107-02-8HGINCPLSRVDWNT-UHFFFAOYSA-NHGINCPLSRVDWNT-UHFFFAOYSA-N
Acrolein2-Propenal
2-Propen-1-al
2-Propen-1-one
acrilaldehido
Acroleina
Acrylaldehyd
Acrylaldehyde
Acrylic aldehyde
Allyl aldehyde
Aqualin
Magnacide B
Magnacide H
NSC 8819
Prop-2-en-1-al
Propenal
UN 1092
DTXSID502002310028-15-6CBENFWSGALASAD-UHFFFAOYSA-NCBENFWSGALASAD-UHFFFAOYSA-N
OzoneAtmospheric ozone
Healozone
Oxygen, mol.
Ozone(16O16O16O)
Triatomic oxygen
DTXSID0021098NOCASCigarette smokeCS
DTXSID503503810102-44-0JCXJVPUVTGWSNB-UHFFFAOYSA-NJCXJVPUVTGWSNB-UHFFFAOYSA-N
Nitrogen dioxideBioxido de Nitrogeno
dioxido de nitrogeno
Dioxyde d'azote
Nitrite radical
Nitrito
NITROGEN PEROXIDE
Stickstoffdioxid
UN 1067
DTXSID7020974NOCASDiesel engine exhaustDiesel Exhaust
DE
DTXSID102404375-07-0IKHGUXGNUITLKF-UHFFFAOYSA-NIKHGUXGNUITLKF-UHFFFAOYSA-N
AcetaldehydeEthanal
acetaldehido
Acetaldehyd
Acetic aldehyde
Ethyl aldehyde
NSC 7594
UN 1089
DTXSID503922454-11-5SNICXCGAKADSCV-JTQLQIEISA-NSNICXCGAKADSCV-JTQLQIEISA-N
NicotineNicotine alkaloid
Pyridine, 3-[(2S)-1-methyl-2-pyrrolidinyl]-
(-)-(S)-Nicotine
(-)-3-(1-Methyl-2-pyrrolidyl)pyridine
(-)-Nicotine
(-)-β-Pyridyl-α-N-methylpyrrolidine
(S)-(-)-Nicotine
(S)-3-(1-Methyl-2-pyrrolidinyl)pyridine
(S)-Nicotine
3-[(2S)-1-Methyl-2-pyrrolidinyl]pyridine
Flux Maag
Habitrol
l-Nicotine
Nicabate
Nicoderm
Nicolan
Niconil
Nicopatch
Nicorette
Nicotell TTS
Nicotin
nicotina
NICOTINE,
Nicotinell
Nicotrol
NSC 5065
Pyridine, 3-(1-methyl-2-pyrrolidinyl)-, (S)-
Tabazur
UN 1654
XL All Insecticide
DTXSID10209307446-09-5RAHZWNYVWXNFOC-UHFFFAOYSA-NRAHZWNYVWXNFOC-UHFFFAOYSA-N
Sulfur dioxideSO2
dioxido de azufre
Dioxyde de soufre
Fermenticide liquid
Schwefeldioxid
SULFUR DIOXIDE, LIQUID
Sulfur superoxide
Sulfurous acid anhydride
Sulfurous anhydride
Sulfurous oxide
sulphur dioxide
UN 1079
DTXSID602967250-00-0WSFSSNUMVMOOMR-UHFFFAOYSA-NWSFSSNUMVMOOMR-UHFFFAOYSA-N
FormaldehydeFannoform
Floguard 1015
formaldehido
Formaldehyd
Formalin LM
Formalin Taisei
Formalith
Formic aldehyde
Lysoform
Methaldehyde
Methanal
Methyl aldehyde
Methylene oxide
Morbicid
NSC 298885
Optilyse
Oxomethane
Oxymethylene
Superlysoform
UN 1198
UN 2209
Caswell No. 465
EINECS 200-001-8
EPA Pesticide Chemical Code 043001
Formaldehyde, gas
Formalin 40
NCI-C02799
RCRA waste number U122
Aldehyd mravenci
Aldehyde formique
Aldeide formica
Formalin-loesungen
Oplossingen
UNII-1HG84L3525
Formalaz
DTXSID702063710102-43-9MWUXSHHQAYIFBG-UHFFFAOYSA-NMWUXSHHQAYIFBG-UHFFFAOYSA-N
Nitric oxideNitric oxide (NO)
Amidogen, oxo-
monoxido de nitrogeno
Monoxyde d'azote
Nitric oxide trimer
Nitrogen monooxide
nitrogen monoxide
Nitrogen(II) oxide
Nitrosyl radical
Oxido nitrico
Stickstoffmonoxid
UN 1660
DTXSID102093860-35-5DLFVBJFMPXGRIB-UHFFFAOYSA-NDLFVBJFMPXGRIB-UHFFFAOYSA-N
AcetamideAcetamid
acetamida
Acetic acid amide
Acetimidic acid
Ethanamide
Ethanimidic acid
Methanecarboxamide
NSC 25945
DTXSID7020005103-90-2RZVAJINKPMORJF-UHFFFAOYSA-NRZVAJINKPMORJF-UHFFFAOYSA-N
Acetaminophen4-Acetamidophenol
APAP
Paracetamol
4-hydroxyacetanilide
Acetamide, N-(4-hydroxyphenyl)-
4-(Acetylamino)phenol
4-(N-Acetylamino)phenol
4-Acetaminophenol
4'-Hydroxyacetanilide
Abensanil
Acetagesic
Acetalgin
ACETAMIDE, N-(4-HYDROXYPHENYL)
Acetaminofen
Acetanilide, 4'-hydroxy-
ACETANILIDE, 4-HYDROXY-
Algotropyl
Alvedon
Anaflon
Apamide
Banesin
Ben-u-ron
Bickie-mol
Biocetamol
Cetadol
Citramon P
Claratal
Clixodyne
Dafalgan
Daphalgan
Dial-a-gesic
Disprol
Doliprane
Dolprone
Dymadon
Efferalgan
Endophy
Febrilex
Febrilix
Febro-Gesic
Febrolin
Fepanil
Finimal
Gattaphen T
Gelocatil
Gutte Enteric
Homoolan
Jin Gang
Lestemp
Liquagesic
Lonarid
Lyteca Syrup
Minoset
Momentum
N-(4-Hydroxyphenyl)acetamide
N-Acetyl-4-aminophenol
N-Acetyl-4-hydroxyaniline
N-Acetyl-p-aminophenol
Napafen
Naprinol
Nobedon
NSC 109028
NSC 3991
Ortensan
p-(Acetylamino)phenol
p-Aceaminophenol
Pacemol
p-Acetamidophenol
p-Acetoaminophen
P-ACETYLAMINOPHENOL
Paldesic
panadeine
Panadol
Panadol Actifast
Panadol Extend
Panaleve
Panasorb
Panodil
Paracetamol DC
Paracetamole
Parageniol
Paramol
Paraspen
Parelan
Pasolind N
Perfalgan
Phenaphen
Phendon
p-Hydroxyacetanilide
Prodafalgan
Puerxitong
Pyrinazine
Resfenol
Resprin
Rhodapop NCR
Salzone
Tabalgin
Tachipirina
Tempanal
Tralgon
Tylenol
TylolHot
Valadol
Valgesic
Vermidon
Vick Pyrena
DTXSID2020006968-81-0VGZSUPCWNCWDAN-UHFFFAOYSA-NVGZSUPCWNCWDAN-UHFFFAOYSA-N
AcetohexamideBenzenesulfonamide, 4-acetyl-N-[(cyclohexylamino)carbonyl]-
1-(p-Acetylbenzenesulfonyl)-3-cyclohexylurea
1-[(p-Acetylphenyl)sulfonyl]-3-cyclohexylurea
Acetohexamid
acetohexamida
Dimelin
Dimelor
Dymelor
Gamadiabet
Hypoglicil
Metaglucina
Minoral
N-(p-Acetylphenylsulfonyl)-N'-cyclohexylurea
Ordimel
Tsiklamid
Urea, 1-[(p-acetylphenyl)sulfonyl]-3-cyclohexyl-
DTXSID702000767-66-3HEDRZPFGACZZDS-UHFFFAOYSA-NHEDRZPFGACZZDS-UHFFFAOYSA-N
ChloroformTrichloromethane
Methane, trichloro-
CARBON TRICHLORIDE
Chloroforme
cloroformo
Formyl trichloride
Methane trichloride
Methane,trichloro-
NSC 77361
Trichloroform
UN 1888
DTXSID1020306110-00-9YLQBMQCUIZJEEH-UHFFFAOYSA-NYLQBMQCUIZJEEH-UHFFFAOYSA-N
FuranDivinylene oxide
furanne
Furfuran
Oxacyclopentadiene
Tetrole
UN 2389
DTXSID60206467429-90-5XAGFODPZIPBFFR-UHFFFAOYSA-NAZDRQVAHHNSJOQ-UHFFFAOYSA-N
AluminumAisin Metal Fiber
Al 050P-H24
ALC Fine
Alcan XI 1391
Almi-Paste SSP 303AR
Aloxal 3010
Alpaste 00-0506
Alpaste 0100M
Alpaste 0100MA
Alpaste 0100M-C
Alpaste 0200M
Alpaste 0200T
Alpaste 0230M
Alpaste 0230T
Alpaste 0241M
Alpaste 0300M
Alpaste 0500M
Alpaste 0539X
Alpaste 0620MS
Alpaste 0625TS
Alpaste 0638-70C
Alpaste 0700M
Alpaste 0780M
Alpaste 0900M
Alpaste 100M
Alpaste 100MS
Alpaste 100MSR
Alpaste 1100M
Alpaste 1100MA
Alpaste 1100N
Alpaste 1100NA
Alpaste 1109MA
Alpaste 1109MC
Alpaste 1200M
Alpaste 1200T
Alpaste 1260MS
Alpaste 1500MA
Alpaste 1700NL
Alpaste 1810YL
Alpaste 1830YL
Alpaste 1900M
Alpaste 1900XS
Alpaste 1950M
Alpaste 1950N
Alpaste 210N
Alpaste 2172EA
Alpaste 2173
Alpaste 240T
Alpaste 241M
Alpaste 417
Alpaste 46-046
Alpaste 4-621
Alpaste 4919
Alpaste 50-63
Alpaste 50-635
Alpaste 51-148B
Alpaste 51-231
Alpaste 5205N
Alpaste 5207N
Alpaste 52-509
Alpaste 52-568
Alpaste 5301N
Alpaste 5302N
Alpaste 53-119
Alpaste 5422NS
Alpaste 54-452
Alpaste 54-497
Alpaste 54-542
Alpaste 55-516
Alpaste 55-519
Alpaste 55-574
Alpaste 5620NS
Alpaste 5630NS
Alpaste 5640NS
Alpaste 56-501
Alpaste 5650NS
Alpaste 5653NS
Alpaste 5654NS
Alpaste 5680N
Alpaste 5680NS
Alpaste 60-600
Alpaste 60-760
Alpaste 60-768
Alpaste 62-356
Alpaste 6340NS
Alpaste 6370NS
Alpaste 6390NS
Alpaste 640NS
Alpaste 65-388
Alpaste 66NLB
Alpaste 710N
Alpaste 7130N
Alpaste 7160N
Alpaste 7160NS
Alpaste 725N
Alpaste 740NS
Alpaste 7430NS
Alpaste 7580NS
Alpaste 7620NS
Alpaste 7640NS
Alpaste 7670M
Alpaste 7670NS
Alpaste 7675NS
Alpaste 7679NS
Alpaste 7680N
Alpaste 7680NS
Alpaste 76840NS
Alpaste 7730N
Alpaste 7770N
Alpaste 7830N
Alpaste 8004
Alpaste 8080N
Alpaste 8260NAR
Alpaste 891K
Alpaste 91-0562
Alpaste 92-0592
Alpaste 93-0595
Alpaste 93-0647
Alpaste 94-2315
Alpaste 95-0570
Alpaste 96-0635
Alpaste 96-2104
Alpaste 97-0510
Alpaste 97-0534
Alpaste AW 520B
Alpaste AW 612
Alpaste AW 9800
Alpaste F 795
Alpaste FM 7680K
Alpaste FX 440
Alpaste FX 910
Alpaste FZ 0534
Alpaste FZU 40C
Alpaste G
Alpaste HR 8801
Alpaste HS 2
Alpaste J
Alpaste K 9800
Alpaste MC 666
Alpaste MC 707
Alpaste MF 20
Alpaste MG 01
Alpaste MG 1000
Alpaste MG 1300
Alpaste MG 500
Alpaste MG 600
Alpaste MH 6601
Alpaste MH 8801
Alpaste MH 9901
Alpaste MR 7000
Alpaste MR 9000
Alpaste MS 630
Alpaste N 1700NL
Alpaste NS 7670
Alpaste O 100N
Alpaste O 2130
Alpaste O 300M
Alpaste P 0100
Alpaste P 1950
Alpaste S
Alpaste SAP 110
Alpaste SAP 414P
Alpaste SAP 550N
Alpaste SCR 5070
Alpaste TCR 2020
Alpaste TCR 2060
Alpaste TCR 2070
Alpaste TCR 3010
Alpaste TCR 3030
Alpaste TCR 3040
Alpaste TCR 3130
Alpaste TD 200T
Alpaste UF 500
Alpaste WB 0230
Alpaste WD 500
Alpaste WJP-U 75C
Alpaste WX 0630
Alpaste WX 7830
Alpaste WXA 7640
Alpaste WXM 0630
Alpaste WXM 0650
Alpaste WXM 0660
Alpaste WXM 1415
Alpaste WXM 1440
Alpaste WXM 5422
Alpaste WXM 760b
Alpaste WXM 7640
Alpaste WXM 7675
Alpaste WXM-T 60B
Alpaste WXM-U 75
Alpaste WXM-U 75C
Altop X
Aluchrome Ultrafin Super
Alumat 1600
Alumet H 30
aluminio
Aluminium
Aluminium Flake
Aluminum 27
Aluminum atom
Aluminum element
Aluminum Flake PCF 7620
Aluminum granules
ALUMINUM METAL/GRANULE
ALUMINUM PASTE
ALUMINUM PIGMENT
ALUMINUM TURNINGS
Alumi-paste 640NS
Alumipaste 91-0562
Alumipaste 98-1822T
Alumipaste AW 620
Alumipaste CR 300
Alumipaste GX 180A
Alumipaste GX 201A
Alumipaste HR 7000
Alumipaste HR 850
Alumipaste MG 11
Alumipaste MH 8801
Aquamet NPW 2900
Aquapaste 205-5
Aquasilver LPW
Astroflake 40
Astroflake Black N 020
Astroflake Black N 070
Astroflake LG 40
Astroflake LG 70
Astroflake Silver N 040
Astroshine NJ 1600
Astroshine T 8990
Atomizalumi VA 200
C.I. PIGMENT METAL 1
Chromal IV
Chromal X
Decomet 1001/10
Decomet 2018/10
Decomet High Gloss Al 1002/10
Ecka AS 081
Eckart 9155
Eterna Brite 301-1
Eterna Brite 601-1
Eterna Brite 651-1
Eterna Brite EBP 251PA
Eterna Brite Primier 251PA
Ferro FX 53-038
Friend Color F 500GR-W
Friend Color F 500WT
Friend Color F 700RE-W
Friend Color F 701RE-W
Hi Print 60T
High Print 60T
Hisparkle HS 2
Hydro Paste 8726
Hydrolac WHH 2153
Hydrolan 3560
Hydrolux Reflexal 100
Hydroshine WS 1001
JISA 51010P
Kryal Z
Lansford 243
LE Sheet 800
Leafing Alpaste
LG-H Silver 25
Lunar Al-V 95
Metallux 161
Metallux 2154
Metallux 2192
Metalure
Metalure 55350
Metalure L 55350
Metalure L 59510
Metalure W 2001
Metapor
Metasheen 1800
Metasheen HR 0800
Metasheen KM 100
Metasheen KM 1000
Metasheen Slurry 1807
Metasheen Slurry 1811
Metasheen Slurry KM 100
Metax G
Metax S
Mirror Glow 1000
Mirror Glow 600
Mirrorsheen
Noral Aluminium
Noral Ink Grade Aluminium
Obron 10890
Offset FM 4500
Puratronic
Reflexal 145
Reynolds 400
Reynolds 4-301
Reynolds 4-591
Reynolds 667
SAP 260PW-HS
SAP-FM 4010
SBC 516-20Z
Scotchcal 7755SE
Serumekku
Setanium 50MIS-H8
Siberline ET 2025
Siberline ST 21030E1
Silvar A
Silver VT 522
Silverline SSP 353
Silvex 793-20C
Sparkle Silver 3141ST
Sparkle Silver 3500
Sparkle Silver 3641
Sparkle Silver 5000AR
Sparkle Silver 516AR
Sparkle Silver 5242AR
Sparkle Silver 5245AR
Sparkle Silver 5271AR
Sparkle Silver 5500
Sparkle Silver 5745
Sparkle Silver 7000AR
Sparkle Silver 7005AR
Sparkle Silver 7500
Sparkle Silver 960-25E1
Sparkle Silver E 1745AR
Sparkle Silver L 1526AR
Sparkle Silver Premier 751
Sparkle Silver SS 3130
Sparkle Silver SS 5242AR
Sparkle Silver SS 5588
Sparkle Silver SSP 132AR
Special PCR 507
Splendal 6001BG
Spota Mobil 801
SSP 760-20C
Stapa Aloxal PM 2010
Stapa Aloxal PM 3010
Stapa Aloxal PM 4010
Stapa Hydrolac BG 8n.1
Stapa Hydrolac BGH Chromal X
Stapa Hydrolac PM Chromal VIII
Stapa Hydrolac W 60NL
Stapa Hydrolac WH 16
Stapa Hydrolac WH 66NL
Stapa Hydrolux 2192
Stapa Hydrolux 8154
Stapa IL Hydrolan 2192-55900G
Stapa Metallic R 607
Stapa Metallux 1050
Stapa Metallux 211
Stapa Metallux 212
Stapa Metallux 2196
Stapa Metallux 274
Stapa Mobilux 181
Stapa Offset 3000
Stapa PV 10
Stapa VP 46432G
Starbrite 2100
Super Fine 18000
Super Fine 22000
Supramex 2022
Toyo Aluminum 02-0005
Toyo Aluminum 93-3040
Transmet K 102HE
Tufflake 3645
Tufflake 5843
UN 1396
US Aluminum 809
Valimet H 2
Valimet H 3
White Silver 7080N
White Silver 7130N
DTXSID30402737440-43-9BDOSMKKIYDKNTQ-UHFFFAOYSA-NBDOSMKKIYDKNTQ-UHFFFAOYSA-N
CadmiumCadimium
CADMIUM BLUE
CADMIUM, IN PLATTEN, STANGEN, BROCKEN,KOERNER
DTXSID10239407439-97-6QSHDDOUJBYECFT-UHFFFAOYSA-NQSHDDOUJBYECFT-UHFFFAOYSA-N
MercuryLiquid silver
Mercure
MERCURIC METAL TRIPLE DISTILLED
mercurio
Mercury element
Quecksilber
Quicksilver
UN 2024
UN 2809
DTXSID10241727440-61-1JFALSRSLKYAFGM-UHFFFAOYSA-NJFALSRSLKYAFGM-UHFFFAOYSA-N
UraniumUranium, isotope of mass 238
238U Element
UN 2979 (DOT)
Uranium I
DTXSID10425227440-38-2RQNWIZPPADIBDY-UHFFFAOYSA-NRQNWIZPPADIBDY-UHFFFAOYSA-N
ArsenicAs
Arsenic black
ARSENIC METAL
arsenico
Grey arsenic
UN 1558
DTXSID40238867440-22-4BQCADISMDOOEFD-UHFFFAOYSA-NBQCADISMDOOEFD-UHFFFAOYSA-N
SilverAg Nanopaste NPS-J 90
Ag Sphere 2
Ag-C-GS
Algaedyn
Arctic Silver 3
Argentum
Astroflake 5
Carey Lea silver
Colloidal silver
Dotite XA 208
Du Pont 4943
ECM 100AF4810
Enlight 600
Enlight silver plate 600
Epinall
Finesphere SVND 102
Fordel DC
FP 5369-502
Jelcon SH 1
Jungindai Takasago 300
KS (metal)
LCP 1-19SFS
Metz 3000-1
Nanomelt AGC-A
Nanomelt Ag-XA 301
Nanomelt Ag-XF 301
Nanomelt Ag-XF 301H
Nanopaste NPS-J 90
Perfect Silver
Puff Silver X 1200
RT 1710S-C1
SD (metal)
Shell Silver
Silbest E 20
Silbest F 20
Silbest J 18
Silbest TC 12
Silbest TC 20E
Silbest TC 25A
Silbest TCG 1
Silbest TCG 7
Silcoat AgC 103
Silcoat AgC 2011
Silcoat AgC 209
Silcoat AgC 2190
Silcoat AgC 222
Silcoat AgC 2411
Silcoat AgC 74T
Silcoat AgC-A
Silcoat AgC-AO
Silcoat AgC-B
Silcoat AgC-BO
Silcoat AgC-D
Silcoat AgC-G
Silcoat AgC-GS
Silcoat AgC-L
Silcoat AgC-O
Silcoat GS
Silcoat RF 200
Silflake 135
Silsphere 514
Silver atom
Silver element
Silver Flake 1
Silver Flake 25
Silver Flake 52
Silver Flake 7A
SILVER FLAKES
Silver metal
Silvest TCG 11N
Technic 299
Technic 450
Techno Alpha 175
DTXSID40243057439-96-5PWHULOQIROXLJO-UHFFFAOYSA-NPWHULOQIROXLJO-UHFFFAOYSA-N
ManganeseColloidal manganese
Cutaval
Manganese element
Manganese fulleride
Manganese metal alloy
Manganese-55
manganeso
DTXSID20241697440-02-0PXHVJJICTQNCMI-UHFFFAOYSA-NPXHVJJICTQNCMI-UHFFFAOYSA-N
NickelCarbonyl 255
Carbonyl Ni 123
Carbonyl Ni 283
Carbonyl Nickel 123
Carbonyl Nickel 283
Carbonyl Nickel 287
Cerac N 2003
CNS 10 Micron
Exmet 4 Ni X-4/0
Fibrex P
Incofoam
Nickel element
NICKEL ROUND ANODES
Nicrobraz LM:BNi 2
Ni-Flake 95
Novamet 123
Novamet 4SP
Novamet 4SP10
Novamet 525
Novamet CNS 400
Novamet HCA 1
Novamet NI 255
Raney nickel
Raney nickel 2800
UN 1325
UN 2881
DTXSID20209257440-66-6HCHKCACWOHOZIP-UHFFFAOYSA-NHCHKCACWOHOZIP-UHFFFAOYSA-N
ZincZn
Asarco L 15
C.I. Pigment Black 16
Merrillite
NC-Zinc
Rheinzink
Stapa TE Zinc AT
UF (metal)
UN 1436
Zinc dust
Zinc Dust 3
Zinc Dust 500 mesh
Zinc Dust LS 2
Zinc Dust MCS
Zinc Flakes GTT
ZINC METAL
ZINC MOSSY
ZINC STRIP
ZINC, MOSSY
Zincsalt GTT
DTXSID7035012GO:0031514motile ciliumHP:0012262Abnormal ciliary motilityVT:0001947mucociliary clearance traitVT:0002327respiratory function traitMP:0003674oxidative stress3occurrence2decreased1increasedAcrolein2017-08-15T09:55:592021-09-28T08:23:20Ozone2021-07-21T10:18:562021-09-28T08:26:52Cigarette smoke2021-06-24T07:10:582021-09-28T09:07:54Nitrogen dioxide2021-09-09T07:26:052021-09-28T08:54:40Diesel engine exhaust2021-08-06T08:41:152021-09-28T08:55:47Acetaldehyde2021-07-22T07:51:432021-07-22T07:51:43Nicotine2021-07-22T07:53:072021-07-22T07:53:07Sex hormone2021-09-28T07:57:442021-09-28T07:57:44Sulfur dioxide2021-07-22T09:49:342021-07-22T09:49:34Formaldehyde2021-07-22T09:51:102021-07-22T09:51:10PM102021-07-22T09:54:462021-07-22T09:54:46Nitric oxide2021-07-22T09:57:382021-07-22T09:57:38Acetaminophen2016-11-29T18:42:262016-11-29T18:42:26Chloroform2016-11-29T18:42:272016-11-29T18:42:27furan2020-05-01T14:35:222020-05-01T14:35:22Platinum2022-02-04T14:36:542022-02-04T14:36:54Aluminum2022-02-04T14:42:112022-02-04T14:42:11Cadmium2017-10-25T08:33:122017-10-25T08:33:12Mercury2016-11-29T18:42:192016-11-29T18:42:19Uranium2021-08-05T14:28:502021-08-05T14:28:50Arsenic2021-04-27T00:15:212021-04-27T00:15:21Silver 2022-02-03T11:20:112022-02-03T11:20:11Manganese2022-02-04T14:47:232022-02-04T14:47:23Nickel2022-02-04T14:47:592022-02-04T14:47:59Zinc2022-02-04T15:05:002022-02-04T15:05:00nanoparticles2016-12-21T09:40:062016-12-21T09:40:069606Homo sapiens10090Mus musculus10116Rattus norvegicus9986Oryctolagus cuniculusWCS_9913Bos taurus10141Cavia porcellus8400Lithobates catesbeianus9825Sus scrofa domesticus9940Ovis aries9612Canis lupusWCS_8400Rana catesbeianaWCS_9606humanWikiUser_26rodentsCilia Beat Frequency, DecreasedCBF, DecreasedCellular<p>Cohesive beating of cilia lining the upper and lower respiratory tract is critical for efficient MCC. CBF is influenced by several factors including changes in the physical and chemical properties of the ASL (especially the periciliary fluid), structural modulation in the cilia, concentration of cyclic nucleotides cAMP and cGMP, and intracellular calcium (Ca<sup>2+</sup>). Formation of cyclic nucleotides such as cGMP is mediated by nitric oxide (NO), which is released by an enzyme family of nitric oxide synthases (NOSs) when the substrate L-arginine (L-Arg) is transformed to L-citrulline. NO activates its receptor protein, soluble guanylate cyclase (sGC), which catalyzes formation of cGMP from guanosine triphosphate (GTP). cGMP then activates protein kinase G (PKG) which has been implicated in the regulation of CBF (Jiao et al., 2011; Li et al., 2000). NO-dependent stimulation of CBF has also been associated with an increase in cAMP-dependent protein kinase A (PKA) (Di Benedetto et al., 1991; Lansley et al., 1992; Salathe et al., 1993; Sanderson and Dirksen, 1989; Schmid et al., 2007; Sisson et al., 1999; Uzlaner and Priel, 1999). An increase in intracellular endogenous cAMP was observed after treatment with isobutyl-1-methylxanthine that also increased CBF (Tamaoki et al., 1989). cAMP accumulation in the airway cilia has been shown to be dependent on Ca2+–calmodulin-dependent PDE1A and indirectly regulates CBF (Kogiso et al., 2018). Increase in CBF after treatment with NO substrate, L-arginine and inhibition of CBF by a NOS inhibitor, N-omega-nitro-L-arginine methyl ester (L-NAME) further provides evidence for the role of NO in increasing CBF (Jiao J. et al., 2011; Sisson J. H., 1995; Uzlaner and Priel, 1999; Yang et al., 1997). <br />
Modulation of CBF is not always accompanied by changes in cAMP levels. PKC activators, phorbol 12-myristate 13-acetate and L-o~-dioctanoylglycerol have been shown to decrease CBF in a concentration- and time-dependent manner in rabbit tracheal epithelial cells (Kobayashi et al., 1989). CBF has been shown to decrease after exposure to inhaled oxidants such as cigarette smoke across different species. A study with 120 subjects showed a significant decrease in nasal CBF following exposure to tobacco smoke (Agius et al., 1998). Exposure to cigarette smoke extract lead to reduction in forskolin-induced CBF in human sinonasal epithelium (Cohen et al., 2009) and isoproterenol- and methacholine-induced CBF in human adenoid tissues (Wang et al., 2012). This decrease in CBF and unresponsiveness to beta-agonist stimulation occurs in parallel to PKC activation and has been shown to be dependent on the duration of exposure to cigarette smoke in mice (Simet et al., 2010). Normal human bronchial epithelial cells exposed to aerosolized nicotine showed decreased CFTR and BK conductance, impaired CBF, ASL volume, and decreased expression of FOXJ1 and KCNMA1 (Garcia-Arcos et al., 2016). <br />
A concentration-dependent decrease in CBF has been observed after treatment with aldehydes. For example inhibition of cilia ATPase activity was observed after treatment with acetaldehyde, in ciliated bovine bronchial epithelial cells (Sisson et al., 1991). Acrolein, an aldehyde in the gas phase of cigarette smoke, induced ciliostasis at high concentrations (> 1 mM), after 5 min of treatment, and cellular necrosis after 3 hr. However, at lower concentrations (from 0.5‒1 mM), acrolein transiently reduced the CBF to 4 Hz (Romet et al., 1990).<br />
</p>
<p>There is no standardized method for measuring CBF. Digital high-speed video imaging with a manual count of CBF in slow motion video play is the most commonly used method for CBF measurement (Kim et al., 2011; Peabody et al., 2018). Photometry and video-microscopy have been used to measure CBF in vitro and ex vivo, including in ciliated bovine bronchial epithelial cells (Allen-Gipson et al., 2011; Sisson et al., 2003; Uzlaner and Priel, 1999), normal human bronchial epithelial cells (Feriani et al., 2017), human nasal epithelial cells (Dimova et al., 2005; Min et al., 1999b), human nasal ciliated epithelium (nasal brushings) (Agius et al., 1998), and mouse tracheal rings (Simet et al., 2010).<br />
CBF measurement in vitro generally involves mounting the tissue at the air-liquid interface on a stage followed by microscopic analysis and acquisition of images and/or video recordings of beating cilia. For in vivo and ex vivo measurements, Doppler optical coherence tomography (D-OCT) can also be applied, a mesoscopic non-contact imaging modality that provides high-resolution tomographic images and detects micromotion simultaneously (Jing et al., 2017). D-OCT has been used to quantitatively measure CBF in ex vivo rabbit tracheal cultures (Lemieux et al., 2015).</p>
<p>Age-dependent decreases in CBF have been demonstrated in several species (e.g. guinea pigs, mice, and human) (Bailey et al., 2014; Grubb et al., 2016; Ho et al., 2001; Joki and Saano, 1997; Paul et al., 2013). In a study with 46 healthy subjects with a wide age distribution (mean 42, range 19–81 years), age was found to be negatively associated with airway clearance of inhaled 6-μm Teflon particles (Svartengren et al., 2005).</p>
<p>Female hormones, i.e. progesterone and estrogen, have been shown to have direct effect on CBF, i.e., progesterone reduces CBF, 17β-estradiol and progesterone receptor antagonists counteract progesterone effects, but estradiol alone has also been shown to have no effect on CBF. However, the mechanism by which these hormones modulate CBF is yet to be elucidated (Jain et al., 2012; Jia et al., 2011).</p>
UBERON:0000115lung epitheliumCL:0005012multi-ciliated epithelial cellModerateMixedHighAll life stagesHighHighModerateHighHighModerateHigh<p>Agius, A. M., L. A. Smallman, and A. L. Pahor (1998). Age, smoking and nasal ciliary beat frequency. Clin. Otolaryngol. Allied Sci. 23, 227-230.</p>
<p>Allen-Gipson, D.S., Blackburn, M.R., Schneider, D.J., Zhang, H., Bluitt, D.L., Jarrell, J.C., et al. (2011). Adenosine activation of A(2B) receptor(s) is essential for stimulated epithelial ciliary motility and clearance. Am. J. Physiol. Lung Cell. Mol. Physiol. 301, L171-L180.</p>
<p>Bailey, K.L., Bonasera, S.J., Wilderdyke, M., Hanisch, B.W., Pavlik, J.A., Devasure, J., et al. (2014). Aging causes a slowing in ciliary beat frequency, mediated by PKCε. Am. J. Physiol. Lung Cell. Mol. Physiol. 306, L584-L589.</p>
<p>Cohen, N.A., Zhang, S., Sharp, D.B., Tamashiro, E., Chen, B., Sorscher, E.J., et al. (2009). Cigarette smoke condensate inhibits transepithelial chloride transport and ciliary beat frequency. Laryngoscope 119, 2269-2274.</p>
<p>Di Benedetto, G., Manara-Shediac, F.S. and Mehta, A. (1991). Effect of cyclic AMP on ciliary activity of human respiratory epithelium. Eur. Respir. J. 4, 789-795.</p>
<p>Dimova, S., Maes, F., Brewster, M.E., Jorissen, M., Noppe, M. and Augustijns, P. (2005). High-speed digital imaging method for ciliary beat frequency measurement. J. Pharmacy Pharmacol 57, 521-526.</p>
<p>Feriani, L., Juenet, M., Fowler, C.J., Bruot, N., Chioccioli, M., Holland, S.M., et al. (2017). Assessing the Collective Dynamics of Motile Cilia in Cultures of Human Airway Cells by Multiscale DDM. Biophys. J. 113, 109-119.</p>
