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Vuba aha, hateguwe uburyo bwa antimicrobial butagira imiti bushingiye kuri nanotehnologiya hakoreshejwe amazi ya nanostructures (EWNS). EWNS ifite hejuru cyane kandi ikungahaye ku bwoko bwa ogisijeni ikora (ROS) ishobora gukorana no kudakora mikorobe nyinshi, harimo na virusi zitera ibiryo. Hano herekanwa ko imitungo yabo mugihe cya synthesis ishobora gutunganywa neza kandi igashyirwa mubikorwa kugirango irusheho kunoza ubushobozi bwa antibacterial. Laboratoire ya EWNS yateguwe kugirango ihuze neza imiterere ya EWNS ihindura ibipimo bya synthesis. Ibiranga imitungo ya EWNS (kwishyuza, ingano, n'ibirimo ROS) byakozwe hakoreshejwe uburyo bugezweho bwo gusesengura. Byongeye kandi, mikorobe y'ibiribwa nka Escherichia coli, Salmonella enterica, Listeria innocua, Mycobacterium para fortitum, na Saccharomyces cerevisiae yatewe hejuru yinyanya zinzabibu kama kugirango isuzume ubushobozi bwa mikorobe. Ibisubizo byatanzwe hano byerekana ko imiterere ya EWNS ishobora gutunganywa neza mugihe cya synthesis, bikavamo kwiyongera kugaragara mubikorwa byo kudakora neza. By'umwihariko, hejuru yubuso bwiyongereyeho ibintu bine, kandi ROS yariyongereye. Igipimo cyo gukuraho mikorobe cyari gishingiye kuri mikorobe kandi kuva kuri 1.0 kugeza kuri 3.8 nyuma yiminota 45 yo guhura na dose ya aerosol ya 40.000 # / cm3 EWNS.
Kwanduza mikorobe nimpamvu nyamukuru itera indwara ziterwa nibiribwa ziterwa no gufata virusi cyangwa uburozi bwabo. Indwara ziterwa n’ibiribwa zigera kuri miliyoni 76 z’indwara, 325.000 mu bitaro, n’impfu 5.000 buri mwaka muri Amerika yonyine1. Byongeye kandi, Minisiteri y’ubuhinzi muri Amerika (USDA) ivuga ko kongera umusaruro w’ibicuruzwa bishya ari byo nyirabayazana wa 48 ku ijana by’indwara zose ziterwa n’ibiribwa bivugwa muri Amerika2. Ikiguzi cy’indwara n’urupfu ziterwa na virusi ziterwa n’ibiribwa muri Amerika ni kinini cyane, ugereranije n’ikigo gishinzwe kurwanya no gukumira indwara (CDC) kirenga miliyari 15.6 z’amadolari y’Amerika ku mwaka3.
Kugeza ubu, imiti4, imirasire5 hamwe n’ubushyuhe bwa mikorobe 6 kugira ngo umutekano w’ibiribwa ushyirwe mu bikorwa aho bigenzurwa cyane (CCP) mu rwego rw’umusaruro (ubusanzwe nyuma yo gusarura no / cyangwa mu gihe cyo gupakira) aho gukomeza gushyirwa mu bikorwa ku buryo umusaruro mushya ushobora kwanduzwa 7. Kwifashisha imiti igabanya ubukana bw’ibiribwa no kwangirika kw’ibihingwa kandi bikagira ubushobozi bwo gukoreshwa ku meza. Ingaruka nke nigiciro.
