20 can inhibit nuclear factor-kappa B (NF-κB) and caspase-3 expression to protect cells form damage,[13,14] such as apoptosis of endothelial cells[15] and nerve cells.[16] Auditory hair cells are the primary sensory receptors of the mammalian cochlea, irreversible damage to hair cells is an important cause of sensorineural deafness. Previous research has shown that hair cell damage is mainly caused by caspase-3 activation, release of reactive oxygen species, and NF-κB activation.[17,18] This is consistent with the anti-damage mechanism of A20, so we chose the zinc finger protein A20 for this initial examination of a protective effect in HPM-induced hair cell injury model. We constructed a recombinant adenovirus pAdEeay-1/A20 containing the zinc finger protein A20 gene to investigate potential protection of hair cells from HPM-induced damage. Adenovirus is an effective vector for gene therapy of inner ear, and does not cause damage to inner ear function.[19] With regard to the importance of hearing both during peacetime and wartime, it is of great theoretical value and practical significance to study the effect of electromagnetic radiation on hearing function.

( Kommentti tässä välissä)

: Genecaards : HAKU  "A20"    VASTAUIS:    TNFAIP3             

A20,   Cr. 6q23.3 , Gene:, Zincfinger protein A20  
Recommended name: Tumor necrosis factor alpha-induced protein 3

KOMMENTTEJANI:  (Tuumorinekroosifaktori alfan  aiheuttama proteiini 3. Proteiinin  luonnetta ja ryhmää kuvaavia nimiä: OTUD7C, epätavallinen  ubikitinaasi-deubikitinaasi proteiini,  Omaa C-terminaalissa  7 sinkkisormea josta nimi: Zinc finger protein A20. 

  • GeneCards Symbol: TNFAIP3 2
  • TNF Alpha Induced Protein 3 2 3 5
  • OTUD7C 2 3 4 5
  • A20 2 3 5
  • Tumor Necrosis Factor Alpha-Induced Protein 3 3 4
  • OTU Domain-Containing Protein 7C 3 4
  • Putative DNA-Binding Protein A20 3 4
  • Zinc Finger Protein A20 3 4
  • Tumor Necrosis Factor, Alpha-Induced Protein 3
  •  Tumor Necrosis Factor, Alpha Induced Protein 3 3

    NCBI Gene Summary for TNFAIP3 Gene ( KOMMENTTINI geenistä::  Tämä geeni ilmentää proteiinia, jota indusoituu eli aiheutuu nopeasti  tuumorinekroositekijästä TNF.  Tämä proteiini  sinkkisormiproteiineja ja ubikitiinia editoiva entsyymi. Sen on havaittu estävän NFkappaB-aktivaatiota ja myös  TNF-välitteistä  apoptoosia eli solun ohjelmoitua kuolemaa. Tällä proteiinilla on sekä ubikitinaasin että deubikitinaasin aktiivisuudet ja se osallistuu sytokiinivälitteiseen immuniteettiin ja tulehdusvasteisiin. Sitä samaa proteiinia  kooda useahko  transkriptivariantti. (Proteiini on konstitutiivinen).

    • This gene was identified as a gene whose expression is rapidly induced by the tumor necrosis factor (TNF). The protein encoded by this gene is a zinc finger protein and ubiqitin-editing enzyme, and has been shown to inhibit NF-kappa B activation as well as TNF-mediated apoptosis. The encoded protein, which has both ubiquitin ligase and deubiquitinase activities, is involved in the cytokine-mediated immune and inflammatory responses. Several transcript variants encoding the same protein have been found for this gene. [provided by RefSeq, Jul 2012]

    GeneCards Summary for TNFAIP3 Gene  (Yhteenvetoa geenistä TNFAIP3-geeni on  proteiinia koodaava ja tuottaa TNFalfan indusoimaa proteiinia numero 3. Tunnetaan tauteja, jotka liittyvät tähän eeniin kuten periytyvä Behcetin kaltaisen  taudin tyyppi 1 kaltainen autoinflammatorinen oireyhtymä... Tähän geeniin liittyy  ligaasiaktiivisuus ja cysteiiityyppinen dewubikitinaasiaktiivisuus.

    TNFAIP3 (TNF Alpha Induced Protein 3) is a Protein Coding gene. Diseases associated with TNFAIP3 include Autoinflammatory Syndrome, Familial, Behcet-Like 1 and Familial Behcet-Like Autoinflammatory Syndrome. Among its related pathways are TNF signaling and Metabolism of proteins. Gene Ontology (GO) annotations related to this gene include ligase activity and cysteine-type deubiquitinase activity. An important paralog of this gene is OTUD7B.