<p>Garcia-Arcos, I., Geraghty, P., Baumlin, N., Campos, M., Dabo, A.J., Jundi, B., et al. (2016). Chronic electronic cigarette exposure in mice induces features of COPD in a nicotine-dependent manner. Thorax 71, 1119-1129.</p>
<p>Gosepath, J., Schaefer, D., Brommer, C., Klimek, L., Amedee, R.G., and Mann, W.J. (2000). Subacute Effects of Ozone Exposure on Cultivated Human Respiratory Mucosa. Am. J. Rhinol. 14, 411-418. </p>
<p>Grubb, B.R., Livraghi-Butrico, A., Rogers, T.D., Yin, W., Button, B. and Ostrowski, L.E. (2016). Reduced mucociliary clearance in old mice is associated with a decrease in Muc5b mucin. Am. J. Physiol. Lung Cell. Mol. Physiol. 310, L860-L867.</p>
<p>Ho, J.C., Chan, K.N., Hu, W.H., Lam, W.K., Zheng, L., Tipoe, G.L., et al. (2001). The Effect of Aging on Nasal Mucociliary Clearance, Beat Frequency, and Ultrastructure of Respiratory Cilia. Am. J. Respir. Crit. Care Med. 163, 983-988.</p>
<p>Jain, R., Ray, J.M., Pan, J.-H. and Brody, S.L. (2012). Sex hormone-dependent regulation of cilia beat frequency in airway epithelium. Am. J. Respir. Crit. Care Med. 46, 446-453.</p>
<p>Jia, S., Zhang, X., He, D.Z., Segal, M., Berro, A., Gerson, T., et al., 2011. Expression and Function of a Novel Variant of Estrogen Receptor–α36 in Murine Airways. Am. J. Respir. Cell Mol. Biol. 45, 1084-1089.</p>
<p>Jiao, J., Wang, H., Lou, W., Jin, S., Fan, E., Li, Y., et al. (2011). Regulation of ciliary beat frequency by the nitric oxide signaling pathway in mouse nasal and tracheal epithelial cells. Exp. Cell Res. 317, 2548-2553.</p>
<p>Jing, J.C., Chen, J.J., Chou, L., Wong, B.J.F. and Chen, Z. (2017). Visualization and Detection of Ciliary Beating Pattern and Frequency in the Upper Airway using Phase Resolved Doppler Optical Coherence Tomography. Sci. Rep. 7, 8522-8522.</p>
<p>Joki, S. and Saano, V. (1997). Influence of ageing on ciliary beat frequency and on ciliary response to leukotriene D4 in guinea‐pig tracheal epithelium. Clin. Exp. Pharmacol. Physiol. 24, 166-169.</p>
<p>Kim, W., Han, T.H., Kim, H.J., Park, M.Y., Kim, K.S. and Park, R.W. (2011). An Automated Measurement of Ciliary Beating Frequency using a Combined Optical Flow and Peak Detection. J. Healthc. Inform. Res. 17, 111-119.</p>
<p>Knoll, M., Shaoulian, R., Magers, T. and Talbot, P. (1995). Ciliary beat frequency of hamster oviducts is decreased in vitro by exposure to solutions of mainstream and sidestream cigarette smoke. Biol. Reprod. 53, 29-37.</p>
<p>Kobayashi, K., Tamaoki, J., Sakai, N., Chiyotani, A. and Takizawa, T. (1989). Inhibition of ciliary activity by phorbol esters in rabbit tracheal epithelial cells. Lung 167, 277-284.</p>
<p>Kogiso, H., Hosogi, S., Ikeuchi, Y., Tanaka, S., Inui, T., Marunaka, Y., et al. (2018). [Ca(2+) ]i modulation of cAMP-stimulated ciliary beat frequency via PDE1 in airway ciliary cells of mice. Exp. Physiol. 103, 381-390.</p>
<p>Lansley, A.B., Sanderson, M.J. and Dirksen, E.R. (1992). Control of the beat cycle of respiratory tract cilia by Ca2+ and cAMP. Am. J. Physiol. 263, L232-242.</p>
<p>Lemieux, B.T., Chen, J.J., Jing, J., Chen, Z. and Wong, B.J.F. (2015). Measurement of ciliary beat frequency using Doppler optical coherence tomography. Int. Forum Allergy Rhinol. 5, 1048-1054.</p>
<p>Li, D., Shirakami, G., Zhan, X. and Johns, R.A. (2000). Regulation of ciliary beat frequency by the nitric oxide-cyclic guanosine monophosphate signaling pathway in rat airway epithelial cells. Am. J. Respir. Cell Mol. Biol. 23, 175-181.</p>
<p>Min, Y.-G., Ohyama, M., Lee, K.S., Rhee, C.-S., Oh, S.H., Sung, M.-W., et al. (1999). Effects of free radicals on ciliary movement in the human nasal epithelial cells. Auris Nasus Larynx 26, 159-163.</p>
<p>Paul, P., Johnson, P., Ramaswamy, P., Ramadoss, S., Geetha, B. and Subhashini, A.S. (2013). The Effect of Ageing on Nasal Mucociliary Clearance in Women: A Pilot Study. ISRN Pulmonology 2013, 5.</p>
<p>Peabody, J.E., Shei, R.-J., Bermingham, B.M., Phillips, S.E., Turner, B., Rowe, S.M., et al. (2018). Seeing cilia: imaging modalities for ciliary motion and clinical connections. Am. J. Physiol. Lung Cell. Mol. Physiol. 314, L909-L921.</p>
<p>Romet, S., Dubreuil, A., Baeza, A., Moreau, A., Schoevaert, D. and Marano, F. (1990). Respiratory tract epithelium in primary culture: Effects of. Toxicol. In Vitro 4, 399-402.</p>
<p>Salathe, M., Pratt, M.M. and Wanner, A. (1993). Cyclic AMP-dependent phosphorylation of a 26 kD axonemal protein in ovine cilia isolated from small tissue pieces. Am. J. Respir. Cell Mol. Biol. 9, 306-314.</p>
<p>Sanderson, M.J. and Dirksen, E.R. (1989). Mechanosensitive and beta-adrenergic control of the ciliary beat frequency of mammalian respiratory tract cells in culture. Am. Rev. Respir. Dis. 139, 432-440.</p>
<p>Schmid, A., Sutto, Z., Nlend, M.-C., Horvath, G., Schmid, N., Buck, J., et al. (2007). Soluble Adenylyl Cyclase Is Localized to Cilia and Contributes to Ciliary Beat Frequency Regulation via Production of cAMP. J. Gen. Physiol. 130, 99-109.</p>
<p>Schmid, A., Baumlin, N., Ivonnet, P., Dennis, J.S., Campos, M., Krick, S., et al. (2015). Roflumilast partially reverses smoke-induced mucociliary dysfunction. Respir. Res. 16, 135.</p>
<p>Simet, S.M., Sisson, J.H., Pavlik, J.A., Devasure, J.M., Boyer, C., Liu, X., et al. (2010). Long-term cigarette smoke exposure in a mouse model of ciliated epithelial cell function. Am. J. Respir. Cell Mol. Biol. 43, 635-640.</p>
<p>Sisson, J.H. (1995). Ethanol stimulates apparent nitric oxide-dependent ciliary beat frequency in bovine airway epithelial cells. Am. J. Physiol. 268, L596-600.</p>
<p>Sisson, J.H., May, K. and Wyatt, T.A. (1999). Nitric oxide-dependent ethanol stimulation of ciliary motility is linked to cAMP-dependent protein kinase (PKA) activation in bovine bronchial epithelium. Alcohol Clin. Exp. Res. 23, 1528-1533.</p>
<p>Sisson, J.H., Stoner, J., Ammons, B. and Wyatt, T. (2003). All‐digital image capture and whole‐field analysis of ciliary beat frequency. J. Microsc. 211, 103-111.</p>
<p>Sisson, J.H., Tuma, D.J. and Rennard, S.I. (1991). Acetaldehyde-mediated cilia dysfunction in bovine bronchial epithelial cells. Am. J. Physiol. 260, L29-36.</p>
<p>Svartengren, M., Falk, R. and Philipson, K. (2005). Long-term clearance from small airways decreases with age. Eur. Respir. J. 26, 609-615.</p>
<p>Tamaoki, J., Kondo, M. and Takizawa, T. (1989). Effect of cAMP on ciliary function in rabbit tracheal epithelial cells. J. Appl. Physiol. 66, 1035-1039.</p>
<p>Uzlaner, N. and Priel, Z. (1999). Interplay between the NO pathway and elevated [Ca(2+)](i) enhances ciliary activity in rabbit trachea. J. Physiol. 516, 179-190.</p>
<p>Wang, L.F., White, D.R., Andreoli, S.M., Mulligan, R.M., Discolo, C.M. and Schlosser, R.J. (2012). Cigarette smoke inhibits dynamic ciliary beat frequency in pediatric adenoid explants. Otolaryngol. Head Neck Surg. 146, 659-663.</p>
<p>Yaghi, A., Zaman, A., Cox, G. and Dolovich, M.B. (2012). Ciliary beating is depressed in nasal cilia from chronic obstructive pulmonary disease subjects. Respir. Med. 106, 1139-1147.</p>
<p>Yang, B., Schlosser, R.J. and Mccaffrey, T.V. (1997). Signal transduction pathways in modulation of ciliary beat frequency by methacholine. Ann. Otol. Rhinol. Laryngol. 106, 230-236.</p>
2021-07-19T10:19:362021-09-10T01:38:13Mucociliary Clearance, DecreasedMCC, DecreasedIndividual<p>In healthy adults, tracheal mucus movement varies from 4 to >20 mm/min (Stannard and O'Callaghan, 2006), whereas mucociliary clearance (MCC) in the small airways is slower due to the lower number of ciliated cells (fewer cilia) and their shorter length (Foster et al., 1980; Iravani, 1969; Wanner et al., 1996).<br />
Since optimal MCC is dependent in multiple factors, including cilia number and structure as well as ASL and mucus properties, any disturbances of these can lead to impaired MCC. While high humidity or infection can enhance MCC, long-term exposure to noxious substances (e.g. cigarette smoke) lead to decreased mucus clearance from the airways. In most instances this is reflected by decreased mucus transport rates or velocities.<br />
</p>
<p>In humans, MCC has been assessed traditionally following inhalation of radio-labeled particles such as 99Tcm-labeled polystyrene particles, resin particles or serum albumin and following their clearance at regular intervals by radioimaging using gamma cameras (Agnew et al., 1986; Kärjä et al., 1982). Taking into account inhalation volumes and flow rates, lung airflow, particle deposition and retention, clearance rates can be calculated and effects of e.g. drugs on MCC can be examined. Alternatively, since MCC occurs at a similar rate in the nose to that in trachea and bronchi (Andersen and Proctor, 1983; Rutland and Cole, 1981) and for ease of use, measurements of MCC can be restricted to that of nasal MCC only. Probably one of the simplest methods is the saccharin transit test (STT). For this test, a small particle of saccharin is placed behind the anterior end of the inferior turbinate. The saccharin will be transported by mucociliary action toward the nasopharynx, where its sweet taste is perceived. When MCC is impaired, saccharin transit times will increase, with a 10- to 20-minute delay being considered a clinical sign of decreased MCC. Using the same principle, the test can also be performed or complemented with dyes such as indigo carmine or methylene blue (Deborah and Prathibha, 2014).</p>
<p>In experimental animals, MCC has been evaluated by gamma-scintigraphy (Greiff et al., 1990; Hua et al., 2010; Read et al., 1992), fluorescence videography/fluoroscopy (in explanted tracheas etc.) (Grubb et al., 2016; Rogers et al., 2018), or by 3D-SPECT (Ortiz Belda et al., 2016). Direct observation of particle movement across airway epithelia to determine mucus velocity or transport rates by using a fiberoptic bronchoscope may be helpful when working in larger animals such as dogs (King, 1998).<br />
In vitro, freshly excised frog palate preparations have been used to assess cilia function and mucociliary transport by videomicroscopy (Macchione et al., 1995; Macchione et al., 1999; Trindade et al., 2007). Murine and human nasal, bronchial and small airway epithelial models grown at the air-liquid interface are also suitable in vitro test systems for determining mucus transport by tracing inert particle movement with a set-up similar to that used for assessing CBF (Benam et al., 2018; Fliegauf et al., 2013; Knowles and Boucher, 2002; Sears et al., 2015).<br />
</p>
HighMixedHighAll life stagesHighModerateModerateModerateModerateModerateModerate<p>Agnew, J., Sutton, P., Pavia, D. and Clarke, S. (1986). Radioaerosol assessment of mucociliary clearance: towards definition of a normal range. Brit. J. Radiol. 59, 147-151.</p>
<p>Allegra, L., Moavero, N., and Rampoldi, C. (1991). Ozone-induced impairment of mucociliary transport and its prevention with N-acetylcysteine. Am. J. Med. 91, S67-S71.</p>
<p>Andersen, I. and Proctor, D. (1983). Measurement of nasal mucociliary clearance. Eur. J. Respir. Dis. Suppl. 127, 37-40.</p>
<p>Baby, M.K., Muthu, P.K., Johnson, P., and Kannan, S. (2014). Effect of cigarette smoking on nasal mucociliary clearance: A comparative analysis using saccharin test. Lung India 31, 39-42. </p>
<p>Benam, K.H., Vladar, E.K., Janssen, W.J. and Evans, C.M. (2018). Mucociliary defense: emerging cellular, molecular, and animal models. Ann. Am. Thorac. Soc. 15, S210-S215.</p>
<p>Deborah, S. and Prathibha, K., 2014. Measurement of nasal mucociliary clearance. Clin. Res. Pulmonol. 2, 1019.</p>
<p>Dülger, S., Akdeniz, Ö., Solmaz, F., Şengören Dikiş, Ö., and Yildiz, T. (2018). Evaluation of nasal mucociliary clearance using saccharin test in smokers: A prospective study. Clin. Respir. J. 12, 1706-1710. </p>
<p>Fliegauf, M., Sonnen, A.F.P., Kremer, B. and Henneke, P. (2013). Mucociliary Clearance Defects in a Murine In Vitro Model of Pneumococcal Airway Infection. PloS ONE 8, e59925.</p>
<p>Fló-Neyret, C., Lorenzi-Filho, G., Macchione, M., Garcia, M.L.B. and Saldiva, P.H.N. (2001). Effects of formaldehyde on the frog's mucociliary epithelium as a surrogate to evaluate air pollution effects on the respiratory epithelium. Braz. J. Med. Biol. Res. 34, 639-643.</p>
<p>Foster, W., Langenback, E. and Bergofsky, E. (1980). Measurement of tracheal and bronchial mucus velocities in man: relation to lung clearance. J. Appl. Physiol. 48, 965-971.</p>
<p>Greiff, L., Wollmer, P., Erjefält, I., Pipkorn, U. and Persson, C. (1990). Clearance of 99mTc DTPA from guinea pig nasal, tracheobronchial, and bronchoalveolar airways. Thorax 45, 841-845.</p>
<p>Grubb, B.R., Livraghi-Butrico, A., Rogers, T.D., Yin, W., Button, B. and Ostrowski, L.E. (2016). Reduced mucociliary clearance in old mice is associated with a decrease in Muc5b mucin. Am. J. Physiol. Lung Cell. Mol. Physiol. 310, L860-L867.</p>
<p>Habesoglu, M., Demir, K., Yumusakhuylu, A.C., Sahin Yilmaz, A., and Oysu, C. (2012). Does passive smoking have an effect on nasal mucociliary clearance? Otolaryngol Head Neck Surg. 147, 152-156.</p>
<p>Hua, X., Zeman, K.L., Zhou, B., Hua, Q., Senior, B.A., Tilley, S.L., et al. (2010). Noninvasive real-time measurement of nasal mucociliary clearance in mice by pinhole gamma scintigraphy. J. Appl. Physiol. 108, 189-196.</p>
<p>Iravani, J. (1969). Zum Mechanismus der Ortsabhängigkeit der Flimmeraktivität im Bronchialbaum/Location-Dependent Activity of the Ciliary Movement in the Bronchial Tree and its Possible Mechanism. In: Habermann E. et al. (eds) Naunyn Schmiedebergs Archiv für Pharmakologie. Springer, Berlin, Heidelberg.</p>
<p>Kakinoki Y, Ohashi Y, Tanaka A, Washio Y, Yamada K, Nakai Y, Morimoto K. (1998). Nitrogen dioxide compromises defence functions of the airway epithelium. Acta Oto-Laryngol. 118, 221-226.</p>
<p>Kärjä, J., Nuutinen, J. and Karjalainen, P. (1982). Radioisotopic Method for Measurement of Nasal Mucociliary Activity. Arch. Otolaryngol. 108, 99-101.</p>
<p>King, M. (1998). Experimental models for studying mucociliary clearance. Eur. Respir. J. 11, 222-228.</p>
<p>Knorst, M.M., Kienast, K., Riechelmann, H., Müller-Quernheim, J. and Ferlinz, R. (1994). Effect of sulfur dioxide on mucociliary activity and ciliary beat frequency in guinea pig trachea. Int. Arch. Occup. Environm. Health 65, 325-328.</p>
<p>Knowles, M.R. and Boucher, R.C. (2002). Mucus clearance as a primary innate defense mechanism for mammalian airways. J. Clin. Invest. 109, 571-577.</p>
<p>Macchione, M., Guimarães, E., Saldiva, P. and Lorenzi-Filho, G. (1995). Methods for studying respiratory mucus and mucus clearance. Braz. J. Med. Biol Res. 28, 1347.</p>
<p>Macchione, M., Oliveira, A.P., Gallafrio, C.T., Muchão, F.P., Obara, M.T., Guimarães, E.T., et al. (1999). Acute effects of inhalable particles on the frog palate mucociliary epithelium. Environm. Health Persp. 107, 829-833.</p>
<p>Morgan, K., Patterson, D. and Gross, E. (1986). Responses of the nasal mucociliary apparatus of F-344 rats to formaldehyde gas. Toxicol. Appl. Pharmacol. 82, 1-13.</p>
<p>Morgan, K.T., Patterson, D.L. and Gross, E.A. (1984). Frog palate mucociliary apparatus: structure, function, and response to formaldehyde gas. Fund. Appl. Toxicol. 4, 58-68.</p>
<p>Ortiz Belda, J.L., Ortiz, A., Milara Payá, J., Armengot Carceller, M., Sanz García, C., Compañ Quilis, D., et al. (2016). Evaluation of Mucociliary Clearance by Three Dimension Micro-CT-SPECT in Guinea Pig: Role of Bitter Taste Agonists. Plos ONE 11, e0164399.</p>
<p>Pagliuca, G., Rosato, C., Martellucci, S., De Vincentiis, M., Greco, A., Fusconi, M., et al. (2015). Cytologic and functional alterations of nasal mucosa in smokers: temporary or permanent damage? Otolaryngol Head Neck Surg 152, 740-745.</p>
<p>Proença, M., Xavier, R.F., Ramos, D., Cavalheri, V., Pitta, F., and Ramos, E.C. (2011). Immediate and short term effects of smoking on nasal mucociliary clearance in smokers. Revista Portuguesa de Pneumologia (English Edition) 17), 172-176.</p>
<p>Read, R.C., Roberts, P., Munro, N., Rutman, A., Hastie, A., Shryock, T., et al. (1992). Effect of Pseudomonas aeruginosa rhamnolipids on mucociliary transport and ciliary beating. J. Appl. Physiol. 72, 2271-2277.</p>
<p>Rogers, T.D., Ostrowski, L.E., Livraghi-Butrico, A., Button, B. and Grubb, B.R., 2018. Mucociliary clearance in mice measured by tracking trans-tracheal fluorescence of nasally aerosolized beads. Sci. Rep. 8, 1-12.</p>
<p>Rutland, J. and Cole, P.J. (1981). Nasal mucociliary clearance and ciliary beat frequency in cystic fibrosis compared with sinusitis and bronchiectasis. Thorax 36, 654-658.</p>
<p>Sears, P.R., Yin, W.-N. and Ostrowski, L.E. (2015). Continuous mucociliary transport by primary human airway epithelial cells in vitro. Am. J. Physiol. Lung Cell. Mol. Physiol. 309, L99-L108.</p>
<p>Solak, I., Marakoglu, K., Pekgor, S., Kargin, N.C., Alataş, N., and Eryilmaz, M.A. (2018). Nasal mucociliary activity changes in smokers. Konuralp Med. J. 10, 269-275.</p>
<p>Stannard, W. and O'callaghan, C. (2006). Ciliary function and the role of cilia in clearance. J. Aerosol Med. 19, 110-115.</p>
<p>Trindade, S.H.K., De Mello Júnior, J.F., De Godoy Mion, O., Lorenzi-Filho, G., Macchione, M., Guimarães, E.T., et al. (2007). Methods for Studying Mucociliary Transport. Braz. J. Otorhinolaryngol. 73, 704-712.</p>
<p>Wanner, A., Salathe, M. and O'riordan, T.G. (1996). Mucociliary clearance in the airways. Am. J. Respir. Crit. Care Med. 154, 1868-1902.</p>
<p>Xavier, R.F., Ramos, D., Ito, J.T., Rodrigues, F.M., Bertolini, G.N., Macchione, M., et al. (2013). Effects of cigarette smoking intensity on the mucociliary clearance of active smokers. Respiration 86, 479-485. </p>
<p>Yadav, J., and Kaushik, G. (2014). K Ranga R. Passive smoking affects nasal mucociliary clearance. J. Indian Acad. Clin. Med. 15, 96-99.</p>
<p>Yeates, D.B., Katwala, S.P., Daugird, J., Daza, A.V. and Wong, L.B. (1997). Excitatory and inhibitory neural regulation of tracheal ciliary beat frequency (CBF) activated by ammonia vapour and SO2. Ann. Occup. Hyg. 41, 736-744.</p>
2021-07-19T10:21:382021-09-10T07:19:28Decrease, Lung functionDecreased lung functionIndividual<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Segoe UI",sans-serif">Lung function is a clinical term referring to the physiological functioning of the lungs, most often in association with the tests used to assess it. Lung function loss can be caused by acute or chronic exposure to airborne toxicants or by an intrinsic disease of the respiratory system. </span></span></span></span></p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Although signs of cellular injury are typically exhibited first in the nose and larynx, alveolar-capillary barrier breakdown may ultimately arise and result in local edema (Miller and Chang, 2003). Clinically, bronchoconstriction and hypoxia are seen in the acute phase, with affected subjects exhibiting shortness of breath (dyspnea) and low blood oxygen saturation, and with reduced lung function indices of airflow, lung volume and gas exchange (Hert and Albert, 1994; and How it is Measured or Detected;). When alveolar damage is extensive, the reduced lung function can develop into acute respiratory distress syndrome (ARDS). This severe compromise of lung function is reflected by decreased gas exchange indices (PaO<sub>2</sub>/FIO<sub>2</sub> ≤200 mmHg, due to hypoxemia and impaired excretion of carbon dioxide), increased pulmonary dead space and decreased respiratory compliance (Matthay et al., 2019). Acute inhalation exposures to chemical irritants such as ammonia, hydrogen chloride, nitrogen oxides and ozone typically cause local edema that manifests as dyspnea and hypoxia. In cases where a breakdown of the alveolar capillary function ensues, ARDS develops. ARDS has a particularly high risk of mortality, estimated to be 30-40% (Gorguner and Akgun, 2010; Matthay et al., 2018; Reilly et al., 2019).</span></span></span></span></span></p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Lung function decrease due to reduction in lung volume is seen in pulmonary fibrosis, which can be linked to chronic exposures to e.g. silica, asbestos, metals, agricultural and animal dusts (Meltzer and Noble, 2008; Cheresh et al., 2013; Cosgrove, 2015; Trethewey and Walters, 2018). Additionally. decreased lung function occurs in pleural disease, chest wall and neuromuscular disorders, because of obesity and following pneumectomy (Moore, 2012). Decreased lung function can also be a result of narrowing of the airways by inflammation and mucus plugging resulting in airflow limitation. Decreased lung function is a feature of obstructive pulmonary diseases (e.g. asthma, COPD) and linked to a multitude of causes, including chronic exposure to cigarette smoke, dust, metals, organic solvents, asbestos, pathogens or genetic factors. </span></span></span></span></span></p>
<p style="text-align:justify"> </p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Pulmonary function tests are a group of tests that evaluate several parameters indicative of lung size, air flow and gas exchange. Decreased lung function can manifest in different ways, and individual circumstances, including potential exposure scenarios, determine which test is used. The section outlines the tests used to evaluate lung function in humans (https://www.nhlbi.nih.gov/health-topics/pulmonary-function-tests, accessed 22 March 2021) and in experimental animals.</span></span></span></span></span></p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><strong><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Lung function tests used to evaluate human lung function</span></span></strong></span></span></span></p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">The most common (“gold standard”) lung function test in human subjects is spirometry. Spirometry results are primarily used for diagnostic purposes, e.g. to discriminate between obstructive and restrictive lung diseases, and for determining the degree of lung function impairment. Specific criteria for spirometry tests have been outlined in the American Thoracic Society (ATS) and the European Respiratory Society (ERS) Task Force guidelines (Graham et al., 2019). These guidelines consist of detailed recommendations for the preparation and conduct of the test, instruction of the person tested, as well as indications and contraindications, and are complemented by additional guidance documents on how to interpret and report the test results (Pellegrino et al., 2005; Culver et al., 2017).</span></span></span></span></span></p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Spirometry measures several different parameters during forceful exhalation, including:</span></span></span></span></span></p>