Hashyizweho uburyo bwa nanotehnologiya bushingiye ku miti idafite imiti igabanya ubukana bwa virusi kugira ngo idakora bagiteri hejuru y’ikirere no mu kirere ikoresheje amazi ya nanostructures (EWNS). Kuri synthesis ya EVNS, hakoreshejwe inzira ebyiri zibangikanye: electrospray na ionisation y'amazi (Ishusho 1a). EWNS mbere yerekanwe ko ifite urutonde rwihariye rwimiterere yumubiri nibinyabuzima8,9,10. EWNS ifite impuzandengo ya electron 10 kuri buri miterere hamwe nubunini bwa nanometero ingana na 25 nm (Ishusho 1b, c) 8,9,10. Byongeye kandi, electron spin resonance (ESR) yerekanye ko EWNS irimo ubwoko bwinshi bwubwoko bwa ogisijeni (ROS), cyane cyane hydroxyl (OH •) na radicals ya superoxide (O2-) 8. EWNS yagumye mu kirere igihe kirekire kandi ishobora kugongana na mikorobe zahagaritswe mu kirere kandi zikagaragara ku buso, zigatanga imitwaro ya ROS kandi bigatuma mikorobe idakora (Ishusho 1d). Ubu bushakashatsi bwakozwe mbere bwerekanye kandi ko EWNS ishobora gukorana no gukora bagiteri zitandukanye za garama-mbi na gram-positif zifite akamaro k’ubuzima rusange bw’abaturage, harimo na mycobacteria, hejuru no mu kirere8,9. Ikwirakwizwa rya electron microscopi ryerekanaga ko kudakora byatewe no guhagarika ingirabuzimafatizo. Byongeye kandi, ubushakashatsi bukabije bwo guhumeka bwerekanye ko ibipimo byinshi bya EWNS bidatera kwangirika kw ibihaha cyangwa gutwika8.
(a) Electrospray ibaho mugihe hashyizweho ingufu nyinshi hagati ya capillary irimo amazi na compte electrode. . (c) Imiterere yihariye ya EWNS. (d) EWNS igendanwa cyane kubera imiterere ya nanoscale kandi irashobora gukorana nindwara ziterwa na virusi.
Ubushobozi bwa porogaramu ya mikorobe ya EWNS yo kudakora mikorobe ikomoka ku biribwa hejuru y’ibiribwa bishya nabyo byagaragaye vuba aha. Byerekanwe kandi ko EWNS yishyurwa hejuru irashobora gukoreshwa hamwe numurima wamashanyarazi mugutanga intego. Icy'ingenzi cyane, ni ibisubizo byambere byatanze hafi 1.4 yo kugabanya ibikorwa byinyanya kama yibikorwa byibinyabuzima bitandukanye nka E. coli na Listeria byagaragaye muminota 90 nyuma yo guhura na EWNS ku gipimo cya 50.000 # / cm311. Byongeye kandi, ibizamini byo gusuzuma ibinyabuzima byabanje kwerekana ko nta ngaruka ya organoleptic ugereranije ninyanya yo kugenzura. Nubwo ibisubizo byambere byo kudakora byizeza umutekano wibiryo ndetse no kuri EWNS nkeya cyane ya 50.000 # / cc. reba, biragaragara ko ubushobozi buke bwo kudakora byagira akamaro kurushaho kugirango ugabanye ibyago byo kwandura no kwangirika.
Hano, tuzibanda kubushakashatsi bwacu mugutezimbere urubuga rwa EWNS kugirango duhuze neza ibipimo bya synthesis kandi tunoze imitekerereze ya fiziki ya EWNS kugirango twongere ubushobozi bwa antibacterial. By'umwihariko, optimizasiyo yibanze ku kongera amafaranga yubuso bwabo (kunoza imitangire igenewe) hamwe nibirimo ROS (kunoza imikorere idahwitse). Kuranga ibintu byiza bya fiziki-chimique (ingano, amafaranga hamwe nibirimo ROS) ukoresheje uburyo bugezweho bwo gusesengura no gukoresha mikorobe isanzwe y'ibiribwa nka E. coli, S. enterica, L. innocua, S. cerevisiae na M. parafortuitum.
EVNS yashizwemo icyarimwe electrospraying hamwe na ionisation y'amazi meza cyane (18 MΩ cm - 1). Amashanyarazi atomizer 12 asanzwe akoreshwa muguhindura atomike ya polymer na synthique polymer hamwe nuduce twa ceramic 13 na fibre 14 yubunini bugenzurwa.