    UniProtKB/Swiss-Prot Summary for TNFAIP3 Gene 
    (Yhteenvetoa proteiinista ja sen  funktiokartasta),  Tämä ubikitiiniä editoiva entsyymi omaa  sekä ubikitiiniligaasin   että deubikitinaasin aktiivisuuden. Se osallistuu  immuuni- ja tulehdusvasteisiin, joita signaloivat erilaiset sytokiinit kuten TNFalfa ja IL_1 beeta tai  TOLLin kaltaisten reseptoreiden  kautta  signaloivat  patogeenit ja se   pystyy  lopettamaan, saattamaan terminaatioon, . NF-kB aktiivisuuden.   

    Ubiquitin-editing enzyme that contains both ubiquitin ligase and deubiquitinase activities. Involved in immune and inflammatory responses signaled by cytokines, such as TNF-alpha and IL-1 beta, or pathogens via Toll-like receptors (TLRs) through terminating NF-kappa-B activity. 

     Tässä artikkelissa annetaan nimeltä tärkeitä ubikitiinia editoivien kompleksin välttämättömien jäsenten nimiä: kuten RNF11 ( RING finger proteiini11) , ITCH 

    Essential component of a ubiquitin-editing protein complex, comprising also RNF11, ITCH and TAX1BP1, that ensures the transient nature of inflammatory signaling pathways. In cooperation with TAX1BP1 promotes disassembly of E2-E3 ubiquitin protein ligase complexes in IL-1R and TNFR-1 pathways; affected are at least E3 ligases TRAF6, TRAF2 and BIRC2, and E2 ubiquitin-conjugating enzymes UBE2N and UBE2D3. In cooperation with TAX1BP1 promotes ubiquitination of UBE2N and proteasomal degradation of UBE2N and UBE2D3. Upon TNF stimulation, deubiquitinates 'Lys-63'-polyubiquitin chains on RIPK1 and catalyzes the formation of 'Lys-48'-polyubiquitin chains. This leads to RIPK1 proteasomal degradation and consequently termination of the TNF- or LPS-mediated activation of NF-kappa-B. Deubiquitinates TRAF6 probably acting on 'Lys-63'-linked polyubiquitin. Upon T-cell receptor (TCR)-mediated T-cell activation, deubiquitinates 'Lys-63'-polyubiquitin chains on MALT1 thereby mediating disassociation of the CBM (CARD11:BCL10:MALT1) and IKK complexes and preventing sustained IKK activation. Deubiquitinates NEMO/IKBKG; the function is facilitated by TNIP1 and leads to inhibition of NF-kappa-B activation. Upon stimulation by bacterial peptidoglycans, probably deubiquitinates RIPK2. Can also inhibit I-kappa-B-kinase (IKK) through a non-catalytic mechanism which involves polyubiquitin; polyubiquitin promotes association with IKBKG and prevents IKK MAP3K7-mediated phosphorylation. Targets TRAF2 for lysosomal degradation. In vitro able to deubiquitinate 'Lys-11'-, 'Lys-48'- and 'Lys-63' polyubiquitin chains. Inhibitor of programmed cell death. Has a role in the function of the lymphoid system. Required for LPS-induced production of pro-inflammatory cytokines and IFN beta in LPS-tolerized macrophages. ( TNAP3_HUMAN,P21580

    Protein attributes for TNFAIP3 Gene

    Size:
    790 amino acids
    Molecular mass:
    89614 Da
    Protein existence level:
    PE1
    Quaternary structure:


    Homodimer.
    Interacts with TNIP1, TAX1BP1 and TRAF2.
    Interacts with RNF11, ITCH and TAX1BP1 only after TNF stimulation; these interaction are transient and they are lost after 1 hour of stimulation with TNF (By similarity).
    Interacts with YWHAZ and YWHAH.
    Interacts with IKBKG; the interaction is induced by TNF stimulation and by polyubiquitin.
    Interacts with RIPK1.
    Interacts with UBE2N; the interaction requires TAX1BP1.
    Interacts with TRAF6; the interaction is inhibited by HTLV-1 protein Tax.