<ul>
<li style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Forced expiratory volume in 1 s (FEV1), the maximum volume of air that can forcibly be exhaled during the first second following maximal inhalation</span></span></span></span></span></li>
<li style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Forced vital capacity (FVC), the maximum volume of air that can forcibly be exhaled following maximal inhalation </span></span></span></span></span></li>
<li style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Vital capacity (VC), the maximum volume of air that can be exhaled when exhaling as fast as possible</span></span></span></span></span></li>
<li style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">FEV1/FVC ratio</span></span></span></span></span></li>
<li style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Peak expiratory flow (PEF), the maximal flow that can be exhaled when exhaling at a steady rate</span></span></span></span></span></li>
<li style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Forced expiratory flow, also known as mid-expiratory flow; the rates at 25%, 50% and 75% FVC are given</span></span></span></span></span></li>
<li style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Inspiratory vital capacity (IVC), the maximum volume of air that can be inhaled after a full expiration</span></span></span></span></span></li>
</ul>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">A reduced FEV1, with normal or reduced VC, normal or reduced FVC, and a reduced FEV1/FVC ratio are indices of airflow limitation, i.e., airway obstruction as seen in COPD (Moore, 2012). In contrast, airway restriction is demonstrated by a reduction in FVC, normal or increased FEV1/FVC ratio, a normal spirometry trace and potentially a high PEF (Moore, 2012).</span></span></span></span></span></p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Lung capacity or lung volumes can be measured using one of three basic techniques: 1) plethysmography, 2) nitrogen washout, or 3) helium dilution. Plethysmography consists of a series of sequential measurements in a body plethysmograph, starting with the measurement of functional residual capacity (FRC),</span></span> <span style="font-family:"Segoe UI",sans-serif"><span style="color:black">the volume of gas present in the lung at end-expiration during tidal breathing. Once the FRC is known, expiratory reserve volume (ERV; the volume of gas that can be maximally exhaled from the end-expiratory level during tidal breathing, i.e., the FRC), vital capacity (VC; the volume change at the mouth between the positions of full inspiration and complete expiration), and inspiratory capacity (IC; the maximum volume of air that can be inhaled from FRC) are determined, and total lung capacity (TLC;</span></span> <span style="font-family:"Segoe UI",sans-serif"><span style="color:black">the volume of gas in the lungs after maximal inspiration, or the sum of all volume compartments) and residual volume (RV; the volume of gas remaining in the lung after maximal exhalation) are calculated (Weinstock and McCannon, 2017).</span></span></span></span></span></p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">The other two techniques used to measure lung volumes—helium dilution and nitrogen washout—are based on the principle of conservation of mass: [initial gas concentration] x [initial volume of the system] = [final gas concentration] x [final volume of the system]. The nitrogen washout method is based on the fact that nitrogen is present in the air, at a relatively constant amount. The subject is given 100% oxygen to breathe, and the expired gas, which contains nitrogen in the lung at the beginning of the test, is collected. When no more nitrogen is noted in the expirate, the volume of air expired and the entire amount of nitrogen in that volume are measured, and the initial volume of the system (FRC) can be calculated. In the helium dilution method, a known volume and concentration of helium is inhaled by the subject. Helium, an inert gas that is not absorbed significantly from the lungs, is diluted in proportion to the lung volume to which it is added. The final concentration of helium is then measured and FRC calculated (Weinstock and McCannon, 2017).</span></span></span></span></span></p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Measurements of lung volumes in humans are technically more challenging than spirometry. However, they complement spirometry (which cannot determine lung volumes) and may be a preferred means of lung function assessment when subject compliance cannot be reasonably expected (e.g. in pediatric subjects) or where forced expiratory maneuvers are not possible (e.g. in patients with advanced pulmonary fibrosis). There are recommended standards for lung volume measurements and their interpretation in clinical practice, issued by the ATS/ERS Task Force (Wanger et al., 2005; Criée et al., 2011). </span></span></span></span></span></p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Finally, indices of gas exchange across the alveolar-capillary barrier are tested by diffusion capacity of carbon monoxide (DLCO) studies (also referred to as transfer capacity of carbon monoxide, TLCO). The principle of the test is the increased affinity of hemoglobin to preferentially bind carbon monoxide over oxygen (Weinstock and McCannon, 2017). Complementary to spirometry and lung volume measurements, DLCO provides information about the lung surface area available for gas diffusion. Therefore, it is sensitive to any structural changes affecting the alveoli, such as those accompanying emphysema, pulmonary fibrosis, pulmonary edema, and ARDS. Recommendations for the standardization of the test and its evaluation have been outlined by the ATS/ERS Task Force (Graham et al., 2017). An isolated reduction in DLCO with normal spirometry and in absence of anemia suggests an injury to the alveolar-capillary barrier, as for example seen in the presence of pulmonary emboli or in patients with pulmonary hypertension (Weinstock and McCannon, 2017; Lettieri et al., 2006; Seeger et al., 2013). Reduced DLCO together with airflow obstruction (i.e., reduced FEV1) indicates lung parenchymal damage and is commonly observed in smokers and in COPD patients (Matheson et al., 2007; Harvey et al., 2016), whereas reduced DLCO with airflow restriction is seen in patients with interstitial lung diseases (Dias et al., 2014; Kandhare et al., 2016).</span></span></span></span></span></p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><strong><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Lung function tests used to evaluate experimental animal lung function</span></span></strong></span></span></span></p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Because spirometry requires active participation and compliance of the subject, it is not commonly used in animal studies. However, specialized equipment such as the flexiVent system (SCIREQ<sup>®</sup>) are available for measuring FEV, FVC and PEF in anesthetized and tracheotomized small laboratory animals. Other techniques such as plethysmography or forced oscillation are increasingly preferred for lung function assessment in small laboratory animals (McGovern et al., 2013; Bates, 2017). </span></span></span></span></span></p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">In small laboratory animals, plethysmography can be used to determine respiratory physiology parameters (minute volume, respiratory rate, time of pause and time of break), lung volume and airway resistance of conscious animals. Both whole body and head-out plethysmography can be applied, although there is a preference for the latter in the context of inhalation toxicity studies, because of its higher accuracy and reliability (OECD, 2018a; Hoymann, 2012).</span></span></span></span></span></p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Gas diffusion tests are not frequently performed in animals, because reproducible samplings of alveolar gas are difficult and technically challenging (Reinhard et al., 2002; Fallica et al., 2011). Modifications to the procedure employed in humans have, however, open possibilities to obtain a human-equivalent DLCO measure or the diffusion factor for carbon monoxide (DFCO)—a variable closely related to DLCO, which can inform on potential structural changes in the lungs that have an effect on gas exchange indices (Takezawa et al., 1980; Dalbey et al., 1987; Fallica et al., 2011; Limjunyawong et al., 2015).</span></span></span></span></span></p>
<p> </p>
<p><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Pulmonary function tests reflect the physiological working of the lungs. Therefore, the AO is applicable to a variety of species, including (but not limited to) rodents, rabbits, pigs, cats, dogs, horses and humans, independent of life stage and gender.</span></span></span></span></span></p>
HighMixedHighAdultHigh<div style="text-align:justify">
<p>Ackermann-Liebrich, U., Leuenberger, P., Schwartz, J., Schindler, C., Monn, C., Bolognini, G., et al. (1997). Lung function and long term exposure to air pollutants in Switzerland. Study on Air Pollution and Lung Diseases in Adults (SAPALDIA) Team. Am. J. Resp. Crit. Care Med. 155, 122-129. </p>
<p>Adam, M., Schikowski, T., Carsin, A.E., Cai, Y., Jacquemin, B., Sanchez, M., et al. (2015). Adult lung function and long-term air pollution exposure. ESCAPE: a multicentre cohort study and meta-analysis. Eur. Resp. J. 45, 38-50.</p>
<p>Andersen, M.H.G., Frederiksen, M., Saber, A.T., Wils, R.S., Fonseca, A.S., Koponen, I.K., et al. (2019). Health effects of exposure to diesel exhaust in diesel-powered trains. Part. Fibre Toxicol. 16, 21.</p>
<p>Ashley, F., Kannel, W.B., Sorlie, P.D., and Masson, R. (1975). Pulmonary function: relation to aging, cigarette habit, and mortality: the Framingham Study. Ann. Int. Med. 82, 739-745.</p>
<p>Bates, J.H.T. (2017). CO</p>
<p>Bergstra, A.D., Brunekreef, B., and Burdorf, A. (2018). The effect of industry-related air pollution on lung function and respiratory symptoms in school children. Environm. Health 17, 30. </p>
<p>Baumgartner, K.B., Samet, J.M., Coultas, D.B., Stidley, C.A., Hunt, W.C., Colby, T.V., and J.A. Waldron (2000). Occupational and environmental risk factors for idiopathic pulmonary fibrosis: a multicenter case-control study. Collaborating Centers. Am. J. Epidemiol. 152, 307-315.</p>
<p>Broekema, M., ten Hacken, N.H., Volbeda, F., Lodewijk, M.E., Hylkema, M.N., Postma, D.S., et al. (2009). Airway epithelial changes in smokers but not in ex-smokers with asthma. Am. J. Resp. Crit. Care Med. 180, 1170-1178.</p>
<p>Celli, B. R. (2010). Predictors of mortality in COPD. Respir. Med. 104, 773-779.</p>
<p>Cheresh, P., Kim, S.J., Tulasiram, S., and D.W. Kamp (2013). Oxidative stress and pulmonary fibrosis. Biochim. Biophys. Acta, 1832, 1028–1040.</p>
<p>Cosgrove, M.P. (2015). Pulmonary fibrosis and exposure to steel welding fume. Occup. Med. 65, 706-712.</p>
<p>Criée, C.P., Sorichter, S., Smith, H.J., Kardos, P., Merget, R., Heise, D., Berdel, D., Köhler, D., Magnussen, H., Marek, W. and H. Mitfessel (2011). Body plethysmography–its principles and clinical use. Respir. Med. 105, 959-971.</p>
<p>Dalbey, W., Henry, M., Holmberg, R., Moneyhun, J., Schmoyer, R. and S. Lock (1987). Role of exposure parameters in toxicity of aerosolized diesel fuel in the rat. J. Appl. Toxicol. 7, 265-275.</p>
<p>Dias, O.M., Baldi, B.G., Costa, A.N., C.R. Carvalho (2014). Combined pulmonary fibrosis and emphysema: an increasingly recognized condition. J. Bras. Pneumol. 40, 304-312. </p>
<p>Fallica, J., Das, S., Horton, M., and W. Mitzner (2011). Application of carbon monoxide diffusing capacity in the mouse lung. J. Appl. Physiol. 110, 1455–1459.</p>
<p>Forbes, L.J., Kapetanakis, V., Rudnicka, A.R., Cook, D.G., Bush, T., Stedman, J.R., et al. (2009). Chronic exposure to outdoor air pollution and lung function in adults. Thorax 64, 657-663.</p>
<p>Gold, D.R., Wang, X., Wypij, D., Speizer, F.E., Ware, J.H., and Dockery, D.W. (1996). Effects of cigarette smoking on lung function in adolescent boys and girls. N. Engl. J. Med. 335, 931-937. </p>
<p>Gorguner, M., and M. Akgun (2010). Acute inhalation injury. Euras. J. Med. 42, 28–35.</p>
<p>Graham, B.L., Brusasco, V., Burgos, F., Cooper, B.G., Jensen, R., Kendrick, A., MacIntyre, N.R., Thompson, B.R. and J. Wanger (2017). 2017 ERS/ATS standards for single-breath carbon monoxide uptake in the lung. Eur. Respir. J. 49, 1600016.</p>
<p>Graham, B.L., Steenbruggen, I., Miller, M.R., Barjaktarevic, I.Z., Cooper, B.G., Hall, G.L., Hallstrand, T.S., Kaminsky, D.A., McCarthy, K., McCormack, M.C. and C.E. Oropez (2019). Standardization of spirometry 2019 update. An official American Thoracic Society and European Respiratory Society technical statement. Am. J. Respir. Crit. Care Med. 200, e70-e88.</p>
<p>Harvey, B.G., Strulovici-Barel, Y., Kaner, R.J., Sanders, A., Vincent, T.L., Mezey, J.G. and R.G. Crystal (2016). Progression to COPD in smokers with normal spirometry/low DLCO using different methods to determine normal levels. Eur. Respir. J. 47, 1888-1889.</p>
<p>Hert, R. and R.K. Albert (1994). Sequelae of the adult respiratory distress syndrome. Thorax 49, 8-13.</p>
<p>Hoymann, H.G. (2012). Lung function measurements in rodents in safety pharmacology studies. Front. Pharmacol. 3, 156.</p>
<p>Johnson, J. D., and W. M. Theurer (2014). A stepwise approach to the interpretation of pulmonary function tests. Am. Fam. Phys. 89, 359-366.</p>
<p>Kandhare, A.D., Mukherjee, A., Ghosh, P. and S.L. Bodhankar (2016). Efficacy of antioxidant in idiopathic pulmonary fibrosis: A systematic review and meta-analysis. EXCLI J. 15, 636.</p>
<p>Kim, C.S., Alexis, N.E., Rappold, A.G., Kehrl, H., Hazucha, M.J., Lay, J.C., et al. (2011). Lung function and inflammatory responses in healthy young adults exposed to 0.06 ppm ozone for 6.6 hours. Am. J. Respir. Crit. Care Med. 183, 1215-1221.</p>
<p>Kuperman, A.S., and Riker, J.B. (1973). The variable effect of smoking on pulmonary function. Chest 63, 655-660. </p>
<p>Lee, H. M., Liu, M. A., Barrett-Connor, E., and N. D. Wong (2014). Association of Lung Function with Coronary Heart Disease and Cardiovascular Disease Outcomes in Elderly: The Rancho Bernardo Study. Respir. Med. 108, 1779–1785.</p>
<p>Lettieri, C.J., Nathan, S.D., Barnett, S.D., Ahmad, S. and A.F. Shorr (2006). Prevalence and outcomes of pulmonary arterial hypertension in advanced idiopathic pulmonary fibrosis. Chest 129, 746-752.</p>
<p>Limjunyawong, N., Fallica, J., Ramakrishnan, A., Datta, K., Gabrielson, M., Horton, M., and W. Mitzner (2015). Phenotyping mouse pulmonary function in vivo with the lung diffusing capacity. JoVE 95, e52216.</p>
<p>Matheson, M.C., Raven, J., Johns, D.P., Abramson, M.J. and E.H. Walters (2007). Associations between reduced diffusing capacity and airflow obstruction in community-based subjects. Respir. Med. 101, 1730-1737.</p>
<p>Matthay, M.A., Zemans, R.L., Zimmerman, G.A., Arabi, Y.M., Beitler, J.R., Mercat, A., Herridge, M., Randolph, A.G. and C.S. Calfee (2019). Acute respiratory distress syndrome. Nature Reviews Disease Primers 5, 1-22.</p>
<p>McGovern, T.K., Robichaud, A., Fereydoonzad, L., Schuessler, T.F., and J.G. Martin (2013) Evaluation of respiratory system mechanics in mice using the forced oscillation technique. JoVE 75, e50172.</p>
<p>Meltzer, E.B., and P.W. Noble (2008). Idiopathic pulmonary fibrosis. Orphanet J. Rare Dis. 3, 8.</p>
<p>Miller, K. and A. Chang (2003). Acute inhalation injury. Emerg. Med. Clin. N. Am. 21, 533-557.</p>
<p>Miller, M.R., Crapo, R., Hankinson, J., Brusasco, V., Burgos, F., Casaburi, R., Coates, A., Enright, P., van der Grinten, C.M., and P. Gustafsson (2005a). General considerations for lung function testing. Eur. Respir. J. 26, 153-161.</p>
<p>Miller, M.R., Hankinson, J., Brusasco, V., Burgos, F., Casaburi, R., Coates, A., Crapo, R., Enright, P., van der Grinten, C., and P. Gustafsson (2005b). Standardisation of spirometry. Eur. Respir. J. 26, 319-338.</p>
<p>Moore, V.C. (2012). Spirometry: step by step. Breathe 8, 232-240.</p>
<p>OECD (2018a). OECD Guidance Document on Inhalation Toxicity Studies, GD 39.</p>
<p>OECD (2018b), Test No. 412: Subacute Inhalation Toxicity: 28-Day Study, OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris, https://doi.org/10.1787/9789264070783-en.</p>
<p>OECD (2018), Test No. 413: Subchronic Inhalation Toxicity: 90-day Study, OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris, https://doi.org/10.1787/9789264070806-en.</p>
<p>Park, Y., Ahn, C., and T.H. Kim (2021) Occupational and environmental risk factors of idiopathic pulmonary fibrosis: a systematic review and meta-analyses. Sci. Rep. 11, 4318.</p>
<p>Prada-Dacasa, P., Urpi, A., Sánchez-Benito, L., Bianchi, P., A. Quintana (2020). Measuring Breathing Patterns in Mice Using Whole-body Plethysmography. Bio. Protoc. 10, e3741.</p>
<p>Pelkonen, M., Notkola, I.-L., Nissinen, A., Tukiainen, H., and Koskela, H. (2006). Thirty-year cumulative incidence of chronic bronchitis and COPD in relation to 30-year pulmonary function and 40-year mortality: a follow-up in middle-aged rural men. Chest 130, 1129-1137.</p>
<p>Pellegrino, R., Viegi, G., Brusasco, V., Crapo, R., Burgos, F., Casaburi, R., Coates, A., van der Grinten, C., Gustafsson, P., and J. Hankinson (2005). Interpretative strategies for lung function tests. Eur. Respir. J. 26, 948-968.</p>
<p>Raghu, G., Remy-Jardin, M., Myers, J.L., Richeldi, L., Ryerson, C.J., Lederer, D.J., Behr, J., Cottin, V., Danoff, S.K., Morell, F., and K.R. Flaherty (2018). Diagnosis of idiopathic pulmonary fibrosis. An official ATS/ERS/JRS/ALAT clinical practice guideline. Am. J. Respir. Crit. Care Med. 198, e44-e68.</p>
<p>Reilly, J.P., Zhao, Z., Shashaty, M.G., Koyama, T., Christie, J.D., Lanken, P.N., Wang, C., Balmes, J.R., Matthay, M.A., Calfee, C.S. and L.B. Ware (2019). Low to moderate air pollutant exposure and acute respiratory distress syndrome after severe trauma. Am. J. Respir. Crit. Care Med. 199, 62-70.</p>
<p>Reinhard. C., Eder, G., Fuchs, H., Ziesenis, A., Heyder, J. and H. Schulz H (2002). Inbred strain variation in lung function. Mamm. Genome 13, 429-437.</p>
<p>RP: Measurement of lung function in small animals. J. Appl. Physiol. 123, 1039-1046.</p>
<p>Schikowski, T., Sugiri, D., Ranft, U., Gehring, U., Heinrich, J., Wichmann, H.E., et al. (2005). Long-term air pollution exposure and living close to busy roads are associated with COPD in women. Respir. Res. 6, 152. </p>
<p>Seeger, W., Adir, Y., Barberà, J.A., Champion, H., Coghlan, J.G., Cottin, V., De Marco, T., Galiè, N., Ghio, S., Gibbs, S. and F.J. Martinez (2013). Pulmonary hypertension in chronic lung diseases. J. Am. Coll. Cardiol. 62 Suppl. 25, D109-D116.</p>
<p>Sin, D. D., Wu, L., and S. P. Man (2005). The relationship between reduced lung function and cardiovascular mortality: a population-based study and a systematic review of the literature. Chest 127, 1952-1959.</p>
<p>Takezawa, J., Miller, F.J. and J.J. O'Neil (1980). Single-breath diffusing capacity and lung volumes in small laboratory mammals. J. Appl. Physiol. 48, 1052-1059.</p>
<p>Tantisuwat, A., and Thaveeratitham, P. (2014). Effects of smoking on chest expansion, lung function, and respiratory muscle strength of youths. J. Phys. Ther. Sci. 26, 167-170. </p>
<p>Trethewey, S. P., and G. I. Walters (2018). The Role of Occupational and Environmental Exposures in the Pathogenesis of Idiopathic Pulmonary Fibrosis: A Narrative Literature Review. Medicina (Kaunas, Lithuania) 54, 108.</p>
<p>Tsui, H.-C., Chen, C.-H., Wu, Y.-H., Chiang, H.-C., Chen, B.-Y., and Guo, Y.L. (2018). Lifetime exposure to particulate air pollutants is negatively associated with lung function in non-asthmatic children. Environ. Poll. 236, 953-961. </p>
<p>Vestbo, J., Anderson, W., Coxson, H.O., Crim, C., Dawber, F., Edwards, L., Hagan, G., Knobil, K., Lomas, D.A., MacNee, W. and E.K. Silverman (2008). Evaluation of COPD longitudinally to identify predictive surrogate end-points (ECLIPSE). Eur. Respir. J. 31, 869-73.</p>
<p>Wanger, J., Clausen, J.L., Coates, A., Pedersen, O.F., Brusasco, V., Burgos, F., Casaburi, R., Crapo, R., Enright, P., Van Der Grinten, C.P.M. and P. Gustafsson (2005). Standardisation of the measurement of lung volumes. Eur. Respir. J. 26, 511-522.</p>
<p>Weinstock, T, and J. McCannon (2017). Pulmonary Medicine. Pulmonary Function Testing. https://www.pulmonologyadvisor.com/home/decision-support-in-medicine/pulmonary-medicine/pulmonary-function-testing/ (accessed 22 March 2021). Decision Support in Medicine, LLC.</p>
<p>Wise, R. A. (2006). The value of forced expiratory volume in 1 second decline in the assessment of chronic obstructive pulmonary disease progression. Am. J. Med. 119, 4-11.</p>
<p>Zhang, L.P., Zhang, X., Duan, H.W., Meng, T., Niu, Y., Huang, C.F., et al. (2017). Long-term exposure to diesel engine exhaust induced lung function decline in a cross sectional study. Ind.l Health 55, 13-26.</p>
</div>
2016-11-29T18:41:312021-09-08T04:54:28Oxidative Stress Oxidative Stress Molecular<p style="text-align:justify">Oxidative stress is defined as an imbalance in the production of reactive oxygen species (ROS) and antioxidant defenses. High levels of oxidizing free radicals can be very damaging to cells and molecules within the cell (Pizzino et al., 2017; Sharifi-Rad et al., 2020; Jena et al., 2023). As a result, the cell has important defense mechanisms to protect itself from ROS. For example, Nrf2 is a transcription factor and master regulator of the oxidative stress response. During periods of oxidative stress, Nrf2-dependent changes in gene expression are important in regaining cellular homeostasis (Nguyen, et al. 2009) and can be used as indicators of the presence of oxidative stress in the cell.</p>
<p style="text-align:justify">In addition to the directly damaging actions of ROS, cellular oxidative stress also changes cellular activities on a molecular level. Redox sensitive proteins have altered physiology in the presence and absence of ROS, which is caused by the oxidation of sulfhydryls to disulfides (2SH àSS) on neighboring amino acids (Antelmann and Helmann 2011). Importantly Keap1, the negative regulator of Nrf2, is regulated in this manner (Itoh, et al. 2010).</p>