Nkuko byasobanuwe mubitabo byabanjirije 8, 9, 10, 11, mubigeragezo bisanzwe, ingufu nyinshi zikoreshwa hagati ya capillary yicyuma na electrode ihagaze. Muri iki gikorwa, habaho ibintu bibiri bitandukanye: 1) electrospray na 2) ionisation yamazi. Umuriro w'amashanyarazi ukomeye hagati ya electrode zombi zitera umuriro mubi kwiyongera hejuru y'amazi yegeranye, bigatuma habaho imishwarara ya Taylor. Kubera iyo mpamvu, hashyizweho ibitonyanga byamazi byuzuye, bikomeza gucikamo uduce duto, nkurikije inyigisho ya Rayleigh16. Muri icyo gihe, umurima w'amashanyarazi ukomeye utera molekile zimwe na zimwe z'amazi gucamo ibice no gukuramo electron (ionisation), bityo bikabyara amoko menshi ya ogisijeni ikora (ROS) 17. Icyarimwe icyarimwe ROS18 yamapaki yashyizwe muri EWNS (Ishusho 1c).
Ku mutini. 2a yerekana sisitemu ya EWNS ya sisitemu yateye imbere kandi ikoreshwa muri synthesis ya EWNS muri ubu bushakashatsi. Amazi meza yabitswe mu icupa rifunze yagaburiwe akoresheje umuyoboro wa Teflon (diameter 2 mm imbere) kugeza urushinge rwa 30G rutagira umuyonga (capillary yicyuma). Nkuko bigaragara ku gishushanyo cya 2b, amazi atemba agenzurwa numuvuduko wumwuka uri mumacupa. Urushinge rwometse kuri konsole ya Teflon ishobora guhindurwa nintoki intera runaka na compte electrode. Counter electrode ni disiki ya aluminiyumu isukuye ifite umwobo hagati kugirango itangwe. Munsi ya compte electrode ni aluminiyumu yerekana icyitegererezo, ihujwe nubundi buryo bwo kugerageza hakoreshejwe icyambu (Ishusho 2b). Ibice byose bya sampler byashizwe mumashanyarazi kugirango birinde kwishyurwa bishobora gutesha agaciro uduce duto duto.
(a) Sisitemu Yamazi Yububiko bwa Nanostructure Sisitemu (EWNS). (b) Igice cyambukiranya sampler na electrospray igice cyerekana ibipimo byingenzi. (c) Ubushakashatsi bwakorewe bagiteri idakora.
Sisitemu yo kubyara EWNS yasobanuwe haruguru irashoboye guhindura ibipimo byingenzi bikora kugirango byoroherezwe neza imitungo ya EWNS. Hindura voltage ikoreshwa (V), intera iri hagati y'urushinge na compte electrode (L), n'amazi atemba (φ) unyuze muri capillary kugirango uhuze neza ibiranga EWNS. Ibimenyetso [V (kV), L (cm)] bikoreshwa mugusobanura ibintu bitandukanye. Hindura amazi kugirango ubone cone ihamye ya Taylor runaka (V, L]. Kugira ngo ubu bushakashatsi bugerweho, aperture ya compte electrode (D) yashyizwe kuri santimetero 0,5 (cm 1,29).
Bitewe na geometrie ntarengwa hamwe na asimmetrie, imbaraga z'umuriro w'amashanyarazi ntizishobora kubarwa uhereye kumahame ya mbere. Ahubwo, porogaramu ya QuickField ™ (Svendborg, Danemarke) 19 yakoreshejwe mu kubara umurima w'amashanyarazi. Umuriro w'amashanyarazi ntabwo ari umwe, bityo agaciro k'umuriro w'amashanyarazi hejuru ya capillary wakoreshejwe nk'agaciro kerekana ibintu bitandukanye.
Muri ubwo bushakashatsi, hasuzumwe ibice byinshi bya voltage nintera iri hagati yinshinge na compte electrode ya compteur hashingiwe ku miterere ya cone ya Taylor, ihame rya cone ihagaze neza, umusaruro wa EWNS, n’imyororokere. Ihuriro ritandukanye ryerekanwe kumeza yinyongera S1.