    Gene Families for TNFAIP3 Gene

    HGNC:
    The Human Protein Atlas (HPA):
    • Cancer-related genes
    • Disease related genes
    • Enzymes
    • Human disease related genes
    • Plasma proteins
    • Potential drug targets
    • Predicted intracellular proteins

    Protein Domains for TNFAIP3 Gene

    • InterPro:
    Blocks:
    • OTU-like cysteine protease
    • Zn-finger, A20-like

    (  Taulukossa kaikki OTU geenit, jotka ovat  deubikitinaaseja ("DUB")  mutta tämä  A20 omaa sekä UB ett DUB funktiota. A20 on " unclassified E3",  " Both Ligase AND DUB" ,  OTUD7C.
     (https://www.genenames.org/data/genegroup/#!/group/669 ). (Minulla on kaikki OTUS geenit vihkossa DUB, deubikitinaasit,  Ovarian tumor proteases  1 seulontaa. ).
    (1)  
    Tässä en ole asettanut muistiin minkälainen sinkkisormijärjestymä tähän  muodostuu.  Kompleksisssa näkyy tekstin mukaan  myös Ringfinger  olevan RNF11 (Lisätieto  tästä RING domeeni sinkkifinger proteiinista. https://www.genecards.org/cgi-bin/carddisp.pl?gene=RNF11&keywords=CGI-123
     
    RNF11  on  Essential component of a ubiquitin-editing protein complex, comprising also TNFAIP3 (Alias A20), ITCH (HECT ryhmän E3 ub. ligaasi)  and TAX1BP1, that ensures the transient nature of inflammatory signaling pathways. (Kommentti: Tässä katsoin tarkemmin  molekyyliä Tax1bp1 JA LÖYSIN SIITÄ DOMAANIN JOSSA ON SINKKISORMI.FUNKTIO: (ubz, UBIKITIINIÅ SITOVA SINKKISORMI, calcoco½). 
     
    JATKUU.. 
     
    Promotes the association of TNFAIP3 to RIPK1 after TNF stimulation. TNFAIP3 deubiquitinates 'Lys-63' polyubiquitin chains on RIPK1 and catalyzes the formation of 'Lys-48'-polyubiquitin chains. This leads to RIPK1 proteasomal degradation and consequently termination of the TNF- or LPS-mediated activation of NF-kappa-B. Recruits STAMBP to the E3 ubiquitin-ligase SMURF2 for ubiquitination, leading to its degradation by the 26S proteasome. ( RNF11_HUMAN,Q9Y3C5 )  
     Protein Domains for RNF11 Gene  
     
     (2) 
    (Lisätieto ITCH GEENISTÄ JA PROTEIINISTA.   Se tekee runsaasti interaktioita ja iso liuta  viruksia mainitaan sen interaktioitten yhteydessä. ITCH mainitaan essentielliksi  siinä  ubikitiinia editoivassa  kompleksissa, jossa toimii A20.
     ITCH kuuluu ubikitiinia editoivien entsyymien HECT ryhmään ja siinä on 4 WW tunnuksena. .  Komleksien täysi  monipuolinen jäsenmäärä vastaa tapahtumien  hetkellisyyksistä ja ohimenevyyksistä, "transient nature", mikä on plastista  biologista elämää kehossa. Esim kuten  hyvin toimiva yhteiskunta, jossa roskat heti poistetaan näkyvistä ja hyödynnetään, jolloin on joustavuutta ja valmiutta ja viihtyvyyttä).  Kun on tullut  merkki tulehduksesta TNF,  myös  tämä ItCH (
     
    Recommended name: E3 ubiquitin-protein ligase Itchy homolog
     #Involved in the control of inflammatory signaling pathways (PubMed:19131965). Essential component of a ubiquitin-editing protein complex, comprising also TNFAIP3 (A20), TAX1BP1 and RNF11, that ensures the transient nature of inflammatory signaling pathways (PubMed:19131965). Promotes the association of the complex after TNF stimulation (PubMed:19131965). Once the complex is formed, TNFAIP3 deubiquitinates 'Lys-63' polyubiquitin chains on RIPK1 and catalyzes the formation of 'Lys-48'-polyubiquitin chains (PubMed:19131965). This leads to RIPK1 proteasomal degradation and consequently termination of the TNF- or LPS-mediated activation of NFKB1" (PubMed:19131965).

    Gene Families for ITCH Gene

    HGNC:
    The Human Protein Atlas (HPA):
    • Disease related genes
    • Enzymes
    • Human disease related genes
    • Metabolic proteins
    • Potential drug targets
    • Predicted intracellular proteins

    Protein Domains for ITCH Gene

    InterPro:
    Blocks:
    • C2 domain
    • WW domain signature
    • HECT domain (Ubiquitin-protein ligase)
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