<p><span style="font-size:16px"><span style="background-color:white"><span style="color:#2f5597">ROS also undermine the mitochondrial defense system from oxidative damage. The antioxidant systems consist of superoxide dismutase, <span style="background-color:white">catalase, glutathione peroxidase and glutathione reductase, as well as antioxidants such as α-tocopherol and ubiquinol</span></span></span><span style="color:#2f5597">, or antioxidant vitamins and minerals including vitamin E, C, carotene, lutein, zeaxanthin, selenium, and zinc (Fletcher, 2010). The enzymes, vitamins and minerals catalyze the conversion of ROS to non-toxic molecules such as water and O<sub>2</sub></span><span style="background-color:white"><span style="color:#2f5597"><span style="background-color:white">. However, these antioxidant systems are not perfect and endogenous metabolic processes and/or exogenous oxidative influences can trigger cumulative oxidative injuries to the mitochondria, causing a decline in their functionality and efficiency, which further promotes cellular oxidative stress (</span></span></span></span><span style="color:#2f5597">Balasubramanian, 2000; Ganea & Harding, 2006; Guo et al., 2013; Karimi et al., 2017)<span style="font-size:16px"><span style="background-color:white"><span style="background-color:white">.</span></span></span></span></p>
<p><span style="color:#27ae60"><span style="font-size:18px"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Calibri",sans-serif"><span style="background-color:white">However, an emerging viewpoint suggests that ROS-induced modifications may not be as detrimental as previously thought, but rather contribute to signaling processes (Foyer et al., 2017). </span></span></span></span></span></span></p>
<p style="text-align:justify">Protection against oxidative stress is relevant for all tissues and organs, although some tissues may be more susceptible. For example, the brain possesses several key physiological features, such as high O2 utilization, high polyunsaturated fatty acids content, presence of autooxidable neurotransmitters, and low antioxidant defenses as compared to other organs, that make it highly susceptible to oxidative stress (Halliwell, 2006; Emerit and al., 2004; Frauenberger et al., 2016).</p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:12.0pt"><span style="color:#2f5597">Sources of ROS Production</span></span></strong></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:12.0pt"><span style="color:#2f5597">Direct Sources:</span></span></strong><span style="font-size:12.0pt"><span style="color:#2f5597"> Direct sources involve the deposition of energy onto water molecules, breaking them into active radical species. When ionizing radiation hits water, it breaks it into hydrogen (H*) and hydroxyl (OH*) radicals by destroying its bonds. The hydrogen will create hydroxyperoxyl free radicals (HO<sub>2</sub>*) if oxygen is available, which can then react with another of itself to form hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) and more O<sub>2</sub> (Elgazzar and Kazem, 2015). Antioxidant mechanisms are also affected by radiation, with catalase (CAT) and peroxidase (POD) levels rising as a result of exposure (Seen et al. 2018; Ahmad et al. 2021). </span></span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:12.0pt"><span style="color:#2f5597">Indirect Sources:</span></span></strong><span style="font-size:12.0pt"><span style="color:#2f5597"> An indirect source of ROS is the mitochondria, which is one of the primary producers in eukaryotic cells (Powers et al., 2008). As much as 2% of the electrons that should be going through the electron transport chain in the mitochondria escape, allowing them an opportunity to interact with surrounding structures. Electron-oxygen reactions result in free radical production, including the formation of hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) (Zhao et al., 2019). The electron transport chain, which also creates ROS, is activated by free adenosine diphosphate (ADP), O<sub>2</sub>, and inorganic phosphate (P<sub>i</sub>) (Hargreaves et al. 2020; Raimondi et al. 2020; Vargas-Mendoza et al. 2021). The first and third complexes of the transport chain are the most relevant to mammalian ROS production (Raimondi et al., 2020). The mitochondria have its own set of DNA and it is a prime target of oxidative damage (Guo et al., 2013). ROS are also produced through nicotinamide adenine dinucleotide phosphate oxidase (NOX) stimulation, an event commenced by angiotensin II, a product/effector of the renin-angiotensin system (Nguyen Dinh Cat et al. 2013; Forrester et al. 2018). Other ROS producers include xanthine oxidase, immune cells (macrophage, neutrophils, monocytes, and eosinophils), phospholipase A<sub>2</sub> (PLA<sub>2</sub>), monoamine oxidase (MAO), and carbon-based nanomaterials (Powers et al. 2008; Jacobsen et al. 2008; Vargas-Mendoza et al. 2021).</span></span></span></span></p>
<p><strong>Oxidative Stress. Direct measurement of ROS is difficult because ROS are unstable. The presence of ROS can be assayed indirectly by measurement of cellular antioxidants, or by ROS-dependent cellular damage.</strong><span style="color:#27ae60"> Listed below are common methods for detecting the KE, however there may be other comparable methods that are not listed</span></p>
<ul>
<li>Detection of ROS by chemiluminescence <span style="font-size:12px">(<span style="font-family:arial,helvetica,sans-serif">https://www.sciencedirect.com/science/article/abs/pii/S0165993606001683)</span></span></li>
<li>Detection of ROS by chemiluminescence is also described in OECD TG 495 to assess phototoxic potential.</li>
<li>Glutathione (GSH) depletion. GSH can be measured by assaying the ratio of reduced to oxidized glutathione (GSH:GSSG) using a commercially available kit (e.g., http://www.abcam.com/gshgssg-ratio-detection-assay-kit-fluorometric-green-ab138881.html). </li>
<li>TBARS. Oxidative damage to lipids can be measured by assaying for lipid peroxidation using TBARS (thiobarbituric acid reactive substances) using a commercially available kit. </li>
<li>8-oxo-dG. Oxidative damage to nucleic acids can be assayed by measuring 8-oxo-dG adducts (for which there are a number of ELISA based commercially available kits),or HPLC, described in Chepelev et al. (Chepelev, et al. 2015).</li>
</ul>
<p><strong>Molecular Biology: Nrf2. Nrf2’s transcriptional activity is controlled post-translationally by oxidation of Keap1. Assay for Nrf2 activity include:</strong></p>
<ul>
<li>Immunohistochemistry for increases in Nrf2 protein levels and translocation into the nucleus</li>
<li>Western blot for increased Nrf2 protein levels</li>
<li>Western blot of cytoplasmic and nuclear fractions to observe translocation of Nrf2 protein from the cytoplasm to the nucleus</li>
<li>qPCR of Nrf2 target genes (e.g., Nqo1, Hmox-1, Gcl, Gst, Prx, TrxR, Srxn), or by commercially available pathway-based qPCR array (e.g., oxidative stress array from SABiosciences)</li>
<li>Whole transcriptome profiling by microarray or RNA-seq followed by pathway analysis (in IPA, DAVID, metacore, etc.) for enrichment of the Nrf2 oxidative stress response pathway (e.g., Jackson et al. 2014)</li>
<li>OECD TG422D describes an ARE-Nrf2 Luciferase test method</li>
<li>In general, there are a variety of commercially available colorimetric or fluorescent kits for detecting Nrf2 activation</li>
</ul>
<p> </p>
<table border="1" cellpadding="1" cellspacing="1">
<tbody>
<tr>
<td><strong>Assay Type & Measured Content</strong></td>
<td><strong>Description</strong></td>
<td><strong>Dose Range Studied</strong></td>
<td>
<p><strong>Assay Characteristics </strong><strong>(Length / Ease of use/Accuracy)</strong></p>
</td>
</tr>
<tr>
<td>
<p><strong>ROS Formation in the Mitochondria assay</strong> (Shaki et al., 2012)</p>
</td>
<td>“The mitochondrial ROS measurement was performed flow cytometry using DCFH-DA. Briefly, isolated kidney mitochondria were incubated with UA (0, 50, 100 and 200 μM) in respiration buffer containing (0.32 mM sucrose, 10 mM Tris, 20 mM Mops, 50 μM EGTA, 0.5 mM MgCl2, 0.1 mM KH2PO4 and 5 mM sodium succinate) [32]. In the interval times of 5, 30 and 60 min following the UA addition, a sample was taken and DCFH-DA was added (final concentration, 10 μM) to mitochondria and was then incubated for 10 min. Uranyl acetate-induced ROS generation in isolated kidney mitochondria were determined through the flow cytometry (Partec, Deutschland) equipped with a 488-nm argon ion laser and supplied with the Flomax software and the signals were obtained using a 530-nm bandpass filter (FL-1 channel). Each determination is based on the mean fluorescence intensity of 15,000 counts.”</td>
<td>0, 50, 100 and 200 μM of Uranyl Acetate</td>
<td>
<p>Long/ Easy</p>
<p>High accuracy</p>
</td>
</tr>
<tr>
<td>
<p><strong>Mitochondrial Antioxidant Content Assay</strong> Measuring GSH content</p>
(Shaki et al., 2012)</td>
<td>“GSH content was determined using DTNB as the indicator and spectrophotometer method for the isolated mitochondria. The mitochondrial fractions (0.5 mg protein/ml) were incubated with various concentrations of uranyl acetate for 1 h at 30 °C and then 0.1 ml of mitochondrial fractions was added into 0.1 mol/l of phosphate buffers and 0.04% DTNB in a total volume of 3.0 ml (pH 7.4). The developed yellow color was read at 412 nm on a spectrophotometer (UV-1601 PC, Shimadzu, Japan). GSH content was expressed as μg/mg protein.”</td>
<td>
<p>0, 50, 100, or 200 <em>μ</em>M Uranyl Acetate</p>
</td>
<td> </td>
</tr>
<tr>
<td>
<p><strong>H<sub>2</sub>O<sub>2</sub> Production Assay</strong> Measuring H<sub>2</sub>O<sub>2</sub> Production in isolated mitochondria</p>
(Heyno et al., 2008)</td>
<td>“Effect of CdCl<sub>2</sub> and antimycin A (AA) on H<sub>2</sub>O<sub>2</sub> production in isolated mitochondria from potato. H<sub>2</sub>O<sub>2</sub> production was measured as scopoletin oxidation. Mitochondria were incubated for 30 min in the measuring buffer (see the Materials and Methods) containing 0.5 mM succinate as an electron donor and 0.2 µM mesoxalonitrile 3‐chlorophenylhydrazone (CCCP) as an uncoupler, 10 U horseradish peroxidase and 5 µM scopoletin.” (</td>
<td>
<p>0, 10, 30  <em>μ</em>M Cd<sup>2+</sup></p>
2  <em>μ</em>M<br />
antimycin A</td>
<td> </td>
</tr>
<tr>
<td>
<p><strong>Flow Cytometry ROS & Cell Viability</strong></p>
(Kruiderig et al., 1997)</td>
<td>“For determination of ROS, samples taken at the indicated time points were directly transferred to FACScan tubes. Dih123 (10 mM, final concentration) was added and cells were incubated at 37°C in a humidified atmosphere (95% air/5% CO2) for 10 min. At <em>t </em>5 9, propidium iodide (10 mM, final concentration) was added, and cells were analyzed by flow cytometry at 60 ml/min. Nonfluorescent Dih123 is cleaved by ROS to fluorescent R123 and detected by the FL1 detector as described above for Dc (Van de Water 1995)”</td>
<td> </td>
<td>
<p>Strong/easy</p>
medium</td>
</tr>
<tr>
<td>
<p><strong>DCFH-DA Assay</strong> Detection of hydrogen peroxide production (Yuan et al., 2016)</p>
</td>
<td>
<p>Intracellular ROS production was measured using DCFH-DA as a probe. Hydrogen peroxide oxidizes DCFH to DCF. The probe is hydrolyzed intracellularly to DCFH carboxylate anion. No direct reaction with H<sub>2</sub>O<sub>2 </sub>to form fluorescent production. </p>
</td>
<td>0-400 µM</td>
<td>
<p>Long/ Easy</p>
<p>High accuracy</p>
</td>
</tr>
<tr>
<td>
<p><strong>H2-DCF-DA Assay</strong> Detection of superoxide production (Thiebault et al., 2007)</p>
</td>
<td>This dye is a stable nonpolar compound which diffuses readily into the cells and yields H2-DCF. Intracellular OH or ONOO- react with H2-DCF when cells contain peroxides, to form the highly fluorescent compound DCF, which effluxes the cell. Fluorescence intensity of DCF is measured using a fluorescence spectrophotometer.</td>
<td>0–600 µM</td>
<td>
<p>Long/ Easy</p>
<p>High accuracy</p>
</td>
</tr>
<tr>
<td><strong>CM-H2DCFDA Assay</strong></td>
<td>**Come back and explain the flow cytometry determination of oxidative stress from Pan et al. (2009)**</td>
<td> </td>
<td> </td>
</tr>
</tbody>
</table>
<p>Direct Methods of Measurement</p>
<table cellspacing="0" class="Table" style="border-collapse:collapse; width:623px">
<tbody>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:141px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:12.0pt"><span style="color:#2f5597">Method of Measurement</span></span></strong> </span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:151px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:12.0pt"><span style="color:#2f5597">References</span></span></strong> </span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:255px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:12.0pt"><span style="color:#2f5597">Description</span></span></strong> </span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:76px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:12.0pt"><span style="color:#2f5597">OECD-Approved Assay</span></span></strong></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:141px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Chemiluminescence </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:151px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">(Lu, C. et al., 2006; </span></span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Griendling, K. K., et al., 2016)</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:255px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">ROS can induce electron transitions in molecules, leading to electronically excited products. When the electrons transition back to ground state, chemiluminescence is emitted and can be measured. Reagents such as uminol and lucigenin are commonly used to amplify the signal. </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:76px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">No</span></span></span></span></p>
<p> </p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:141px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Spectrophotometry </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:151px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">(Griendling, K. K., et al., 2016)</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:255px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">NO has a short half-life. However, if it has been reduced to nitrite (NO2-), stable azocompounds can be formed via the Griess Reaction, and further measured by spectrophotometry. </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:76px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">No</span></span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:141px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Direct or Spin Trapping-Based Electron Paramagnetic Resonance (EPR) Spectroscopy </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:151px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">(Griendling, K. K., et al., 2016)</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:255px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">The unpaired electrons (free radicals) found in ROS can be detected with EPR, and is known as electron paramagnetic resonance. A variety of spin traps can be used. </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:76px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">No</span></span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:141px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Nitroblue Tetrazolium Assay </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:151px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">(Griendling, K. K., et al., 2016)</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:255px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">The Nitroblue Tetrazolium assay is used to measure O</span></span><sub><span style="font-size:12.0pt"><span style="color:#2f5597">2</span></span></sub><span style="background-color:white"><span style="color:#2f5597">•</span></span><sup><span style="font-size:12.0pt"><span style="color:#2f5597">–</span></span></sup><span style="font-size:12.0pt"><span style="color:#2f5597"> levels. O</span></span><sub><span style="font-size:12.0pt"><span style="color:#2f5597">2</span></span></sub><span style="background-color:white"><span style="color:#2f5597">•</span></span><sup><span style="font-size:12.0pt"><span style="color:#2f5597">–</span></span></sup><span style="font-size:12.0pt"><span style="color:#2f5597"> reduces nitroblue tetrazolium (a yellow dye) to formazan (a blue dye), and can be measured at 620 nm. </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:76px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">No</span></span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:141px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Fluorescence analysis of dihydroethidium (DHE) or Hydrocyans </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:151px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">(Griendling, K. K., et al., 2016)</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:255px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Fluorescence analysis of DHE is used to measure O</span></span><sub><span style="font-size:12.0pt"><span style="color:#2f5597">2</span></span></sub><span style="background-color:white"><span style="color:#2f5597">•</span></span><sup><span style="font-size:12.0pt"><span style="color:#2f5597">–</span></span></sup><span style="font-size:12.0pt"><span style="color:#2f5597"> levels. O</span></span><sub><span style="font-size:12.0pt"><span style="color:#2f5597">2</span></span></sub><span style="background-color:white"><span style="color:#2f5597">•</span></span><sup><span style="font-size:12.0pt"><span style="color:#2f5597">–</span></span></sup><span style="font-size:12.0pt"><span style="color:#2f5597"> is reduced to O2 as DHE is oxidized to 2-hydroxyethidium, and this reaction can be measured by fluorescence. Similarly, hydrocyans can be oxidized by any ROS, and measured via fluorescence. </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:76px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">No</span></span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:141px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Amplex Red Assay </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:151px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">(Griendling, K. K., et al., 2016)</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:255px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Fluorescence analysis to measure extramitochondrial or extracellular H<sub>2</sub>O<sub>2</sub> levels. In the presence of horseradish peroxidase and H<sub>2</sub>O<sub>2</sub>, Amplex Red is oxidized to resorufin, a fluorescent molecule measurable by plate reader. </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:76px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">No</span></span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:141px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Dichlorodihydrofluorescein Diacetate (DCFH-DA) </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:151px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">(Griendling, K. K., et al., 2016)</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:255px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">An indirect fluorescence analysis to measure intracellular H<sub>2</sub>O<sub>2</sub> levels. H<sub>2</sub>O<sub>2</sub> interacts with peroxidase or heme proteins, which further react with DCFH, oxidizing it to dichlorofluorescein (DCF), a fluorescent product. </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:76px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">No</span></span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:141px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">HyPer Probe </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:151px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">(Griendling, K. K., et al., 2016)</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:255px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Fluorescent measurement of intracellular H<sub>2</sub>O<sub>2</sub> levels. HyPer is a genetically encoded fluorescent sensor that can be used for <em>in vivo</em> and<em> in situ </em>imaging. </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:76px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">No</span></span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:141px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Cytochrome c Reduction Assay </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:151px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">(Griendling, K. K., et al., 2016)</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:255px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">The cytochrome c reduction assay is used to measure O</span></span><sub><span style="font-size:12.0pt"><span style="color:#2f5597">2</span></span></sub><span style="background-color:white"><span style="color:#2f5597">•</span></span><sup><span style="font-size:12.0pt"><span style="color:#2f5597">–</span></span></sup><span style="font-size:12.0pt"><span style="color:#2f5597"> levels. O</span></span><sub><span style="font-size:12.0pt"><span style="color:#2f5597">2</span></span></sub><span style="background-color:white"><span style="color:#2f5597">•</span></span><sup><span style="font-size:12.0pt"><span style="color:#2f5597">–</span></span></sup><span style="font-size:12.0pt"><span style="color:#2f5597"> is reduced to O2 as ferricytochrome c is oxidized to ferrocytochrome c, and this reaction can be measured by an absorbance increase at 550 nm. </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:76px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">No</span></span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:141px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Proton-electron double-resonance imagine (PEDRI)</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:151px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">(Griendling, K. K., et al., 2016)</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:255px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">The redox state of tissue is detected through nuclear magnetic resonance/magnetic resonance imaging, with the use of a nitroxide spin probe or biradical molecule. </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:76px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">No</span></span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:141px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Glutathione (GSH) depletion </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:151px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">(Biesemann, N. et al., 2018) </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:255px">