Ibisohoka muri sisitemu yo kubyara EWNS byahujwe mu buryo butaziguye na Scanning Mobility Particle Sizer (SMPS, moderi 3936, TSI, Shoreview, Minnesota) kugirango bapime ingano yibice kandi yakoreshejwe hamwe na electrometero ya Faraday (TSI, icyitegererezo 3068B, Shoreview, USA). MN) gupima uruzinduko rwa aerosol, nkuko byasobanuwe mubitabo byabanje9. Byombi SMPS na electrometero ya aerosol byapimwe ku gipimo cya 0.5 L / min (icyitegererezo cyuzuye 1 L / min). Ibice byibanze hamwe na aerosol fluxes byapimwe kuri s 120. Subiramo ibipimo inshuro 30. Amafaranga yose ya aerosol abarwa uhereye kubipimo bigezweho, kandi impuzandengo ya EWNS igereranijwe uhereye ku mubare wuzuye wa EWNS watoranijwe. Impuzandengo yikiguzi cya EWNS irashobora kubarwa ukoresheje Ikigereranyo (1):
aho IEl ni igipimo cyapimwe, NSMPS numubare wapimwe hamwe na SMPS, na φEl nigipimo cyo gutemba kuri electrometero.
Kubera ko ubushuhe bugereranije (RH) bugira ingaruka ku buso, ubushyuhe na (RH) bwagumishijwe kuri 21 ° C na 45%, mugihe cyubushakashatsi.
Microscopi yingufu za kirimbuzi (AFM), Ubuhunzi MFP-3D (Ubushakashatsi bwubuhunzi, Santa Barbara, CA) na probe ya AC260T (Olympus, Tokiyo, Ubuyapani) byakoreshejwe mugupima ingano nubuzima bwa EWNS. Igipimo cya scan ya AFM ni 1 Hz naho scan ni 5 µm × 5 µm hamwe numurongo wa 256. Amashusho yose yakorewe gahunda yo gutondekanya bwa mbere amashusho akoresheje software yubuhunzi (mask ifite intera ya 100 nm hamwe ninjoro ya 100 pm).
Kuraho icyerekezo cya sample hanyuma ushire hejuru ya mika intera ya cm 2,2 uvuye kuri compte electrode mugihe cyo hagati ya 120 s kugirango wirinde guhuriza hamwe ibice no gushiraho ibitonyanga bidasanzwe hejuru ya mika. EWNS yakoreshwaga mu buryo butaziguye kuri mika yaciwe (Ted Pella, Redding, CA). Ako kanya nyuma yo gusakara, hejuru ya mika yagaragaye hakoreshejwe AFM. Ubuso bwo guhuza impande ya mika yaciwe vuba idahinduwe yegereye 0 °, EWNS rero ikwirakwiza hejuru ya mika muburyo bugaragara20. Diameter (a) n'uburebure (h) by'ibitonyanga bikwirakwizwa byapimwe biturutse kuri topografiya ya AFM kandi bikoreshwa mukubara ubwinshi bwikwirakwizwa rya EWNS dukoresheje uburyo bwemewe mbere8. Dufashe ko ku kibaho EVNS ifite ingano imwe, diameter ingana irashobora kubarwa uhereye ku kuringaniza (2):
Dukurikije uburyo twatunganije mbere, umutego wa spin spin resonance (ESR) umutego wa spin wakoreshejwe kugirango hamenyekane ko hari abahuza ba radical bahuza igihe gito muri EWNS. Aerosole yanyujijwe mu gisubizo kirimo 235 mM DEPMPO (5- (diethoxyphosphoryl) -5-methyl-1-pyrroline-N-oxide) (Oxis International Inc, Portland, Oregon). Ibipimo byose bya EPR byakozwe hakoreshejwe Bruker EMX yerekanwe (Bruker Instruments Inc. Billerica, MA, USA) hamwe na selile igaragara. Porogaramu ya Acquisit (Bruker Instruments Inc. Billerica, MA, USA) yakoreshejwe mu gukusanya no gusesengura amakuru. Ibiranga ROS byakozwe gusa kumurongo wimikorere [-6.5 kV, cm 4.0]. Ibipimo bya EWNS byapimwe hifashishijwe SMPS nyuma yo kuzirikana igihombo cya EWNS mubitera.
Urwego rwa Ozone rwakurikiranwe hifashishijwe Monitor ya 205 Dual Beam Ozone ™ (2B Technologies, Boulder, Co) 8,9,10.
Kubintu byose bya EWNS, agaciro ko gupima nuburyo bwo gupima, kandi ikosa ryo gupimwa niryo gutandukana bisanzwe. T-ikizamini cyakozwe kugirango ugereranye agaciro ka EWNS yatezimbere hamwe nagaciro kangana na EWNS.