<p><span style="font-size:11pt"><span style="background-color:white"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">A downstream target of the Nrf2 pathway is involved in GSH synthesis. As an indication of oxidation status, GSH can be measured by assaying the ratio of reduced to oxidized glutathione (GSH:GSSG) using a commercially available kit (e.g., </span></span><span style="color:#2f5597"><a href="http://www.abcam.com/gshgssg-ratio-detection-assay-kit-fluorometric-green-ab138881.html"><span style="font-size:12.0pt"><span style="color:#2f5597">http://www.abcam.com/gshgssg-ratio-detection-assay-kit-fluorometric-green-ab138881.html</span></span></a></span><span style="font-size:12.0pt"><span style="color:#2f5597">). </span></span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:76px">
<p><span style="font-size:11pt"><span style="background-color:white"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">No</span></span></span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:141px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Thiobarbituric acid reactive substances (TBARS) </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:151px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">(Griendling, K. K., et al., 2016)</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:255px">
<p><span style="font-size:11pt"><span style="background-color:white"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Oxidative damage to lipids can be measured by assaying for lipid peroxidation with TBARS using a commercially available kit. </span></span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:76px">
<p><span style="font-size:11pt"><span style="background-color:white"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">No</span></span></span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:141px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Protein oxidation (carbonylation)</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:151px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">(Azimzadeh et al., 2017; Azimzadeh etal., 2015; Ping et al., 2020)</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:255px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Can be determined with enzyme-linked immunosorbent assay (ELISA) or a commercial assay kit. Protein oxidation can indicate the level of oxidative stress.</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:76px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">No</span></span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:141px"><span style="color:#27ae60">Seahorse XFp Analyzer </span></td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:151px"><span style="color:#27ae60">Leung et al. 2018 </span></td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:255px"><span style="color:#27ae60">The Seahorse XFp Analyzer provides information on mitochondrial function, oxidative stress, and metabolic dysfunction of viable cells by measuring respiration (oxygen consumption rate; OCR) and extracellular pH (extracellular acidification rate; ECAR). </span></td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:76px"><span style="color:#27ae60">No </span></td>
</tr>
</tbody>
</table>
<p><span style="font-size:11pt"><span style="background-color:white"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:12.0pt"><span style="color:#2f5597">Molecular Biology:</span></span></strong><span style="font-size:12.0pt"><span style="color:#2f5597"> Nrf2. Nrf2’s transcriptional activity is controlled post-translationally by oxidation of Keap1. Assays for Nrf2 activity include: </span></span></span></span></span></p>
<table cellspacing="0" class="Table" style="border-collapse:collapse; width:623px">
<tbody>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:154px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:12.0pt"><span style="color:#2f5597">Method of Measurement</span></span></strong> </span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:139px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:12.0pt"><span style="color:#2f5597">References</span></span></strong> </span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:256px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:12.0pt"><span style="color:#2f5597">Description</span></span></strong> </span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:75px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:12.0pt"><span style="color:#2f5597">OECD-Approved Assay</span></span></strong></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:154px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Immunohistochemistry </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:139px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">(Amsen, D., de Visser, K. E., and Town, T., 2009)</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:256px">
<p><span style="font-size:11pt"><span style="background-color:white"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Immunohistochemistry for increases in Nrf2 protein levels and translocation into the nucleus </span></span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:75px">
<p style="text-align:center"><span style="font-size:11pt"><span style="background-color:white"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">No</span></span></span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:154px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Quantitative polymerase chain reaction (qPCR) </span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:139px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">(Forlenza et al., 2012)</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:256px">
<p><span style="font-size:11pt"><span style="background-color:white"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">qPCR of Nrf2 target genes (e.g., Nqo1, Hmox-1, Gcl, Gst, Prx, TrxR, Srxn), or by commercially available pathway-based qPCR array (e.g., oxidative stress array from SABiosciences) </span></span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:75px">
<p style="text-align:center"><span style="font-size:11pt"><span style="background-color:white"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">No</span></span></span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; height:46px; vertical-align:top; width:154px">
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Whole transcriptome profiling via microarray or via RNA-seq followed by a pathway analysis</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:46px; vertical-align:top; width:139px">
<p><span style="font-size:11pt"><span style="background-color:white"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">(Jackson, A. F. et al., 2014)</span></span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:46px; vertical-align:top; width:256px">
<p><span style="font-size:11pt"><span style="background-color:white"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">Whole transcriptome profiling by microarray or RNA-seq followed by pathway analysis (in IPA, DAVID, metacore, etc.) for enrichment of the Nrf2 oxidative stress response pathway</span></span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:46px; vertical-align:top; width:75px">
<p style="text-align:center"><span style="font-size:11pt"><span style="background-color:white"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="color:#2f5597">No</span></span></span></span></span></p>
</td>
</tr>
</tbody>
</table>
<p><span style="color:#27ae60"><strong>Taxonomic applicability: </strong>Occurrence of oxidative stress is not species specific. </span></p>
<p><span style="color:#27ae60"><strong>Life stage applicability:</strong> Occurrence of oxidative stress is not life stage specific. </span></p>
<p><span style="color:#27ae60"><strong>Sex applicability: </strong>Occurrence of oxidative stress is not sex specific. </span></p>
<p><span style="color:#27ae60"><strong>Evidence for perturbation by prototypic stressor:</strong> There is evidence of the increase of oxidative stress following perturbation from a variety of stressors including exposure to ionizing radiation and altered gravity (Bai et al., 2020; Ungvari et al., 2013; Zhang et al., 2009). </span></p>
HighMixedHighAll life stagesHighHigh<p style="margin-left:48px; text-align:left"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">Ahmad, S. et al. (2021), “60Co-γ Radiation Alters Developmental Stages of Zeugodacus cucurbitae (Diptera: Tephritidae) Through Apoptosis Pathways Gene Expression”, <em>Journal Insect Science,</em> Vol. 21/5, Oxford University Press, Oxford, </span><a href="https://doi.org/10.1093/jisesa/ieab080" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1093/jisesa/ieab080</a></span></span></p>
<p style="margin-left:48px; text-align:left"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif">Antelmann, H. and J. D. Helmann (2011), “Thiol-based redox switches and gene regulation.”, <em>Antioxidants & Redox Signaling</em>, Vol. 14/6, Mary Ann Leibert Inc., Larchmont, <a href="https://doi.org/10.1089/ars.2010.3400" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1089/ars.2010.3400</a></span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Amsen, D., de Visser, K. E., and Town, T. (2009), “Approaches to determine expression of inflammatory cytokines”, in <em>Inflammation and Cancer</em>, Humana Press, Totowa, </span></span><a href="https://doi.org/10.1007/978-1-59745-447-6_5" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#1155cc">https://doi.org/10.1007/978-1-59745-447-6_5</span></span></span></a> </span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">Azimzadeh, O. et al. (2015), “Integrative Proteomics and Targeted Transcriptomics Analyses in Cardiac Endothelial Cells Unravel Mechanisms of Long-Term Radiation-Induced Vascular Dysfunction”, <em>Journal of Proteome Research</em>, Vol. 14/2, American Chemical Society, Washington, </span></span></span><a href="https://doi.org/10.1021/pr501141b" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1021/pr501141b</span></span></a></span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">Azimzadeh, O. et al. (2017), “Proteome analysis of irradiated endothelial cells reveals persistent alteration in protein degradation and the RhoGDI and NO signalling pathways”, <em>International Journal of Radiation Biology</em>, Vol. 93/9, Informa, London, </span></span></span><a href="https://doi.org/10.1080/09553002.2017.1339332" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1080/09553002.2017.1339332</span></span></a></span></span></p>
<p style="margin-left:48px"><span style="color:#27ae60">Azzam, E. I. et al. (2012), “Ionizing radiation-induced metabolic oxidative stress and prolonged cell injury”, Cancer Letters, Vol. 327/1-2, Elsevier, Ireland, https://doi.org/10.1016/j.canlet.2011.12.012 </span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">Bai, J. et al. (2020), “Irradiation-induced senescence of bone marrow mesenchymal stem cells aggravates osteogenic differentiation dysfunction via paracrine signaling”, <em>American Journal of Physiology - Cell Physiology</em>, Vol. 318/5, American Physiological Society, Rockville, </span></span></span><a href="https://doi.org/10.1152/ajpcell.00520.2019." style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1152/ajpcell.00520.2019.</span></span></a></span></span></p>
<p style="margin-left:48px"><span style="font-family:Times New Roman,Times,serif">Balasubramanian, D (2000), “Ultraviolet radiation and cataract”, <em>Journal of ocular pharmacology and therapeutics</em>, Vol. 16/3, Mary Ann Liebert Inc., Larchmont, <a href="https://doi.org/10.1089/jop.2000.16.285.%22%20/t%20%22_blank" rel="noreferrer noopener" target="_blank">https://doi.org/10.1089/jop.2000.16.285.</a> </span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Biesemann, N. et al., (2018), “High Throughput Screening of Mitochondrial Bioenergetics in Human Differentiated Myotubes Identifies Novel Enhancers of Muscle Performance in Aged Mice”, <em>Scientific Reports, </em>Vol. 8/1,</span></span> <span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Nature Portfolio, London, </span></span><a href="https://doi.org/10.1038/s41598-018-27614-8" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1038/s41598-018-27614-8</span></span></a><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">. </span></span></span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">Elgazzar, A. and N. Kazem. (2015), “Chapter 23: Biological effects of ionizing radiation” in <em>The Pathophysiologic Basis of Nuclear Medicine</em>, Springer, New York, pp. 540-548</span></span></span></span></span></p>
<p style="margin-left:48px"><span style="font-family:Times New Roman,Times,serif">Fletcher, A. E (2010), “Free radicals, antioxidants and eye diseases: evidence from epidemiological studies on cataract and age-related macular degeneration”, <em>Ophthalmic Research</em>, Vol. 44, Karger International, Basel, <a href="https://doi.org/10.1159/000316476.%22%20/t%20%22_blank" rel="noreferrer noopener" target="_blank">https://doi.org/10.1159/000316476.</a> </span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Forlenza, M. et al. (2012), “The use of real-time quantitative PCR for the analysis of cytokine mRNA levels” in <em>Cytokine Protocols, </em>Springer, New York, </span></span><a href="https://doi.org/10.1007/978-1-61779-439-1_2" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1007/978-1-61779-439-1_2</span></span></a><strong> </strong></span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">Forrester, S.J. et al. </span></span></span><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">(2018), “Angiotensin II Signal Transduction: An Update on Mechanisms of Physiology and Pathophysiology”, <em>Physiological Reviews, </em>Vol. 98/3<strong>,</strong> American Physiological Society, Rockville, </span></span></span><a href="https://doi.org/10.1152/physrev.00038.201" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1152/physrev.00038.201</span></span></a></span></span></p>
<p style="margin-left:48px"><span style="color:#27ae60"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Foyer, C. H., A. V. Ruban, and G. Noctor (2017), “Viewing oxidative stress through the lens of oxidative signalling rather than damage”, Biochemical Journal, Vol. 474/6, Portland Press, England, https://doi.org/10.1042/BCJ20160814 </span></span></span></span></span></p>
<p style="margin-left:48px"><span style="font-family:Times New Roman,Times,serif">Ganea, E. and J. J. Harding (2006), “Glutathione-related enzymes and the eye”, <em>Current eye research</em>, Vol. 31/1, Informa, London, <a href="https://doi.org/10.1080/02713680500477347.%22%20/t%20%22_blank" rel="noreferrer noopener" target="_blank">https://doi.org/10.1080/02713680500477347.</a> </span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="color:#222222">Griendling, K. K. et al. (2016), “Measurement of reactive oxygen species, reactive nitrogen species, and redox-dependent sign</span></span></span></span><span style="font-size:12.0pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="color:black">aling in the cardiovascular system: a scientific statement from the American Heart Association”, <em>Circulation research</em>, Vol. 119/5, Lippincott Williams & Wilkins, Philadelphia, </span></span></span></span><a href="https://doi.org/10.1161/RES.0000000000000110" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1161/RES.0000000000000110</span></span></span></a> </span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#303030">Guo, C.</span></span></span><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"> et al. (2013), “Oxidative stress, mitochondrial damage and neurodegenerative diseases”, <em>Neural regeneration research</em>, Vol. 8/21, Publishing House of Neural Regeneration Research, China, </span></span><a href="https://doi.org/10.3969/j.issn.1673-5374.2013.21.009" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/<span style="background-color:white">10.3969/j.issn.1673-5374.2013.21.009</span></span></span></a></span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#222222">Hargreaves, M., and L. L. Spriet (2020), “Skeletal muscle energy metabolism during exercise.”, <em>Nature Metabolism</em>, Vol. 2, Nature Portfolio, London, </span></span></span><a href="https://doi.org/10.1038/s42255-020-0251-4" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1038/s42255-020-0251-4</span></span></a></span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Hladik, D. and S. Tapio (2016), “Effects of ionizing radiation on the mammalian brain”, <em>Mutation Research/Reviews in Mutation Research</em>, Vol. 770, Elsevier, Amsterdam, </span></span><a href="https://doi.org/10.1016/j.mrrev.2016.08.003" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1016/j.mrrev.2016.08.003</span></span></a></span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Itoh, K., J. Mimura and M. Yamamoto (2010), “Discovery of the negative regulator of Nrf2, Keap1: a historical overview”, <em>Antioxidants & Redox Signaling</em>, Vol. 13/11, Mary Ann Leibert Inc., Larchmont, </span></span><a href="https://doi.org/10.1089/ars.2010.3222" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1089/ars.2010.3222</span></span></a> </span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Jackson, A.F. et al. (2014), “Case study on the utility of hepatic global gene expression profiling in the risk assessment of the carcinogen furan.”, <em>Toxicology and Applied Pharmacology, </em>Vol. 274/11, Elsevier, Amsterdam, </span></span><a href="https://doi.org/10.1016/j.taap.2013.10.019" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1016/j.taap.2013.10.019</span></span></a></span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">Jacobsen, N.R. et al. </span></span></span><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">(2008), “Genotoxicity, cytotoxicity, and reactive oxygen species induced by single-walled carbon nanotubes and C<sub>60</sub> fullerenes in the FE1-Muta<sup>TM </sup>Mouse lung epithelial cells”, <em>Environmental and Molecular Mutagenesis,</em> Vol. 49/6, John Wiley & Sons, Inc., Hoboken, </span></span></span><a href="https://doi.org/10.1002/em.20406" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1002/em.20406</span></span></a></span></span></p>
<p style="margin-left:48px"><span style="font-family:Times New Roman,Times,serif">Karimi, N. et al. (2017), “Radioprotective effect of hesperidin on reducing oxidative stress in the lens tissue of rats”, <em>International Journal of Pharmaceutical Investigation</em>, Vol. 7/3, Phcog Net, Bengaluru, <a href="https://doi.org/10.4103/jphi.JPHI_60_17.%E2%80%AF" rel="noreferrer noopener" target="_blank">https://doi.org/10.4103/jphi.JPHI_60_17.</a></span></p>
<p style="margin-left:48px"><span style="color:#27ae60"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Leung, D.T.H., and Chu, S. (2018), “Measurement of Oxidative Stress: Mitochondrial Function Using the Seahorse System” In: Murthi, P., Vaillancourt, C. (eds) Preeclampsia. Methods in Molecular Biology, vol 1710. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7498-6_22 </span></span></span></span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Lu, C., G. Song, and J. Lin (2006), “Reactive oxygen species and their chemiluminescence-detection methods”, <em>TrAC Trends in Analytical Chemistry, </em>Vol. 25/10, Elsevier, Amsterdam, </span></span><a href="https://doi.org/10.1016/j.trac.2006.07.007" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1016/j.trac.2006.07.007</span></span></a></span></span></p>
<p style="margin-left:48px; text-align:left"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">Nguyen Dinh Cat, A. et al. (2013), “Angiotensin II, NADPH oxidase, and redox signaling in the vasculature”, <em>Antioxidants & redox signaling,</em> Vol. 19/10<strong>,</strong> </span><span style="background-color:white"><span style="color:black">Mary Ann Liebert, Larchmont, </span></span><a href="https://doi.org/10.1089/ars.2012.4641" style="color:#0563c1; text-decoration:underline"><span style="background-color:white">https://doi.org/10.1089/ars.2012.4641</span></a></span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">Ping, Z. et al. (2020), “Oxidative Stress in Radiation-Induced Cardiotoxicity”, <em>Oxidative Medicine and Cellular Longevity</em>, Vol. 2020, Hindawi, </span></span></span><a href="https://doi.org/10.1155/2020/3579143" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1155/2020/3579143</span></span></a></span></span></p>
<p style="margin-left:48px">Pizzino, G. et al. (2017) “Oxidative Stress: Harms and Benefits for Human Health.” Oxidative medicine and cellular longevity, Vol. 2017: 8416763, Hindawi, https://doi.org/10.1155/2017/8416763 </p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">Powers, S.K. and M.J. Jackson. (2008), “Exercise-Induced Oxidative Stress: Cellular Mechanisms and Impact on Muscle Force Production”, <em>Physiological Reviews,</em> Vol. 88/4<strong>,</strong> American Physiological Society, Rockville, </span></span></span><a href="https://doi.org/10.1152/physrev.00031.2007" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1152/physrev.00031.2007</span></span></a></span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">Raimondi, V., F. Ciccarese and V. Ciminale. (2020), “Oncogenic pathways and the electron transport chain: a dangeROS liason”, <em>British Journal of Cancer, </em>Vol. 122/2, Nature Portfolio, London, </span></span></span><a href="https://doi.org/10.1038/s41416-019-0651-y" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1038/s41416-019-0651-y</span></span></a></span></span></p>
<p style="margin-left:48px; text-align:left"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">Seen, S. and L. Tong. (2018), “Dry eye disease and oxidative stress”, <em>Acta Ophthalmologica,</em> Vol. 96/4<strong>,</strong> John Wiley & Sons, Inc., Hoboken, </span><a href="https://doi.org/10.1111/aos.13526" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1111/aos.13526</a></span></span></p>
<p style="margin-left:48px; text-align:left"><span style="font-size:1rem">Sharifi-Rad, M. et al. (2020), “Lifestyle, Oxidative Stress, and Antioxidants: Back and Forth in the Pathophysiology of Chronic Diseases.” Frontiers in physiology Vol. 11:694, https://doi.org/10.3389/fphys.2020.00694 </span></p>
<p style="margin-left:48px; text-align:left">Snezhkina, A. V. et al. (2019), “ROS Generation and Antioxidant Defense Systems in Normal and Malignant Cells.” Oxidative medicine and cellular longevity Vol. 2019: 6175804, https://doi.org/10.1155/2019/6175804 </p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">Ungvari, Z. et al. (2013), “Ionizing Radiation Promotes the Acquisition of a Senescence-Associated Secretory Phenotype and Impairs Angiogenic Capacity in Cerebromicrovascular Endothelial Cells: Role of Increased DNA Damage and Decreased DNA Repair Capacity in Microvascular Radiosensitivity”, <em>The Journals of Gerontology Series A: Biological Sciences and Medical Sciences</em>, Vol. 68/12, Oxford University Press, Oxford, </span></span></span><a href="https://doi.org/10.1093/gerona/glt057." style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1093/gerona/glt057.</span></span></a> </span></span></p>
<p style="margin-left:48px"> </p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">Vargas-Mendoza, N. et al. (2021), “Oxidative Stress, Mitochondrial Function and Adaptation to Exercise: New Perspectives in Nutrition”, <em>Life, </em>Vol. 11/11, Multidisciplinary Digital Publishing Institute, Basel, </span></span></span><a href="https://doi.org/10.3390/life11111269" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.3390/life11111269</span></span></a></span></span></p>
<p style="margin-left:48px"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif">Wang, H. et al. (2019), “Radiation-induced heart disease: a review of classification, mechanism and prevention”, <em>International Journal of Biological Sciences, </em>Vol. 15/10, Ivyspring International Publisher, Sydney, <a href="https://doi.org/10.7150/ijbs.35460" style="color:#0563c1; text-decoration:underline">https://doi.org/10.7150/ijbs.35460</a> </span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#212121">Zhang, R. et al. (2009), “Blockade of AT1 receptor partially restores vasoreactivity, NOS expression, and superoxide levels in cerebral and carotid arteries of hindlimb unweighting rats”, <em>Journal of applied physiology</em>, Vol. 106/1, American Physiological Society, Rockville, </span></span></span><a href="https://doi.org/10.1152/japplphysiol.01278.2007" style="color:#0563c1; text-decoration:underline"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">https://doi.org/10.1152/japplphysiol.01278.2007</span></span></a><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#212121">.</span></span></span></span></span></p>
<p style="margin-left:48px"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">Zhao, R. Z. et al. (2019), “Mitochondrial electron transport chain, ROS generation and uncoupling”, <em>International journal of molecular medicine</em>, Vol. 44/1, </span><span style="color:black">Spandidos</span><span style="background-color:white"><span style="color:black"> Publishing Ltd</span></span><span style="color:black">., Athens, </span><a href="https://doi.org/10.3892/ijmm.2019.4188" style="color:#0563c1; text-decoration:underline">https://doi.org/10.3892/ijmm.2019.4188</a></span></span></p>
2017-05-30T13:58:172024-03-08T12:28:0812b168a9-f234-47b8-8775-0d8cf7357e8becb5c98b-102d-4798-ab8d-bb594cc3f1e9<p>Because the lung interfaces with the external environment, it is frequently exposed to airborne oxidant gases and particulates, and thus prone to oxidant-mediated cellular damage (Ciencewicki et al., 2008). Oxidant stress—through the action of exogenous and endogenous free radicals, such as super oxides, hydroxyl radicals, and hydrogen peroxides—is a common factor in lung inflammation and various respiratory diseases. The presence of redox-sensitive proteins in motile cilia suggests that oxidant stresses may impact ciliary function negatively (Price and Sisson, 2019). Indeed, exposure of human or rodent ciliated airway epithelial cells to hydrogen peroxide, acetaldehyde, ozone or cigarette smoke—all of which are known to cause oxidative stress—decreases CBF in a dose- and time-dependent manner (Bayram et al., 1998; Burman and Martin, 1986; Gosepath et al., 2000; Hastie et al., 1990; Helleday et al., 1995; Kienast et al., 1994; Knorst et al., 1994a; Min et al., 1999; Simet et al., 2010).</p>
<p>Experimental studies in vitro have shown that exposure of ciliated respiratory cells directly or indirectly to sources of oxidative stress leads to decreased CBF (Burman and Martin, 1986; Wilson et al., 1987; Feldman et al., 1994; Yoshitsugu et al., 1995; Min et al., 1999), which can be reversed by treatment with antioxidants (Schmid et al., 2015). Cigarette smoke condensate, a known inducer of oxidative stress, also causes a decrease in CBF in vitro (Cohen et al., 2009), while, in human subjects exposed to different oxygen levels, oxygen stress causes a decrease in nasal CBF (Stanek et al., 1998).<br />
</p>
<p>One mode of antimicrobial defense in the airway epithelium is generation of free radicals by neutrophils and monocytes/macrophages. Some microbes have also been shown to produce oxidants in significant amounts, e.g. H<sub>2</sub>O<sub>2</sub> production by pneumococcus. Several studies have shown that oxidants, irrespective of the source (microbial or host-derived) inhibit ciliary function. Additionally, there is a large body of experimental evidence demonstrating that exposures to environmental oxidants, including volatile aldehydes, peroxides, sulfur dioxide, nitric dioxide and Diesel exhaust particles have a detrimental impact on ciliary function. Therefore, this KER is highly plausible.</p>
<p>Treatment with H<sub>2</sub>O<sub>2</sub> causes a dose-dependent decline in ciliary beat frequency (CBF) in tracheal rings from male Sprague-Dawley rats (Burman and Martin, 1986).</p>
<p>Exposure of nasal ciliated epithelial cells to enzymatically generated oxidants (xanthine/xanthine oxidase) was accompanied by ciliary slowing, which was maximal at the end of the experiment (4 h). After 4 h, the reduction in CBF was 37.4%. Catalase alone, or in combination with superoxide dismutase (SOD), completely protected the epithelial strips from oxidant-mediated ciliary dyskinesia. Exposure of respiratory epithelium to glucose/glucose oxidase resulted in similar effects to those obtained with the xanthine/xanthine oxidase system, with 38% and 57% CBF reduction after 4 h in systems containing 25 and 100 mU of glucose oxidase, respectively. Treatments with H<sub>2</sub>O<sub>2</sub> and HOCl at concentrations of ≥100 µM caused dose-dependent ciliary dyskinesia after 4 h (Feldman et al., 1994).</p>
<p>Marked ciliary slowing was observed after exposure of human nasal epithelial cells to free radicals within the first 5 min. There was a significant difference in CBF between experimental and control groups. Pretreatment with 300 U/mL SOD or with 5 mM 3-ABA prevented CBF changes. (Min et al., 1999).</p>
<p>Pyocyanin dissolved in PBS produced gradual slowing of CBF in strips of normal human nasal ciliated epithelium without recovery. Ciliary dyskinesia was observed only late in the experiments when ciliostasis and epithelial disruption were also noted. In contrast, 1-hydroxyphenazine produced rapid onset of ciliary slowing, dyskinesia, and immediate ciliostasis but also some recovery of ciliary beating during the experiment (Wilson et al., 1987).</p>
<p>Baseline CBF was analyzed in human sinonasal epithelial cells for 4 min (time 0) followed by administration of either cigarette smoke condensate (CSC; not further specified) or DMSO for 3 min. Forskolin was then administered to the apical surface and stimulated CBF measured. Following addition of forskolin, stimulated CBF was significantly decreased in the CSC-exposed group compared to DMSO controls (Cohen et al., 2009).</p>
<p>Treatment of fully differentiated normal human bronchial epithelial cells with 100 nM roflumilast raised the CBF at 1 h after exposure. Subsequent air or cigarette smoke exposure increased CBF significantly in roflumilast-treated cultures (Schmid et al., 2015).</p>
<p>In superficial mucosae of the inferior nasal turbinates from non-smoking healthy volunteers exposed to three different oxygen concentrations—21%, 60% and 95%—for 2 h, CBF dose-dependently increased. Extending exposure to extreme oxygen concentrations to 4 h, however, decreased CBF, which is indicative of “oxygen stress” (Stanek et al., 1998).</p>
<p>Within 2 min of exposure of human respiratory epithelial cell monolayers to 10 µM H<sub>2</sub>O<sub>2</sub>, a decrease in CBF was observed. All cilia stopped moving within 10 min without obvious surface structural change in the ciliated cells. Catalase significantly reduced the ciliotoxic effect of H<sub>2</sub>O<sub>2 </sub>(Yoshitsugu et al., 1995).</p>
<p>Several studies show that oxidants decrease CBF which can be reversed by addition of antioxidants, suggesting a direct effect. However, there is evidence suggesting that oxidant-mediated decreases in CBF cannot be prevented by addition of antioxidants. For example, a polycyanin-induced decrease in CBF in human nasal epithelium could be reversed by treatment with isobutylmethylxanthine and forskolin, both of which increase intracellular cAMP, and also by the cAMP analog dibutyryl cAMP, while antioxidants did not seem to have any effect on CBF (Kanthakumar et al., 1993). Like polycyanin, two other <em>P. aeruginosa</em> toxins, 1-hydroxyphenazine (1-HP) and rhamnolipid reduced CBF which was associated with a decrease in intracellular adenosine nucleotides (Kanthakumar et al., 1996). </p>
<p>Inconsistent with several studies, there are studies that suggest that exposure to cigarette smoke does not inhibit CBF. A study involving 56 human subjects (27 non-smokers and 29 smokers) showed no differences in CBF between the 2 groups. However, a decrease in nasal mucociliary clearance was observed in smokers who exhaled smoke through their noses (Stanley et al., 1986). </p>
<p>While several studies have shown age dependence of CBF, there is evidence that suggests otherwise (Agius et al., 1998). </p>
<p>Several studies in various species, including humans and rodents, provide evidence in support of this KER. The empirical evidence confirms both a dose- and time-dependence between the upstream KE/MIE and the downstream KE. Our quantitative understanding of this KER is therefore strong.</p>
Not SpecifiedMixedNot SpecifiedAll life stagesHighNot SpecifiedNot SpecifiedNot Specified<p>Age-dependent decreases in CBF have been demonstrated in several species (e.g. guinea pigs, mice, and human) (Bailey et al., 2014; Grubb et al., 2016; Ho et al., 2001; Joki and Saano, 1997; Paul et al., 2013). </p>
<p>Female hormones, i.e. progesterone and estrogen, have been shown to have direct effect on CBF, i.e., progesterone reduces CBF, 17β-estradiol and progesterone receptor antagonists counteract progesterone effects, but estradiol alone has also been shown to have no effect on CBF. However, the mechanism by which these hormones modulate CBF is yet to be elucidated (Jain et al., 2012; Jia et al., 2011).</p>
<p>Agius, A.M., Smallman, L.A., and Pahor, A.L. (1998). Age, smoking and nasal ciliary beat frequency. Clin. Otolaryngol. Allied Sci. 23, 227-230. </p>
<p>Bailey, K.L., Bonasera, S.J., Wilderdyke, M., Hanisch, B.W., Pavlik, J.A., Devasure, J., et al. (2014). Aging causes a slowing in ciliary beat frequency, mediated by PKCε. Am. J. Physiol. Lung Cell. Mol. Physiol. 306, L584-L589.</p>
<p>Bayram, H., Devalia, J.L., Khair, O.A., Abdelaziz, M.M., Sapsford, R.J., Sagai, M., et al. (1998). Comparison of ciliary activity and inflammatory mediator release from bronchial epithelial cells of nonatopic nonasthmatic subjects and atopic asthmatic patients and the effect of diesel exhaust particles in vitro. J. Allergy Clin. Immunol. 102, 771-782.</p>
<p>Burman, W.J., and Martin, W.J. (1986). Oxidant-Mediated Ciliary Dysfunction: Possible Role in Airway Disease. Chest 89, 410-413. </p>
<p>Ciencewicki, J., Trivedi, S., and Kleeberger, S.R. (2008). Oxidants and the pathogenesis of lung diseases. J. Allergy Clin. Immunol. 122, 456-470. </p>
<p>Cohen, N.A., Zhang, S., Sharp, D.B., Tamashiro, E., Chen, B., Sorscher, E.J., et al. (2009). Cigarette smoke condensate inhibits transepithelial chloride transport and ciliary beat frequency. Laryngoscope 119, 2269-2274.</p>
<p>Feldman, C., Anderson, R., Kanthakumar, K., Vargas, A., Cole, P.J., and Wilson, R. (1994). Oxidant-mediated ciliary dysfunction in human respiratory epithelium. Free Radic. Biol. Med. 17, 1-10. </p>
<p>Gosepath, J., Schaefer, D., Brommer, C., Klimek, L., Amedee, R.G., and Mann, W.J. (2000). Subacute effects of ozone exposure on cultivated human respiratory mucosa. Am. J. Rhinol. 14, 411-418.</p>
<p>Grubb, B.R., Livraghi-Butrico, A., Rogers, T.D., Yin, W., Button, B. and Ostrowski, L.E. (2016). Reduced mucociliary clearance in old mice is associated with a decrease in Muc5b mucin. Am. J. Physiol. Lung Cell. Mol. Physiol. 310, L860-L867.</p>
<p>Hastie, A.T., Patrick, H., and Fish, J.E. (1990). Inhibition and recovery of mammalian respiratory ciliary function after formaldehyde exposure. Toxicol. Appl. Pharmacol. 102, 282-291.</p>
<p>Helleday, R., Huberman, D., Blomberg, A., Stjernberg, N., and Sandstrom, T. (1995). Nitrogen dioxide exposure impairs the frequency of the mucociliary activity in healthy subjects. Eur. Respir. J. 8, 1664-1668.</p>
<p>Ho, J.C., Chan, K.N., Hu, W.H., Lam, W.K., Zheng, L., Tipoe, G.L., et al. (2001). The Effect of Aging on Nasal Mucociliary Clearance, Beat Frequency, and Ultrastructure of Respiratory Cilia. Am. J. Respir. Crit. Care Med. 163, 983-988.</p>
<p>Jain, R., Ray, J.M., Pan, J.-H. and Brody, S.L. (2012). Sex hormone-dependent regulation of cilia beat frequency in airway epithelium. Am. J. Respir. Crit. Care Med. 46, 446-453.</p>
<p>Jia, S., Zhang, X., He, D.Z., Segal, M., Berro, A., Gerson, T., et al., 2011. Expression and Function of a Novel Variant of Estrogen Receptor–α36 in Murine Airways. Am. J. Respir. Cell Mol. Biol. 45, 1084-1089.</p>
<p>Joki, S. and Saano, V. (1997). Influence of ageing on ciliary beat frequency and on ciliary response to leukotriene D4 in guinea‐pig tracheal epithelium. Clin. Exp. Pharmacol. Physiol. 24, 166-169.</p>
<p>Kanthakumar, K., Taylor, G., Cundell, D., Dowling, R., Johnson, M., Cole, P., et al. (1996). The effect of bacterial toxins on levels of intracellular adenosine nucleotides and human ciliary beat frequency. Pulm. Pharmacol. 9, 223-230.</p>
<p>Kanthakumar, K., Taylor, G., Tsang, K., Cundell, D., Rutman, A., Smith, S., et al. (1993). Mechanisms of action of Pseudomonas aeruginosa pyocyanin on human ciliary beat in vitro. Infect. Immun. 61, 2848-2853.</p>
<p>Kienast, K., Riechelmann, H., Knorst, M., Schlegel, J., Müller-Quernheim, J., Schellenbergt, J., et al. (1994). An experimental model for the exposure of human ciliated cells to sulfur dioxide at different concentrations. The clinical investigator 72, 215-219. </p>
<p>Knorst, M.M., Kienast, K., Riechelmann, H., Müller-Quernheim, J., and Ferlinz, R. (1994). Effect of sulfur dioxide on mucociliary activity and ciliary beat frequency in guinea pig trachea. Int. Arch. Occup. Environ. Health 65, 325-328. </p>
<p>Min, Y.-G., Ohyama, M., Lee, K.S., Rhee, C.-S., Oh, S.H., Sung, M.-W., et al. (1999). Effects of free radicals on ciliary movement in the human nasal epithelial cells. Auris Nasus Larynx 26, 159-163.</p>
<p>Paul, P., Johnson, P., Ramaswamy, P., Ramadoss, S., Geetha, B. and Subhashini, A.S. (2013). The Effect of Ageing on Nasal Mucociliary Clearance in Women: A Pilot Study. ISRN Pulmonology 2013, 5.</p>
<p>Price, M.E., and Sisson, J.H. (2019). Redox regulation of motile cilia in airway disease. Redox. Biol. 27, 101146-101146. </p>
<p>Schmid, A., Baumlin, N., Ivonnet, P., Dennis, J.S., Campos, M., Krick, S., et al. (2015). Roflumilast partially reverses smoke-induced mucociliary dysfunction. Respir. Res. 16, 135. </p>
<p>Simet, S.M., Sisson, J.H., Pavlik, J.A., DeVasure, J.M., Boyer, C., Liu, X., et al. (2010). Long-term cigarette smoke exposure in a mouse model of ciliated epithelial cell function. Am. J. Respir. Cell Mol. Biol. 43, 635-640.</p>
<p>Sisson, J.H., Tuma, D.J., and Rennard, S.I. (1991). Acetaldehyde-mediated cilia dysfunction in bovine bronchial epithelial cells. Am. J. Physiol. 260, L29-36. </p>
<p>Stanek, A., Brambrink, A., Latorre, F., Bender, B., and Kleemann, P. (1998). Effects of normobaric oxygen on ciliary beat frequency of human respiratory epithelium. Br. J. Anaesth. 80, 660-664.</p>
<p>Stanley, P., Wilson, R., Greenstone, M., MacWilliam, L., and Cole, P. (1986). Effect of cigarette smoking on nasal mucociliary clearance and ciliary beat frequency. Thorax 41, 519-523.</p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Van Goor, F., S. Hadida, P. D. Grootenhuis, B. Burton, D. Cao, T. Neuberger, A. Turnbull, A. Singh, J. Joubran, A. Hazlewood, J. Zhou, J. McCartney, V. Arumugam, C. Decker, J. Yang, C. Young, E. R. Olson, J. J. Wine, R. A. Frizzell, M. Ashlock, and P. Negulescu. 2009. "Rescue of CF airway epithelial cell function in vitro by a CFTR potentiator, VX-770." <em>Proc Natl Acad Sci U S A</em> 106 (44): 18825-30. <a href="https://doi.org/10.1073/pnas.0904709106" style="color:blue; text-decoration:underline">https://doi.org/10.1073/pnas.0904709106</a>.</span></span></p>
<p>Wilson, R., Pitt, T., Taylor, G., Watson, D., MacDermot, J., Sykes, D., et al. (1987). Pyocyanin and 1-hydroxyphenazine produced by Pseudomonas aeruginosa inhibit the beating of human respiratory cilia in vitro. J. Clin. Investig. 79, 221-229.</p>
<p>Yoshitsugu, M., Matsunaga, S., Hanamure, Y., Rautiainen, M., Ueno, K., Miyanohara, T., et al. (1995). Effects of oxygen radicals on ciliary motility in cultured human respiratory epithelial cells. Auris Nasus Larynx 22, 178-185.</p>
2021-08-02T03:05:242023-03-24T06:04:15ecb5c98b-102d-4798-ab8d-bb594cc3f1e9932f4980-6e58-40a1-bffc-0c8fc0c177e3<p>Synchronized ciliary action transports mucus from the distal lung to the mouth, where it is swallowed or expectorated (Munkholm and Mortensen, 2014). In addition to ASL and mucus properties, the speed of ciliary movement, and hence the effectiveness of mucociliary clearance (MCC), is dependent on ciliary amplitude and beat frequency (Rubin, 2002). CBF itself is influenced by several factors, including changes in the physical and chemical properties of the ASL (especially the periciliary fluid), structural modulation in the cilia, concentration of cyclic nucleotides cAMP and cGMP, and intracellular calcium (Ca<sup>2+</sup>). Aside from genetic defects leading to ciliopathies, there is ample evidence for prolonged exposure to noxious agents, such as cigarette smoke, nitrogen oxide and sulfur dioxide, playing a major role in decreasing CBF and hampering efficient MCC.</p>
<p>A decrease in CBF resulting from sulfur dioxide exposure reduced mucociliary clearance in dogs (Yeates et al., 1997) and mucociliary activity in guinea pig tracheas (Knorst et al., 1994). In rats, formaldehyde inhalation exposure resulted in lower numbers of ciliated cells, while ciliary activity and mucus flow rates were decreased in a dose and time-dependent manner (Morgan et al., 1986). In humans, CBF positively correlates with nasal mucociliary clearance time (Ho et al., 2001), and bronchiectasis patients have lower nasal CBF and slower mucociliary transport (MCT) (Rutland and Cole, 1981). Administration of nebulized CBF inhibitors and enhancers quantifiably decreased or increased mucociliary clearance, respectively (Boek et al., 1999; Boek et al., 2002). Increased CBF and MCT was also noted in human sinonasal epithelial cell cultures treated with Myrtol®, an essential oil distillate (Lai et al., 2014) and in sheep tracheas and human airway epithelial cultures subjected to temperature changes (Kilgour et al., 2004; Sears et al., 2015). Exposures of frog palate epithelia to formaldehyde and PM10 reduced MCC and mucociliary transport, but only formaldehyde-treated epithelia showed decreases in CBF (Morgan et al., 1984; Macchione et al., 1999; Fló-Neyret et al., 2001).<br />
Ex vivo treatment of sheep trachea with acetylcholine and epinephrine increased CBF, but only acetylcholine increased surface liquid velocity, while both parameters were decreased upon incubation with platelet-activating factor (Seybold et al., 1990). </p>
<p>Ciliary function and mucus transport are invariably linked to effective mucus transport along the mucociliary escalator (Bustamante-Marin and Ostrowski, 2017; Mall, 2008). Therefore, this KER is biologically plausible. </p>
<p>Studies in animal models of ciliopathies and in individuals with genetic disorders causing cilia defects demonstrate that absent or asynchronous cilia beating results in defective mucus clearance from the lungs, consequently leading to respiratory infections that may be chronic recurrent in nature and ultimately lead to declining lung function (Knowles et al., 2013; Munkholm and Mortensen, 2014; Tilley et al., 2015). Similarly, indirect effects of airway inflammation, caused for example by respiratory infections or allergies, are known to be responsible for changes in cilia beating and hence mucus clearance (Almeida-Reis et al., 2010; Hisamatsu and Nakajima, 2000; Maurer et al., 1982). Finally, airway epithelial injury following exposure to inhalation toxicants can also damage cilia and inhibit cilia function and thereby impair MCC (Iravani and Van As, 1972; Wanner et al., 1996).</p>
<p>A decrease in CBF resulting from sulfur dioxide exposure reduced mucociliary clearance in dogs (Yeates et al., 1997) and mucociliary activity in guinea pig tracheas (Knorst et al., 1994). In rats, formaldehyde inhalation exposure resulted in lower numbers of ciliated cells, while ciliary activity and mucus flow rates were decreased in a dose and time-dependent manner (Morgan et al., 1986). In humans, CBF positively correlates with nasal mucociliary clearance time (Ho et al., 2001), and bronchiectasis patients have lower nasal CBF and slower mucociliary transport (MCT) (Rutland and Cole, 1981). Administration of nebulized CBF inhibitors and enhancers quantifiably decreased or increased mucociliary clearance, respectively (Boek et al., 1999; Boek et al., 2002). Increased CBF and MCT was also noted in human sinonasal epithelial cell cultures treated with Myrtol®, an essential oil distillate (Lai et al., 2014) and in sheep tracheas and human airway epithelial cultures subjected to temperature changes (Kilgour et al., 2004; Sears et al., 2015). Exposures of frog palate epithelia to formaldehyde and PM10 reduced MCC and mucociliary transport, but only formaldehyde-treated epithelia showed decreases in CBF (Morgan et al., 1984; Macchione et al., 1999; Fló-Neyret et al., 2001).</p>
<p>The available evidence does not interrogate the direct relationship between CBF and MCC, but rather evaluates both outcomes in parallel. However, because of the intrinsic linkage of cilia function and MCC, we find the empirical evidence in support of this KER to be moderate.<br />
Ex vivo treatment of sheep trachea with acetylcholine and epinephrine increased CBF, but only acetylcholine increased surface liquid velocity, while both parameters were decreased upon incubation with platelet-activating factor (Seybold et al., 1990). </p>