Igicapo 2c cerekana mbere kandi cyaranzwe na Electrostatic Precipitation Pass Binyuze muri Sisitemu (EPES) zishobora gukoreshwa muguhitamo EWNS11 hejuru. EPES ikoresha amafaranga ya EWNS ifatanije numurima ukomeye wamashanyarazi kugirango "werekeza" hejuru yintego. Ibisobanuro bya sisitemu ya EPES bitangwa mubitabo biherutse gukorwa na Pyrgiotakis et al.11. Rero, EPES igizwe nicyumba cya 3D cyacapwe nicyumba cya PVC gifite impera zifunze zirimo ibyuma bibiri bisa nicyuma (304 ibyuma bitagira umuyonga, indorerwamo isize) icyuma hagati ya cm 15.24 zitandukanye. Ikibaho cyahujwe n’isoko ryo hejuru ya voltage yo hanze (Bertran 205B-10R, Spellman, Hauppauge, NY), ikibaho cyo hasi cyahoraga ari cyiza kandi ikibaho cyo hejuru cyahoraga gishyizwe hasi (kireremba). Urukuta rwa chambre rutwikiriwe na aluminiyumu, iba ifite amashanyarazi kugirango birinde gutakaza ibice. Icyumba gifite urugi rufunze imbere yipakurura ituma ibizamini bishyirwa kumurongo wa plastiki, ukabikura hejuru yicyuma cyo hasi kugirango wirinde kwivanga kwinshi.
Uburyo bwiza bwo kubitsa EWNS muri EPES bwabazwe ukurikije protocole yakozwe mbere irambuye mubisobanuro by'inyongera S111.
Nkicyumba cyo kugenzura, icya kabiri gitembera mucyumba cya silindrike gihujwe murukurikirane na sisitemu ya EPES ukoresheje akayunguruzo ka HEPA hagati kugirango ukureho EWNS. Nkuko bigaragara ku gishushanyo. 2c, aerosol ya EWNS yavomwe mubyumba bibiri bihujwe murukurikirane. Akayunguruzo hagati yicyumba cyo kugenzura na EPES gikuraho EWNS isigaye bivamo ubushyuhe bumwe (T), ubushuhe bugereranije (RH) nurwego rwa ozone.
Ibinyabuzima byingenzi byangiza ibiribwa byagaragaye ko byanduza umusaruro mushya nka Escherichia coli (ATCC # 27325), ikimenyetso cyerekana fecal, Salmonella enterica (ATCC # 53647), indwara yanduza ibiryo, Listeria innocua (ATCC # 33090), ubundi buryo bwo gutera indwara ya Listeria monocytogene. , Saccharomyces cerevisiae (ATCC # 4098) nk'uburyo bwo kwangiza umusemburo, na Mycobacterium parafortuitous (ATCC # 19686) nka bagiteri nzima irwanya indwara yaguzwe muri ATCC (Manassas, Virginia).
Gura kugura agasanduku k'inyanya z'imizabibu kama ku isoko ryaho hanyuma ugakonjesha kuri 4 ° C kugeza ukoresheje (kugeza ku minsi 3). Hitamo inyanya kugirango ugerageze nubunini bumwe, hafi 1/2 cy'uburebure.
Porotokole yo kubara, gukingirwa, kwerekana no kubara abakoloni byasobanuwe mu bitabo byabanjirije iki kandi bisobanurwa mu buryo burambuye mu makuru y’inyongera 11. Imikorere ya EWNS yasuzumwe no kwerekana inyanya zatewe kugeza 40.000 # / cm3 mu minota 45. Muri make, mugihe t = 0 min, inyanya eshatu zakoreshejwe mugusuzuma mikorobe ikiriho. Inyanya eshatu zashyizwe muri EPES kandi zerekanwa na EWNS kuri 40.000 # / cc (EWNS inyanya zashyizwe ahagaragara) izindi eshatu zishyirwa mubyumba bigenzura (inyanya zo kugenzura). Nta tsinda ry'inyanya ryakorewe izindi nyongera. EWNS yerekanwe inyanya nubugenzuzi byavanyweho nyuma yiminota 45 kugirango isuzume ingaruka za EWNS.