<p>Although ciliary function is considered a primary determinant for effective MCC (Duchateau et al., 1985; Gizurarson, 2015), there is evidence that suggests that MCC can be impeded by other factors that do not affect CBF. For example, nasal CBF in cigarette smokers regularly exhaling through the nose was not significantly different from that of nonsmokers, although they exhibited significantly longer nasomuciliary clearance times compared to nonsmokers. Possible explanations offered for this discrepancy were a potential loss of cilia in the nasal epithelium or increased mucus viscoelasticity (Stanley et al., 1986). Similarly, formaldehyde exposure of rats resulted in decreased cilia numbers and slower mucus flow rates (Morgan KT et al., 1986). On the other hand, there are a number of pharmacological compounds that improve mucociliary clearance through reductions in mucus viscosity, but have no effect on CBF (Jiao and Zhang, 2019), or through increases in CBF, but have no effect on mucociliary clearance (Phillips et al., 1990).</p>
<p>There are several studies providing insights into the negative effect of inhalation exposures on CBF and MCC, that are in line with the current thinking on how these two KEs connect. Additionally, pharmacological studies demonstrated that stimulation of CBF typically results in stimulation of MCC. However, since most studies usually evaluated the KEs in parallel, and even though some results support both dose response and temporal sequence of the KEs, none of the available data affirms causal linkage between CBF and MCC. Our understanding of the evidence is therefore moderate. </p>
HighMixedHighAll life stagesHighNot SpecifiedNot SpecifiedNot SpecifiedNot SpecifiedNot Specified<p>Evidences for this KER are derived from studies carried out in dog, gunea pig, rat, frog, sheep, rabbit model systems as well as in human epithelial cell cultures. MCC and CBF were observed to decrease with age in several species (e.g. guinea pigs, mice, and human) (Bailey et al., 2014; Grubb et al., 2016; Ho et al., 2001; Joki and Saano, 1997; Paul et al., 2013; Yager et al., 1978), but evidence by (Agius et al., 1998) suggests that age does not have a major effect on CBF.</p>
<p>Agius, A.M., Smallman, L.A., and Pahor, A.L. (1998). Age, smoking and nasal ciliary beat frequency. Clin. Otolaryngol. Allied Sci. 23, 227-230. </p>
<p>Almeida-Reis, R., Toledo, A.C., Reis, F.G., Marques, R.H., Prado, C.M., Dolhnikoff, M., et al. (2010). Repeated stress reduces mucociliary clearance in animals with chronic allergic airway inflammation. Respir. Physiol. Neurobiol. 173, 79-85.</p>
<p>Bailey, K.L., Bonasera, S.J., Wilderdyke, M., Hanisch, B.W., Pavlik, J.A., DeVasure, J., et al. (2014). Aging causes a slowing in ciliary beat frequency, mediated by PKCε. Am. J. Physiol. Lung Cell. Mol. Physiol. 306, L584-L589. </p>
<p>Boek, W.M., Graamans, K., Natzijl, H., van Rijk, P.P., and Huizing, E.H. (2002). Nasal Mucociliary Transport: New Evidence for a Key Role of Ciliary Beat Frequency. Laryngoscope 112, 570-573. </p>
<p>Boek, W.M., Keleş, N., Graamans, K., and Huizing, E.H. (1999). Physiologic and hypertonic saline solutions impair ciliary activity in vitro. Laryngoscope 109, 396-399.</p>
<p>Bustamante-Marin, X.M. and Ostrowski, L.E. (2017). Cilia and Mucociliary Clearance. Cold Spring Harb. Persp. Biol. 9, a028241.</p>
<p>Duchateau, G.S., Merkus, F.W., Zuidema, J., and Graamans, K. (1985). Correlation between nasal ciliary beat frequency and mucus transport rate in volunteers. The Laryngoscope 95, 854-859.</p>
<p>Fló-Neyret, C., Lorenzi-Filho, G., Macchione, M., Garcia, M.L.B., and Saldiva, P.H.N. (2001). Effects of formaldehyde on the frog's mucociliary epithelium as a surrogate to evaluate air pollution effects on the respiratory epithelium. Braz. J. Med. Biol. Res. 34, 639-643.</p>
<p>Gizurarson, S. (2015). The effect of cilia and the mucociliary clearance on successful drug delivery. Biol. Pharmaceut. Bull. b14-00398.</p>
<p>Grubb, B.R., Livraghi-Butrico, A., Rogers, T.D., Yin, W., Button, B., and Ostrowski, L.E. (2016). Reduced mucociliary clearance in old mice is associated with a decrease in Muc5b mucin. Am. J. Physiol. Lung Cell. Mol. Physiol. 310, L860-L867.</p>
<p>Hisamatsu, K.-i., and Nakajima, M. (2000). Pranlukast protects leukotriene C4- and D4-induced epithelial cell impairment of the nasal mucosa in vitro. Life Sci. 67, 2767-2773. </p>
<p>Ho, J.C., Chan, K.N., Hu, W.H., Lam, W.K., Zheng, L., Tipoe, G.L., et al. (2001). The Effect of Aging on Nasal Mucociliary Clearance, Beat Frequency, and Ultrastructure of Respiratory Cilia. Am. J. Respir. Crit. Care Med. 163, 983-988. </p>
<p>Houtmeyers, E., Gosselink, R., Gayan-Ramirez, G., and Decramer, M. (1999). Regulation of mucociliary clearance in health and disease. Eur. Respir. J. 13, 1177-1188.</p>
<p>Iravani, J., and Van As, A. (1972). Mucus transport in the tracheobronchial tree of normal and bronchitic rats. J. Pathol. 106, 81-93.</p>
<p>Jiao, J., and Zhang, L. (2019). Influence of Intranasal Drugs on Human Nasal Mucociliary Clearance and Ciliary Beat Frequency. Allergy Asthma Immunol. Res. 11, 306-319. </p>
<p>Joki, S., and Saano, V. (1997). Influence of ageing on ciliary beat frequency and on ciliary response to leukotriene D4 in guinea‐pig tracheal epithelium. Clin. Exp. Pharmacol. Physiol. 24, 166-169.</p>
<p>Kakinoki, Y.O., Ayaki Tanaka, Yushi Washio, Koji Yamada, Yoshiaki Nakai, Kazuhiro Morimoto, Yasushi (1998). Nitrogen dioxide compromises defence functions of the airway epithelium. Acta Otolaryngol. 118, 221-226.</p>
<p>Kilgour, E., Rankin, N., Ryan, S., and Pack, R. (2004). Mucociliary function deteriorates in the clinical range of inspired air temperature and humidity. Intensive Care Med. 30, 1491-1494.</p>
<p>Knorst, M.M., Kienast, K., Riechelmann, H., Müller-Quernheim, J., and Ferlinz, R. (1994). Effect of sulfur dioxide on mucociliary activity and ciliary beat frequency in guinea pig trachea. Int. Arch. Occup. Environ. Health 65, 325-328.</p>
<p>Knowles, M.R., Daniels, L.A., Davis, S.D., Zariwala, M.A., and Leigh, M.W. (2013). Primary ciliary dyskinesia. Recent advances in diagnostics, genetics, and characterization of clinical disease. Am. J. Respir. Crit. Care Med. 188, 913-922.</p>
<p>Lai, Y., Dilidaer, D., Chen, B., Xu, G., Shi, J., Lee, R.J., et al. (2014). In vitro studies of a distillate of rectified essential oils on sinonasal components of mucociliary clearance. Am. J. Rhinol. Allergy 28, 244-248.</p>
<p>Macchione, M., Oliveira, A.P., Gallafrio, C.T., Muchão, F.P., Obara, M.T., Guimarães, E.T., et al. (1999). Acute effects of inhalable particles on the frog palate mucociliary epithelium. Environ. Health Perspect. 107), 829-833.</p>
<p>Mall, M.A. (2008). Role of cilia, mucus, and airway surface liquid in mucociliary dysfunction: lessons from mouse models. J. Aerosol Med. Pulm. Drug Delivery 21, 13-24.</p>
<p>Maurer, D., Sielczak, M., Oliver Jr, W., Abraham, W., and Wanner, A. (1982). Role of ciliary motility in acute allergic mucociliary dysfunction. J. Appl. Physiol. 52, 1018-1023.</p>
<p>Morgan, K., Patterson, D., and Gross, E. (1986). Responses of the nasal mucociliary apparatus of F-344 rats to formaldehyde gas. Toxicol. Appl. Pharmacol. 82, 1-13.</p>
<p>Munkholm, M., and Mortensen, J. (2014). Mucociliary clearance: pathophysiological aspects. Clin. Physiol. Funct. Imaging 34, 171-177.</p>
<p>Paul, P., Johnson, P., Ramaswamy, P., Ramadoss, S., Geetha, B., and Subhashini, A.S. (2013). The Effect of Ageing on Nasal Mucociliary Clearance in Women: A Pilot Study. ISRN Pulmonology 2013, 5. </p>
<p>Phillips, P.P., McCaffrey, T.V., and Kern, E.B. (1990). The in vivo and in vitro effect of phenylephrine (Neo Synephrine) on nasal ciliary beat frequency and mucociliary transport. Otolaryngology Head Neck Surg. 103, 558-565.</p>
<p>Rubin, B.K. (2007). Mucus structure and properties in cystic fibrosis. Paediatr. Respir. Rev. 8, 4-7. </p>
<p>Rutland, J., and Cole, P.J. (1981). Nasal mucociliary clearance and ciliary beat frequency in cystic fibrosis compared with sinusitis and bronchiectasis. Thorax 36, 654-658.</p>
<p>Sears, P.R., Yin, W.-N., and Ostrowski, L.E. (2015). Continuous mucociliary transport by primary human airway epithelial cells in vitro. Am. J. Physiol. Lung Cell. Mol. Physiol. 309, L99-L108. </p>
<p>Seybold, Z.V., Mariassy, A.T., Stroh, D., Kim, C.S., Gazeroglu, H., and Wanner, A. (1990). Mucociliary interaction in vitro: effects of physiological and inflammatory stimuli. J. Appl. Physiol. 68, 1421-1426. </p>
<p>Stanley, P., Wilson, R., Greenstone, M., MacWilliam, L., and Cole, P. (1986). Effect of cigarette smoking on nasal mucociliary clearance and ciliary beat frequency. Thorax 41, 519-523.</p>
<p>Tilley, A.E., Walters, M.S., Shaykhiev, R., and Crystal, R.G. (2015). Cilia dysfunction in lung disease. Ann. Rev. Physiol. 77, 379-406. </p>
<p>Wanner, A., Salathe, M., and O'Riordan, T.G. (1996). Mucociliary clearance in the airways. Am. J. Respir. Crit. Care Med. 154, 1868-1902. </p>
<p>Yager, J., Chen, T.-M., and Dulfano, M.J. (1978). Measurement of frequency of ciliary beats of human respiratory epithelium. Chest 73, 627-633.</p>
<p>Yeates, D.B., Katwala, S.P., Daugird, J., Daza, A.V., and Wong, L.B. (1997). Excitatory and inhibitory neural regulation of tracheal ciliary beat frequency (CBF) activated by ammonia vapour and SO2. Ann. Occup. Hyg. 41, 736-744.</p>
2021-07-19T10:24:022023-03-24T08:17:45932f4980-6e58-40a1-bffc-0c8fc0c177e30748dafe-40c9-413d-b87b-5fb1cf4d8120<p>It is very well known that patients suffering from motile ciliopathies, such as primary ciliary dyskinesia, have impaired or absent MCC and lower lung function (reduced FEV1 and FVC) compared to their healthy counterparts (Halbeisen et al., 2018; Marthin et al., 2010; Wallmeier et al., 2020). In cystic fibrosis patients, decreased MCC (due to reduced airway hydration and changes in mucus chemical and viscoelastic properties) causes mucus build-up leading to mucus plugging in the airways and consequently to decreased lung function over time (Kerem et al., 2014; Mossberg et al., 1978; Regnis et al., 1994; Robinson and Bye, 2002; Szczesniak et al., 2017; Wanner et al., 1996). Mucus plugging due to decreased MCC is also considered a major cause of airway obstruction and airflow limitation in COPD patients (Dunican et al., 2021; Okajima et al., 2020) and asthmatics (Kuyper et al., 2003; Maxwell, 1985).</p>
<p>Changes in MCC rate are typically paralleled by effects on lung function in several studies where both endpoints have been assessed. In patients with primary ciliary dyskinesia, absence of cilia motion prevents normal MCC and consequently, lung function is reduced (Denizoglu Kulli et al., 2020). In cystic fibrosis patients, the ASL is depleted resulting in impaired MCC (Boucher, 2004). Although the known CFTR genotypes can result in a variety of phenotypes (Derichs, 2013), clinical data indicate that some specific gene defects, such as the p.Phe508del variant, are more frequently associated with decreased lung function indices (e.g. FEV1 % predicted, FVC % predicted, FEF25-75) (Kerem et al., 1990; Johansen et al., 1991; Schaedel et al., 2002). Both cigarette smoking and occupational exposure to biomass fumes led to slower MCC and reduced FEV1 % predicted and FEV1/FVC (Ferreira et al., 2018). Nasomucociliary clearance was slower in COPD smokers compared to former smokers with COPD or to nonsmokers (Ito et al., 2015). Allergen challenge in asthma patients resulted in both reduced MCC and FEV1, which could be reversed by inhalation of hypertonic saline solution (Alexis et al., 2017). In cystic fibrosis patients, treatment with mucolytic agents (Laube et al., 1996; McCoy et al., 1996; Quan et al., 2001; Elkins et al., 2006; Amin et al., 2011; Donaldson et al., 2018) or a CFTR potentiator (Rowe et al., 2014) improved both MCC and lung function (FEV1, FVC and FEF25-75).</p>
<p>Lung function is known to decrease with age, and several studies showed that mucus transport rates also decrease in older compared to younger individuals (Goodman et al., 1978; Uzeloto et al., 2021). Impaired MCC is also seen in chronic smokers, even prior to a clinically significant drop in lung function and the detection of small airway disease (Clunes et al., 2012a; Goodman et al., 1978; Lourenço et al., 1971; Uzeloto et al., 2021; Vastag et al., 1986), and in patients with obstructive lung disease and hence, poor lung function (Cruz et al., 1974; Vastag et al., 1986). Adult asthmatics also displayed decreased mucus transport rates/velocities in addition to decreased lung function (Ahmed et al., 1981; Bateman et al., 1983; Foster et al., 1982; Mezey et al., 1978).<br />
In patients with primary ciliary dyskinesia, absence of cilia motion prevents normal MCC and consequently, lung function is reduced (Denizoglu Kulli et al., 2020). In cystic fibrosis patients, the ASL is depleted resulting in impaired MCC (Boucher, 2004a). Although the known CFTR genotypes can result in a variety of phenotypes (Derichs, 2013), clinical data indicate that some specific gene defects, such as the p.Phe508del variant, are more frequently associated with decreased lung function indices (e.g. FEV1 % predicted, FVC % predicted, FEF25-75) (Johansen et al., 1991; Kerem et al., 1990; Schaedel et al., 2002). Unsurprisingly, results from studies with pharmacological agents aimed at restoring CFTR function do not only indicate enhanced MCC but also support improvements in lung function (Bennett et al., 2018; Donaldson et al., 2018; Rowe S. M. et al., 2014a). While the available data link these two KEs, causal evidence is not always available, and some inference is present. Therefore, we judge the biological plausibility of this KER as moderate.</p>
<p>Occupational exposure to biomass combustion products resulted in slower MCC and reduced FEV1 % predicted and FEV1/FVC (Ferreira et al., 2018).</p>
<p>Compared to healthy controls, current smokers without airway obstruction and current smokers with COPD exhibited longer saccharin transit times, indicative of impaired MCC, and lower FEV1 % predicted and FEV1/FVC (Uzeloto et al., 2021). Similarly, nasomucociliary clearance was slower in COPD smokers compared to former smokers with COPD or to nonsmokers (Ito et al., 2015). Additionally, mucus plug density—assessed by CT imaging—and mucoid (rather than watery) consistency were inversely related to FEF25–75% and associated with increased RV/TLV (Kesimer et al., 2018).</p>
<p>Asthma patients responded to allergen challenge with a reduction in both MCC and FEV1 (Bennett et al., 2011; Mezey et al., 1978), which could be rescued by inhalation with hypertonic saline solution (Alexis et al., 2017).</p>
<p>Multiple studies interrogating the effect of mucolytic agents such as hypertonic saline solution or recombinant DNase on mucus transport rates or mucus clearance in patients with cystic fibrosis report improvements in both, mucus transport velocities or rates and lung function indices, including FEV1, FVC and FEF25-75 (Amin et al., 2011; Donaldson et al., 2018; Elkins et al., 2006; Laube et al., 1996; McCoy et al., 1996; Quan et al., 2001).</p>
<p>Both MCC and lung function (FEV1, FVC and FEF25-75) improved in cystic fibrosis patients treated with ivacaftor, a CFTR potentiator that increases the channel open probability (Rowe et al., 2014b).</p>
<p>Some studies with mucolytics such as N-acetylcysteine, bromhexine, theophylline/ambroxol or sorebrol demonstrated improved MCC was connected with small improvements in lung function (FEV1, FVC and FEV1/FVC) in patients with chronic bronchitis (Aylward et al., 1980; Castigiioni and Gramolini, 1986; Thomson et al., 1974; Würtemberger et al., 1988).<br />
</p>
<p>Genetic defects leading to motile ciliopathies or defects in CFTR function are linked to impaired MCC. However, because of the genetic variety, not every defect, for example in the CFTR gene, also expresses an overt pulmonary phenotype. Other factors, such as low-level chronic inflammation may drive lung pathology by pathways independent of MCC. This might also explain the absence of differences in MCC between healthy smokers and smokers with COPD (Fleming et al., 2019).<br />
Not all studies looking to elucidate the effect of mucolytics on MCC report an improvement of lung function, even though mucus transport rates or tracheobronchial clearance significantly improve. These studies include, for example, some on the effects of hypertonic saline solution, NAC, ambroxol and 2-mercapto-ethane sulphonate (Clarke et al., 1979; Ericsson et al., 1987; Millar et al., 1985; Robinson et al., 1997; Würtemberger et al., 1988). This could be, at least in part, related to the fact that a sudden drop in lung function served as an indicator of patient distress in these studies, and interventions were halted when they occurred to ensure patient safety (Robinson et al., 1996). Another reason could be related to the mechanisms underlying mucus solubilization that may be completely independent of lung function.<br />
MCC is only one means by which mucus can be cleared from the lungs. Another one is cough clearance, and it is highly dependent on the properties of the ASL, in particular the ASL height (Knowles and Boucher, 2002).</p>
<p>The available data, though not causally linking decreases in MCC with decreased lung function, provide a good insight into the importance of the physiological role of MCC in maintaining normal lung function. In at least some studies, impairment of MCC correlated with the drop in FEV1 or FEF25-75. Although clinically valuable benefits can be seen in studies with pharmacological agents such as mucolytics and CFTR modifying drugs, they do not cover a wide range of dose responses nor are they supportive of the KER causality. Therefore, we judge our quantitative understanding as moderate.</p>
HighMixedHighAll life stagesHigh<p>The evidences for this KER come from and therefore apply to humans.</p>
<p>Ahmed, T., Greenblatt, D.W., Birch, S., Marchette, B., and Wanner, A. (1981). Abnormal mucociliary transport in allergic patients with antigen-induced bronchospasm: role of slow reacting substance of anaphylaxis. Am. Rev. Respir. Dis. 124, 110-114.</p>
<p>Alexis, N.E., Bennett, W., and Peden, D.B. (2017). Safety and benefits of inhaled hypertonic saline following airway challenges with endotoxin and allergen in asthmatics. J. Asthma 54, 957-960.</p>
<p>Amin, R., Subbarao, P., Lou, W., Jabar, A., Balkovec, S., Jensen, R., et al. (2011). The effect of dornase alfa on ventilation inhomogeneity in patients with cystic fibrosis. Eur. Respir. J. 37, 806-812.</p>
<p>Aylward, M., Maddock, J., and Dewland, P. (1980). Clinical evaluation of acetylcysteine in the treatment of patients with chronic obstructive bronchitis: a balanced double-blind trial with placebo control. Eur. J. Respir. Dis. Suppl. 111, 81-89.</p>
<p>Bateman, J., Pavia, D., Sheahan, N., Agnew, J., and Clarke, S. (1983). Impaired tracheobronchial clearance in patients with mild stable asthma. Thorax 38, 463-467.</p>
<p>Bennett, W.D., Zeman, K.L., Laube, B.L., Wu, J., Sharpless, G., Mogayzel, P.J., Jr., et al. (2018). Homogeneity of Aerosol Deposition and Mucociliary Clearance are Improved Following Ivacaftor Treatment in Cystic Fibrosis. J. Aerosol Med. Pulm. Drug Delivery 31, 204-211. </p>
<p>Bennett, W.D., Almond, M.A., Zeman, K.L., Johnson, J.G., and Donohue, J.F. (2006). Effect of salmeterol on mucociliary and cough clearance in chronic bronchitis. Pulmon. Pharmacol. Therap. 19, 96-100.</p>
<p>Bennett, W.D., Herbst, M., Alexis, N.E., Zeman, K.L., Wu, J., Hernandez, M.L., et al. (2011). Effect of inhaled dust mite allergen on regional particle deposition and mucociliary clearance in allergic asthmatics. Clin. Exp. Allergy 41, 1719-1728. </p>
<p>Boucher, R. (2004). New concepts of the pathogenesis of cystic fibrosis lung disease. Eur. Respir. J. 23, 146-158.</p>
<p>Button, B., Goodell, H.P., Atieh, E., Chen, Y.-C., Williams, R., Shenoy, S., et al. (2018). Roles of mucus adhesion and cohesion in cough clearance. Proc. Natl. Acad. Sci. U.S.A. 115, 12501-12506. </p>
<p>Clunes, L.A., Davies, C.M., Coakley, R.D., Aleksandrov, A.A., Henderson, A.G., Zeman, K.L., et al. (2012). Cigarette smoke exposure induces CFTR internalization and insolubility, leading to airway surface liquid dehydration. FASEB J. 26, 533-545. </p>
<p>Cruz, R.S., Landa, J., Hirsch, J., and Sackner, M.A. (1974). Tracheal mucous velocity in normal man and patients with obstructive lung disease; effects of terbutaline. Am. Rev. Respir. Dis. 109, 458-463.</p>
<p>Denizoglu Kulli, H., Gurses, H.N., Zeren, M., Ucgun, H., and Cakir, E. (2020). Do pulmonary and extrapulmonary features differ among cystic fibrosis, primary ciliary dyskinesia, and healthy children? Pediatr. Pulmonol. 55, 3067-3073. </p>
<p>Derichs, N. (2013). Targeting a genetic defect: cystic fibrosis transmembrane conductance regulator modulators in cystic fibrosis. Eur. Respir. J. 22, 58-65.</p>
<p>Donaldson, S.H., Bennett, W.D., Zeman, K.L., Knowles, M.R., Tarran, R., and Boucher, R.C. (2006). Mucus Clearance and Lung Function in Cystic Fibrosis with Hypertonic Saline. N. Engl. J. Med. 354, 241-250. </p>
<p>Donaldson, S.H., Laube, B.L., Corcoran, T.E., Bhambhvani, P., Zeman, K., Ceppe, A., et al. (2018). Effect of ivacaftor on mucociliary clearance and clinical outcomes in cystic fibrosis patients with G551D-CFTR. JCI Insight 3, e122695. </p>
<p>Dueholm, M., Nielsen, C., Thorshauge, H., Evald, T., Hansen, N.-C., Madsen, H., et al. (1992). N-acetylcysteine by metered dose inhaler in thetreatment of chronic bronchitis: a multi-centre study. Respir. Med. 86, 89-92.</p>
<p>Dunican, E.M., Elicker, B.M., Henry, T., Gierada, D.S., Schiebler, M.L., Anderson, W., et al. (2021). Mucus plugs and emphysema in the pathophysiology of airflow obstruction and hypoxemia in smokers. Am. J. Respir. Crit. Care Med. 203, 957-968.</p>
<p>Elkins, M.R., Robinson, M., Rose, B.R., Harbour, C., Moriarty, C.P., Marks, G.B., et al. (2006). A Controlled Trial of Long-Term Inhaled Hypertonic Saline in Patients with Cystic Fibrosis. N. Engl. J. Med. 354, 229-240. </p>
<p>Ferreira, A.D., Ramos, E.M.C., Trevisan, I.B., Leite, M.R., Proença, M., de Carvalho-Junior, L.C.S., et al. (2018). Função pulmonar e depuração mucociliar nasal de cortadores de cana-de-açúcar brasileiros expostos à queima de biomassa. Rev. Bras. Saúde Ocup. 43,e6.</p>
<p>Foster, W., Langenback, E., and Bergofsky, E. (1982). "Lung mucociliary function in man: interdependence of bronchial and tracheal mucus transport velocities with lung clearance in bronchial asthma and healthy subjects," in Inhaled Particles V. Elsevier), 227-244.</p>