Buri igeragezwa ryakozwe muri bitatu. Isesengura ryamakuru ryakozwe ukurikije protocole yasobanuwe mu makuru yinyongera.
E. coli, Enterobacter, na L. innocua za bagiteri zanduye zanduye EWNS (iminota 45, EWNS ya aerosol yibanze 40.000 # / cm3) kandi zidashyizwe ahagaragara zarasuzumwe kugirango zisuzume uburyo bwo kudakora. Imvura yagizwe amasaha 2 mubushyuhe bwicyumba muri 0.1 M sodium cacodylate yumuti (pH 7.4) hamwe na glutaraldehyde ya 2.5%, paraformaldehyde 1,25% na acide picricike 0,03%. Nyuma yo gukaraba, bakosowe hamwe na 1% osmium tetroxide (OsO4) /1.5% potassium ferrocyanide (KFeCN6) kumasaha 2 h, bogejwe inshuro 3 namazi hanyuma bashiramo muri acetate ya uranyl 1% kuri 1 h, hanyuma bogejwe kabiri namazi. Umwuma ukurikiraho iminota 10 buri umwe muri 50%, 70%, 90%, 100% inzoga. Ingero zahise zishyirwa muri oxyde ya propylene kumasaha 1 hanyuma yinjizwa hamwe na 1: 1 ivanze na oxyde ya propylene na TAAP Epon (Marivac Canada Inc St. Laurent, CA). Ibyitegererezo byashyizwe muri TAAB Epon hanyuma bigashyirwa kuri 60 ° C mumasaha 48. Ibisigarira bya granular byakize byaciwe kandi bigaragazwa na TEM ukoresheje JEOL 1200EX (JEOL, Tokiyo, Ubuyapani), microscope isanzwe ikwirakwizwa ifite kamera ya AMT 2k CCD (Advanced Microscopy Technique, Corp., Woburn, MA, USA).
Ubushakashatsi bwose bwakozwe muri bitatu. Kuri buri mwanya, gukaraba kwa bagiteri byashyizwemo inshuro eshatu, bikavamo amanota icyenda yose hamwe kuri buri ngingo, impuzandengo yazo zikoreshwa nka bagiteri yibanda kuri kiriya kinyabuzima runaka. Gutandukana bisanzwe byakoreshejwe nkikosa ryo gupima. Ingingo zose zibara.
Logarithm yo kugabanuka kwinshi kwa bagiteri ugereranije na t = 0 min yabazwe hakoreshejwe formula ikurikira:
aho C0 yibanda kuri bacteri murugero rwo kugenzura mugihe 0 (nukuvuga nyuma yubuso bwumutse ariko mbere yo gushyirwa mubyumba) na Cn ni kwibumbira kwa bagiteri hejuru yiminota nyuma yiminota.
Kugirango habeho kwangirika kwa bagiteri mu gihe cyiminota 45, Log-Reduction nayo yabazwe ugereranije no kugenzura muminota 45 kuburyo bukurikira:
Aho Cn yibanda kuri bagiteri muburyo bwo kugenzura mugihe n na Cn-Igenzura ni ihuriro rya bagiteri igenzura mugihe n. Amakuru yatanzwe nkigabanuka ryibiti ugereranije no kugenzura (nta EWNS igaragara).
Muri ubwo bushakashatsi, hasuzumwe ibice byinshi bya voltage nintera iri hagati yinshinge na compte electrode ya compteur hashingiwe ku miterere ya cone ya Taylor, ihame rya cone ihagaze neza, umusaruro wa EWNS, n’imyororokere. Ihuriro ritandukanye ryerekanwe kumeza yinyongera S1. Imanza ebyiri zatoranijwe kubushakashatsi bwuzuye bwerekana ibintu bihamye kandi byororoka (Taylor cone, umusaruro wa EWNS, hamwe nigihe gihamye mugihe). Ku mutini. 3 yerekana ibisubizo kumafaranga, ingano n'ibirimo muri ROS kubibazo bibiri. Ibisubizo nabyo byavunaguye mu mbonerahamwe ya 1. Kubisobanuro, Igishushanyo cya 3 na Imbonerahamwe ya 1 birimo imitungo ya synthesize mbere idashimishije EWNS8, 9, 10, 11 (baseline-EWNS). Ibarurishamibare risobanura imibare ukoresheje imirongo ibiri t-igeragezwa ryongeye gutangazwa mu mbonerahamwe ya S2. Mubyongeyeho, amakuru yinyongera arimo ubushakashatsi ku ngaruka za compte electrode yerekana umwobo wa diameter (D) nintera iri hagati ya electrode yubutaka nisonga rya inshinge (L) (Ishusho yinyongera S2 na S3).
(a - c) Ikwirakwizwa rya AFM. (d - f) Ibiciro byo hejuru biranga. (g) Kuranga ROS na ESR.
Ni ngombwa kandi kumenya ko kubintu byose byavuzwe haruguru, imigezi yapimwe ionisiyoneri yapimwe yari hagati ya 2-6 µA, na voltage yari hagati ya -3.8 kugeza kuri -6.5 kV, bikavamo gukoresha amashanyarazi kuriyi EWNS imwe itarenza mW 50. . Module. Nubwo EWNS yashizwemo munsi yumuvuduko mwinshi, urugero rwa ozone rwari ruto cyane, ntirwigeze rurenga 60 ppb.
Ishusho yinyongera S4 yerekana imashanyarazi yigana amashanyarazi ya [-6.5 kV, cm 4.0] na [-3.8 kV, cm 0,5]. Imirima ukurikije ssenariyo [-6.5 kV, cm 4.0] na [-3.8 kV, 0,5 cm] ibarwa nka 2 × 105 V / m na 4.7 × 105 V / m. Ibi nibiteganijwe, kubera ko igipimo cya voltage nintera kiri hejuru cyane murwego rwa kabiri.
Ku mutini. 3a, b yerekana diameter ya EWNS yapimwe na AFM8. Impuzandengo ya EWNS ya diametre kuri [-6.5 kV, cm 4.0] na [-3.8 kV, cm 0,5] zabazwe nka 27 nm na 19 nm. Gutandukana kwa geometrike kugabanwa kubibazo [-6.5 kV, cm 4.0] na [-3.8 kV, cm 0,5] ni 1.41 na 1.45, byerekana ko ingano yagabanijwe. Ubunini buringaniye hamwe na geometrike itandukanijwe yegeranye cyane na baseline-EWNS, kuba 25 nm na 1.41. Ku mutini. 3c yerekana ingano yikwirakwizwa ryibanze EWNS yapimwe ukoresheje uburyo bumwe mubihe bimwe.
Ku mutini. 3d, e yerekana ibisubizo biranga amafaranga. Ibyatanzwe ni ibipimo bipima 30 icyarimwe icyarimwe cyo kwibanda (# / cm3) nubu (I). Isesengura ryerekana ko impuzandengo yikigereranyo kuri EWNS ari 22 ± 6 e- na 44 ± 6 e- kuri [-6.5 kV, cm 4.0] na [-3.8 kV, cm 0,5]. Ugereranije na Baseline-EWNS (10 ± 2 e-), ubwinshi bwubuso bwabo buri hejuru cyane, bwikubye kabiri ibya [-6.5 kV, cm 4.0] hamwe ninshuro enye za [-3 .8 kV, cm 0,5]. 3f yerekana amakuru yibanze ya EWNS.
Uhereye ku ikarita ya EWNS yerekana ikarita (Ishusho yinyongera S5 na S6), urashobora kubona ko [-6.5 kV, cm 4.0 cm] ifite umubare munini cyane wibice ugereranije na [-3.8 kV, cm 0,5]. Twabibutsa kandi ko umubare wa EWNS wakurikiranwe mu masaha agera kuri 4 (Ishusho yinyongera S5 na S6), aho ituze rya EWNS ryerekanaga urwego rumwe rwimibare yibice byombi.