<p>Goodman, R., Yergin, B., Landa, J., Golinvaux, M., and Sackner, M. (1978). Relationship of smoking history and pulmonary function tests to tracheal mucous velocity in nonsmokers, young smokers, ex-smokers, and patients with chronic bronchitis. Am. Rev. Respir. Dis. 117, 205-214.</p>
<p>Halbeisen, F.S., Goutaki, M., Spycher, B.D., Amirav, I., Behan, L., Boon, M., et al. (2018). Lung function in patients with primary ciliary dyskinesia: an iPCD Cohort study. Eur. Respir. J. 52, 1801040.</p>
<p>Hasani, A., Toms, N., O'Connor, J., Dilworth, J., and Agnew, J. (2003). Effect of salmeterol xinafoate on lung mucociliary clearance in patients with asthma. Respir. Med. 97, 667-671.</p>
<p>Ito, J.T., Ramos, D., Lima, F.F., Rodrigues, F.M., Gomes, P.R., Moreira, G.L., et al. (2015). Nasal Mucociliary Clearance in Subjects With COPD After Smoking Cessation. Respir. Care 60, 399-405. </p>
<p>Johansen, H.K., Nir, M., Koch, C., Schwartz, M., and Høiby, N. (1991). Severity of cystic fibrosis in patients homozygous and heterozygous for ΔF508 mutation. Lancet 337, 631-634.</p>
<p>Kerem, E., Corey, M., Kerem, B.-s., Rommens, J., Markiewicz, D., Levison, H., et al. (1990). The relation between genotype and phenotype in cystic fibrosis—analysis of the most common mutation (ΔF508). N. Engl. J. Med. 323, 1517-1522.</p>
<p>Kerem, E., Viviani, L., Zolin, A., MacNeill, S., Hatziagorou, E., Ellemunter, H., et al. (2014). Factors associated with FEV1 decline in cystic fibrosis: analysis of the ECFS Patient Registry. Eur. Respir. J. 43, 125-133. </p>
<p>Kesimer, M., Smith, B.M., Ceppe, A., Ford, A.A., Anderson, W.H., Barr, R.G., et al. (2018). Mucin concentrations and peripheral airway obstruction in chronic obstructive pulmonary disease. Am. J. Respir. Crit. Care Med. 198, 1453-1456.</p>
<p>King, M. (2006). Physiology of mucus clearance. Paediatr. Respir. Rev. 7 Suppl 1, S212-214. </p>
<p>Kuyper, L.M., Paré, P.D., Hogg, J.C., Lambert, R.K., Ionescu, D., Woods, R., et al. (2003). Characterization of airway plugging in fatal asthma. Am. J. Med. 115, 6-11.</p>
<p>Laube, B.L., Auci, R.M., Shields, D.E., Christiansen, D.H., Lucas, M.K., Fuchs, H.J., et al. (1996). Effect of rhDNase on airflow obstruction and mucociliary clearance in cystic fibrosis. Am. J. Respir. Crit. Care Med. 153, 752-760.</p>
<p>Lourenço, R.V., Klimek, M.F., and Borowski, C.J. (1971). Deposition and clearance of 2 μ particles in the tracheobronchial tree of normal subjects—smokers and nonsmokers. J. Clin. Invest. 50, 1411-1420.</p>
<p>Marthin, J.K., Petersen, N., Skovgaard, L.T., and Nielsen, K.G. (2010). Lung function in patients with primary ciliary dyskinesia: a cross-sectional and 3-decade longitudinal study. Am. J. Respir. Crit. Care Med. 181, 1262-1268.</p>
<p>Maxwell, G. (1985). The problem of mucus plugging in children with asthma. J. Asthma 22, 131-137.</p>
<p>McCoy, K., Hamilton, S., and Johnson, C. (1996). Effects of 12-Week Administration of Dornase Alfa in Patients with Advanced Cystic Fibrosis Lung Disease. Chest 110, 889-895. </p>
<p>Mezey, R.J., Cohn, M.A., Fernandez, R.J., Januszkiewicz, A.J., and Wanner, A. (1978). Mucociliary transport in allergic patients with antigen-induced bronchospasm. Am. Rev. Respir. Dis. 118, 677-684.</p>
<p>Mossberg, B., Afzelius, B., Eliasson, R., and Camner, P. (1978). On the pathogenesis of obstructive lung disease. A study on the immotile-cilia syndrome. Scand. J. Respir. Dis. 59, 55-65.</p>
<p>Okajima, Y., Come, C.E., Nardelli, P., Sonavane, S.K., Yen, A., Nath, H.P., et al. (2020). Luminal Plugging on Chest CT Scan: Association With Lung Function, Quality of Life, and COPD Clinical Phenotypes. Chest 158, 121-130. </p>
<p>Quan, J.M., Tiddens, H.A.W.M., Sy, J.P., McKenzie, S.G., Montgomery, M.D., Robinson, P.J., et al. (2001). A two-year randomized, placebo-controlled trial of dornase alfa in young patients with cystic fibrosis with mild lung function abnormalities. J. Pediatr. 139, 813-820. </p>
<p>Regnis, J., Robinson, M., Bailey, D., Cook, P., Hooper, P., Chan, H., et al. (1994). Mucociliary clearance in patients with cystic fibrosis and in normal subjects. Am. J. Respir. Crit. Care Med. 150, 66-71.</p>
<p>Robinson, M., and Bye, P.T.B. (2002). Mucociliary clearance in cystic fibrosis. Pediatr. Pulmonol. 33, 293-306. </p>
<p>Rowe, S.M., Heltshe, S.L., Gonska, T., Donaldson, S.H., Borowitz, D., Gelfond, D., et al. (2014). Clinical mechanism of the cystic fibrosis transmembrane conductance regulator potentiator ivacaftor in G551D-mediated cystic fibrosis. Am. J. Respir. Crit. Care Med. 190, 175-184.</p>
<p>Schaedel, C., de Monestrol, I., Hjelte, L., Johannesson, M., Kornfält, R., Lindblad, A., et al. (2002). Predictors of deterioration of lung function in cystic fibrosis. Pediatr. Pulmonol. 33, 483-491. </p>
<p>Szczesniak, R., Heltshe, S.L., Stanojevic, S., and Mayer-Hamblett, N. (2017). Use of FEV(1) in cystic fibrosis epidemiologic studies and clinical trials: A statistical perspective for the clinical researcher. J. Cyst. Fibros. 16, 318-326. </p>
<p>Thomson, M., Pavia, D., Gregg, I., and Stark, J. (1974). Bromhexine and mucociliary clearance in chronic bronchitis. Brit. J. Diseases Chest 68, 21-27.</p>
<p>Uzeloto, J.S., Ramos, D., Silva, B.S.d.A., Lima, M.B.P.d., Silva, R.N., Camillo, C.A., et al. (2021). Mucociliary Clearance of Different Respiratory Conditions: A Clinical Study. Int. Arch. Otorhinolaryngol. 25, e35-e40. </p>
<p>Vastag, E., Matthys, H., Zsamboki, G., Köhler, D., and Daikeler, G. (1986). Mucociliary clearance in smokers. Eur. J. Respir. Dis. 68, 107-113.</p>
<p>Wallmeier, J., Nielsen, K.G., Kuehni, C.E., Lucas, J.S., Leigh, M.W., Zariwala, M.A., et al. (2020). Motile ciliopathies. Nat. Rev. Dis. Prim. 6, 1-29.</p>
<p>Würtemberger, G., Michaelis, K., and Matthys, H. (1988). [Additive action of theophylline and ambroxol on bronchial clearance?]. Prax. Klin. Pneumol. 42 Suppl 1, 300-303.</p>
2021-07-19T10:24:312023-03-24T08:27:51Oxidative stress Leading to Decreased Lung Function Oxidative stress Leading to Decreased Lung Function<p>Karsta Luettich, Philip Morris Products S.A., Philip Morris International R&D, Neuchatel, Switzerland</p>
<p>Hasmik Yepiskoposyan, Philip Morris Products S.A., Philip Morris International R&D, Neuchatel, Switzerland</p>
<p>Monita Sharma, PETA Science Consortium International e.V., Stuttgart, Germany</p>
<p>Frazer Lowe, Broughton Nicotine Services, Earby, Lancashire, United Kingdom</p>
<p>Damien Breheny, British American Tobacco (Investments) Ltd., Group Research and Development, Southampton, United Kingdom<br />
</p>
Open for comment. Do not cite<p>We propose here an AOP that attempts to delineate how exposure to oxidative insults lead to decreased lung function (Luettich et al., 2021). This AOP evaluates one of the major processes known to be involved in regulating efficient mucociliary clearance (MCC)—ciliary function. MCC is a key aspect of the innate immune defense against airborne pathogens and inhaled chemicals and is governed by the concerted action of its functional components, the cilia and the airway surface liquid (ASL), which is composed of mucus and periciliary layers (Bustamante-Marin and Ostrowski, 2017). Disturbances in any of the processes regulating ciliary function can cause MCC dysfunction. Impaired MCC is linked to airway diseases such as chronic obstructive pulmonary disease (COPD) or asthma, both of which are characterized by decreased lung function and bear a significant risk of increased morbidity and mortality. Given the individual and public health burden of the consequences of lung function impairment, gaining a greater understanding of the underlying mechanisms is extremely important in the risk assessment of respiratory toxicants.</p>
<p>The KE proposed here are moderately to highly essential, and we judge the overall biological plausibility of this AOP as strong. The KER <em>Oxidative stress leading to decreased CBF</em> is supported by multiple studies across different species with ample empirical evidence reflecting both dose-response and time concordance. The KER <em>Decreased CBF leading to decreased MCC</em> lacks this expanse of empirical evidence, or the evidence does not fully support the causality between the KE even though the relationship is logical and plausible. Overall, our quantitative understanding of the AOP is moderate.</p>
<p>With a surface area of ~100 m<sup>2</sup> and ventilated by 10,000 to 20,000 liters of air per day (National Research Council, 1988; Frohlich et al., 2016), the lungs are a major barrier that protect the body from a host of external factors that enter the respiratory system and may cause lung pathologies. Mucociliary clearance (MCC) is a key aspect of the innate immune defense against airborne pathogens and inhaled particles and is governed by the concerted action of its functional components, the cilia and the airway surface liquid (ASL), which comprises mucus and the periciliary layer (Bustamante-Marin and Ostrowski, 2017). In healthy subjects, ≥10 mL airway secretions are continuously produced and transported daily by the mucociliary escalator. Disturbances in any of the processes regulating ASL volume, mucus production, mucus viscoelastic properties, or ciliary function can cause MCC dysfunction and are linked to airway diseases such as chronic obstructive pulmonary disease (COPD) or asthma, both of which bear a significant risk of increased morbidity and mortality. The mechanism by which exposure to inhaled toxicants might lead to mucus hypersecretion and thereby impact pulmonary function has already been mapped in AOP148 on decreased lung function. However, whether an exposure-related decline in lung function is solely related to excessive production of mucus is debatable, particularly in light of the close relationship between mucus, ciliary function, and efficient MCC. To date, no single event has been attributed to MCC impairment, and it is likely that events described in this AOP as well as in AOP148, AOP424, and AOP425 have to culminate to lead to decreased lung function.</p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Established regulatory guideline studies for inhalation toxicity focus on evident clinical signs of systemic toxicity, including death, or organ-specific toxicity following acute and (sub)chronic exposure respectively. In toxicological and safety pharmacological studies with airborne test items targeting the airways or the lungs as a whole, lung function is a relevant endpoint for the characterization of potential adverse events (OECD, 2018a; Hoymann, 2012). Hence, the AO “decreased lung function” is relevant for regulatory decision-making in the context of (sub)chronic exposure (OECD, 2018b; OECD, 2018c).</span></span></span></span></span></p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Regulatory relevance of the AO “decreased lung function” is evident when looking at the increased risk of diseases in humans following inhalation exposure, and because of its links to other comorbidities and mortality.</span></span></span></span></span></p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">To aid diagnosis and monitoring of fibrosis, current recommendations include both the recording of potential environmental and occupational exposures as well as an assessment of lung function (Baumgartner et al., 2000). The latter typically confirms decreased lung function as demonstrated by a loss of lung volume. As the disease progresses, dyspnea and lung function worsen, and the prognosis is directly linked to the decline in FVC (Meltzer and Noble, 2008). </span></span></span></span></span></p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="font-family:"Segoe UI",sans-serif"><span style="color:black">Chronic exposure to cigarette smoke and other combustion-derived particles results in the development of COPD. COPD is diagnosed on the basis of spirometry results as laid out in the ATS/ERS Task Force documents on the standardization of lung function tests and their interpretation (Pellegrino et al., 2005; Culver et al., 2017, Graham et al., 2019). Rapid rates of decline in the lung function parameter FEV1 are linked to higher risk of exacerbations, increased hospitalization and early death (Wise et al., 2006; Celli, 2010). Reduced FEV1 also poses a risk for serious cardiovascular events and mortality associated with cardiovascular disease (Sin et al., 2005; Lee et al., 2015).</span></span></span></span></span></p>
<p> </p>
adjacentHighHighadjacentModerateHighadjacentModerateModerate<p>The definition of essentiality implies that the modulation of upstream KEs impacts the downstream KEs in an expected fashion. If blocked or failing to occur, the KEs in the current AOP will not necessarily stop the progression to subsequent KEs. Due to the complex biology of motile cilia formation and function, ASL homeostasis, mucus properties and their concerted impact on MCC, the KEs and AO may be triggered because of alternative pathways or biological redundancies. However, when exacerbated, the KEs promote the occurrence of downstream events eventually leading to the AO. The causal pathway starting from the exposure to oxidants and leading to decreased lung function involves parallel routes with KEs, each of which is sufficient to cause the downstream KE to occur. Based on the evidence we judge the MIE (Oxidative Stress), KE1908 (Cilia Beat Frequency, Decreased), and KE1909 (Mucociliary Clearance, Decreased) as highly essential. </p>
Not SpecifiedMixedNot SpecifiedAll life stagesHigh<p>The experimental evidence to support the biological plausibility of the KERs from MIE to AO is moderate to strong overall for the AOP presented here, while there is a moderate concordance of dose-response relationships. In terms of essentiality, we have rated all of the KEs as either moderate or high.</p>
<p>AOPs such as this one can play a central role in risk assessment strategies for a wide variety of regulatory purposes by providing mechanistic support to an integrated approach to testing and assessment (IATA; (Clippinger et al., 2018)). IATAs are flexible frameworks that can be adapted to best address the regulatory question or purpose at hand. More specifically, this AOP can be applied to the risk assessment of inhaled toxicants, by enabling the development of testing strategies through the assembly of existing information and the generation of new data where they are currently lacking. Targeted approaches to fill data gaps can be developed using new approach methodologies (NAMs) informed by this AOP.</p>
<p>All KE proposed in this AOP occur and are measurable in several species, including frogs, mice, rats, guinea pigs, ferrets, cats, dogs, cows, monkeys, and humans. The majority of the supporting empirical evidence derives from studies in rodent and human systems, and experimental findings in animals appear to be highly translatable to humans.</p>
<p>Data regarding the applicability of KE to all life-stages from birth to adulthood are available for the MIE (Oxidative Stress), KE1908 (Cilia Beat Frequency, Decreased), KE1909 (Mucociliary Clearance, Decreased), and AO (Decreased Lung Function), and indicate that they apply to all life stages. It is also worth noting here that age-dependent decreases in CBF, MCC, and lung function have been demonstrated in several species (e.g., guinea pigs, mice, and humans) and reflect normal physiological aging processes (Bailey et al., 2014; Grubb et al., 2016; Ho et al., 2001; Joki and Saano, 1997; Paul et al., 2013; Sharma and Goodwin, 2006).</p>
<p>Gender-specific data relevant to the AOP network are not as widely available as species-specific data, and to our knowledge, the role of gender has not been systematically evaluated for all KE described here. Informative evidence on gender differences stems from patients with chronic pulmonary diseases, such as cystic fibrosis, asthma, COPD, and bronchiectasis, that are characterized by decreased lung function. Considering the importance of efficient MCC—brought about by the interactions of ciliary function, ASL homeostasis and mucus properties—for normal physiological function, we consider this AOP applicable to both genders.</p>
<p>The definition of essentiality implies that the modulation of upstream KEs impacts the downstream KEs in an expected fashion. If blocked or failing to occur, the KEs in the current AOP will not necessarily stop the progression to subsequent KEs. Due to the complex biology of motile cilia formation and function, ASL homeostasis, mucus properties and their concerted impact on MCC, the KEs and AO may be triggered because of alternative pathways or biological redundancies. However, when exacerbated, the KEs promote the occurrence of downstream events eventually leading to the AO. The causal pathway starting from the exposure to oxidants and leading to decreased lung function involves parallel routes with KEs, each of which is sufficient to cause the downstream KE to occur. Based on the evidence we judge the MIE (Oxidative Stress), KE1908 (Cilia Beat Frequency, Decreased), and KE1909 (Mucociliary Clearance, Decreased) as highly essential. </p>
<p>We judge the overall biological plausibility of this AOP as strong. The KER <em>Oxidative stress leading to decreased CBF</em> is supported by multiple studies across different species with ample empirical evidence reflecting both dose-response and time concordance. The KER <em>Decreased CBF leading to decreased MCC</em> lacks this expanse of empirical evidence, or the evidence does not fully support the causality between the KE even though the relationship is logical and plausible. </p>
<p>Overall, our quantitative understanding of the AOP network is moderate.</p>
<p>There is robust evidence that provides an insight into the KER <em>Oxidative stress leading to decreased cilia beat frequency</em> and <em>Decreased cilia beat frequency leading to decreased MCC</em>, and the dose response and temporal relationship between the two KE in question are well described and quantified for different stressors across different test systems. In some instances, we are less confident in our quantitative understanding. For example, dose response data as well as data supportive of the KE causality are limited for the KER <em>Decreased MCC leading to decreased lung function</em>. </p>
<p>Given the individual and public health burden of the consequences of lung function impairment, gaining a greater understanding of the underlying mechanisms is extremely important in the risk assessment of respiratory toxicants. An integrated assessment of substances with the potential to be inhaled, either intentionally or unintentionally, could incorporate inhalation exposure and dosimetry modelling to inform an in vitro approach with appropriate exposure techniques and cell systems to assess KEs in this AOP (EPA’s Office of Chemical Safety and Pollution Prevention, 2019). Standardization and robustness testing of assays against explicit performance criteria using suitable reference materials can greatly increase the level of confidence in their use for KE assessment (Petersen et al., 2021). Much of the empirical evidence that supports the KERs in the qualitative AOP described here was obtained from in vitro studies using well-established methodologies for biological endpoint assessment. Being chemical agnostic, this AOP can be applied to a variety of substances that share the AO. For example, impaired MCC and decreased lung function have a long-known relationship with smoking, but little is known about the consequences of long-term use of alternative inhaled nicotine delivery products such as electronic cigarettes and heated tobacco products. This AOP can form the basis of an assessment strategy to evaluate the effects of exposure to aerosol from these products based on the KEs identified here. </p>
ModerateModerateHighLowLow<p>Bailey, K.L., Bonasera, S.J., Wilderdyke, M., Hanisch, B.W., Pavlik, J.A., DeVasure, J., et al. (2014). Aging causes a slowing in ciliary beat frequency, mediated by PKCε. Am. J. Physiol. Lung Cell. Mol. Physiol. 306, L584-L589.</p>
<p>Bustamante-Marin, X.M., and Ostrowski, L.E. (2017a). Cilia and Mucociliary Clearance. Cold Spring Harb. Persp. Biol. 9, a028241. </p>
<p>Clippinger, A.J., Allen, D., Behrsing, H., BéruBé, K.A., Bolger, M.B., Casey, W., et al. (2018). Pathway-based predictive approaches for non-animal assessment of acute inhalation toxicity. Toxicol. In Vitro 52, 131-145.</p>
<p>EPA’s Office of Chemical Safety and Pollution Prevention (2019). "FIFRA Scientific Advisory Panel Meeting Minutes and Final Report No. 2019-01 Peer Review on Evaluation of a Proposed Approach to Refine the Inhalation Risk Assessment for Point of Contact Toxicity: A Case Study Using a New Approach Methodology (NAM) December 4 and 6, 2018 FIFRA Scientific Advisory Panel Meeting". U.S. Environmental Protection Agency).</p>
<p>Frohlich, E., Mercuri, A., Wu, S., and Salar-Behzadi, S. (2016). Measurements of Deposition, Lung Surface Area and Lung Fluid for Simulation of Inhaled Compounds. Front. Pharmacol. 7, 181. </p>
<p>Grubb, B.R., Livraghi-Butrico, A., Rogers, T.D., Yin, W., Button, B., and Ostrowski, L.E. (2016). Reduced mucociliary clearance in old mice is associated with a decrease in Muc5b mucin. Am. J. Physiol. Lung Cell. Mol. Physiol. 310, L860-L867.</p>
<p>Ho, J.C., Chan, K.N., Hu, W.H., Lam, W.K., Zheng, L., Tipoe, G.L., et al. (2001). The effect of aging on nasal mucociliary clearance, beat frequency, and ultrastructure of respiratory cilia. Am. J. Respir. Crit. Care Med. 163, 983-988.</p>
<p>Joki, S., and Saano, V. (1997). Influence of ageing on ciliary beat frequency and on ciliary response to leukotriene D4 in guinea-pig tracheal epithelium. Clin. Exp. Pharmacol. Physiol. 24, 166-169.</p>
<p>Luettich, K., Sharma, M., Yepiskoposyan, H., Breheny, D., and Lowe, F. J. (2021). An Adverse Outcome Pathway for Decreased Lung Function Focusing on Mechanisms of Impaired Mucociliary Clearance Following Inhalation Exposure. Frontiers in Toxicology, 55.</p>
<p>National Research Council (1988). Air Pollution, the Automobile, and Public Health. Washington, DC: The National Academies Press.</p>
<p>Paul, P., Johnson, P., Ramaswamy, P., Ramadoss, S., Geetha, B., and Subhashini, A. (2013). The effect of ageing on nasal mucociliary clearance in women: a pilot study. ISRN 2013, 598589.</p>
<p>Petersen, E.J., Sharma, M., Clippinger, A.J., Gordon, J., Katz, A., Laux, P., et al. (2021). Use of Cause-and-Effect Analysis to Optimize the Reliability of In Vitro Inhalation Toxicity Measurements Using an Air–Liquid Interface. Chem. Res. Toxicol. 34, 1370–1385.</p>
<p>Sharma, G., and Goodwin, J. (2006). Effect of aging on respiratory system physiology and immunology. Clin. Interv. Aging 1, 253-260. </p>
2021-07-19T09:29:142023-04-29T16:03:05