Igishushanyo cya 3g cyerekana ecran ya EPR nyuma yo kugenzura (inyuma) gukuramo EWNS nziza kuri [-6.5 kV, cm 4.0]. Ikirangantego cya ROS nacyo cyagereranijwe na EWNS ibanze mu mpapuro zasohotse mbere. Umubare wabazwe wa EWNS witwara hamwe numutego wizunguruka ni 7.5 × 104 EWNS / s, bisa na Baseline-EWNS8 yatangajwe mbere. EPR yerekanwe neza ko hariho ubwoko bubiri bwa ROS, aho O2- yiganjemo, mugihe OH • yari ihari muke. Byongeye kandi, kugereranya mu buryo butaziguye ubukana bwa mpinga byerekanaga ko EWNS yatunganijwe neza yari ifite ibintu byinshi bya ROS ugereranije na EWNS y'ibanze.
Ku mutini. 4 yerekana uburyo bwo kubika EWNS muri EPES. Amakuru nayo yakusanyirijwe mu mbonerahamwe ya I kandi ugereranije namakuru yambere ya EWNS. Kubibazo byombi bya EUNS, kubitsa byari hafi 100% no kuri voltage ntoya ya 3.0 kV. Mubisanzwe, 3.0 kV irahagije kugirango ugere kubutaka 100% utitaye kumahinduka yubuso. Mubihe bimwe, imikorere yo kubitsa ya Baseline-EWNS yari 56% gusa kubera amafaranga make (impuzandengo ya electron 10 kuri EWNS).
Igishushanyo cya 5 nimbonerahamwe ya 2 byerekana muri make urugero rwo kudakora mikorobe zatewe hejuru yinyanya nyuma yo guhura na 40.000 # / cm3 EWNS muminota 45 mugihe cyiza [-6.5 kV, cm 4.0]. E. coli yatewe na L. innocua yerekanye igabanuka rikomeye rya 3.8 nyuma yiminota 45 yerekanwe. Mubihe bimwe, S. enterica yerekanye igabanuka ryo hasi ryibiti 2.2, mugihe S. cerevisiae na M. parafortuitum berekanye igabanuka rya 1.0.
Micrografi ya elegitoronike (Igishusho 6) yerekana impinduka zumubiri zatewe na EWNS muri E. coli, Salmonella enterica, na L. innocua selile zitera kudakora. Indwara ya bagiteri yerekanaga utugingo ngengabuzima tudakomeye, mu gihe bagiteri zagaragaye zangiritse hanze.
Electron microscopique yerekana amashusho hamwe na bagiteri zerekanwe byangiritse.
Ibyatanzwe kumiterere ya fiziki ya chimique ya EWNS yateranijwe hamwe yerekana ko imitungo ya EWNS (hejuru yubutaka hamwe nibirimo ROS) yatejwe imbere cyane ugereranije namakuru yibanze ya EWNS yatangajwe mbere 8,9,10,11. Kurundi ruhande, ubunini bwabo bwagumye murwego rwa nanometero, bisa cyane nibisubizo byatangajwe mbere, bibemerera kuguma mu kirere igihe kirekire. Polydispersity yagaragaye irashobora gusobanurwa nimpinduka zubushyuhe bwo hejuru, bugena ubunini bwingaruka za Rayleigh, guhitanwa, hamwe no guhuza EWNS. Nyamara, nkuko byasobanuwe na Nielsen et al.22, hejuru yubutaka bigabanya guhumeka byongera ingufu zubutaka / uburemere bwigitonyanga cyamazi. Iyi nyigisho yemejwe mubigeragezo kuri microdroplets22 na EWNS mubitabo byabanje8. Gutakaza amasaha y'ikirenga birashobora no kugira ingaruka ku bunini no kugira uruhare mu kugabana ingano yagaragaye.
Byongeye kandi, amafaranga kuri buri miterere agera kuri 22-44 e-, bitewe nuburyo ibintu bimeze, bikaba hejuru cyane ugereranije na EWNS shingiro, ifite impuzandengo ya electroni 10 ± 2 kuri buri miterere. Ariko, twakagombye kumenya ko iyi ari impuzandengo yishyurwa rya EWNS. Seto n'abandi. Byerekanwe ko amafaranga atishyurwa kandi akurikiza igabanywa-risanzwe21. Ugereranije nakazi kacu kashize, gukuba inshuro ebyiri hejuru yubuso bikubye kabiri uburyo bwo kohereza muri sisitemu ya EPES kugeza hafi 100% 11.