AU2013202332B2 - Anti-bacterial applications of poly-N-acetylglucosamine nanofibers - Google Patents

Abstract

Described herein are compositions comprising shortened fibers of poly-N-acetylglucosamine and/or a derivative thereof ("sNAG nanofibers") and anti-bacterial applications of such compositions. The sNAG nanofibers may be formulated into compositions for the prevention and/or treatment of bacterial infections and diseases associated with such infections. Regimens employing such compositions are also described. NAG sNAG ss 05 1 3 0.5 1 (br) shAk p-Akt Etsi Control $26 C. pGicNAc (sNAG, Taliderm) (Fibronectin R) Ets1 IL-1, VEGF, Defensin~s Other Target Genes Fig. I

Description

ANTI -BACTERIAL APPLICATIONS OF POLY-N-ACETYLGLU COSAMINE NANOFIBERS [0001] The entire disclosure in the complete specification of our Australian Patent Application No. 20 11239466 is by this cross-reference incorporated into the present specification. 1. FIELD [0002] Described herein are compositions comprising shortened fibers of poly-N acetylglucosamine and/or a derivative thereof ("sNAG nanofibers") and anti-bacterial applications of such compositions. The sNAG nanofibers may be formulated into compositions for the prevention and/or treatment of bacterial infections and diseases associated with such infections. Regimens employing such compositions are also described. 2. BACKGROUND [0003] Currently, antibiotics are a standard therapy for bacterial infections. However, some individuals have an allergic reaction to certain antibiotics, others suffer from side effects associated with antibiotics, and the continued use of antibiotics often leads to a reduction in their efficacy. In addition, antibiotic therapy often leads to the emergence of antibiotic--resistant strains of bacteria. Accordingly, there is a continuing need for new anti-bacterial agents that are effective in fighting infection without generating resistance or reducing the efficacy overtime. There is a need for non-antibiotic anti-bacterial agents that can be used in clinical settings, e.g., in the treatment of infectious diseases of the skin, digestive and respiratory tract, and in wound treatment. [0004] Wound infection is one type of bacterial infection. Wound infection is a major complication, especially in patients with chronic disease such as diabetes or during imrmunosuppression. Such patients have disruptions in appropriate inflammatory responses, including the migration and recruitment of neutrophils and macrophages, which predisposes them to increased infection (Singer, A.J. and R.A. Clark, 1999, N Engi J Med 341(10): 738-46). In addition, bacterial infection can lead to impairment of wound healing and sepsis. Given the ineffectiveness of nany current antibiotic treatments and the increased prevalence of antibiotic resistant bacteria such as MRSA (Methycillin-resistant S. aureus), new clinical treatments are in high demand. 1 Si N1. RY 10005 In one aspect descibed WKe 1l: are methods for tzeatmg andior pro elmitg a hacieril mnbetioni) and/i or ds.aes associated ith or caused by a bacteiM mfe ton a subject. 006) b) certain eni dimntic&s deteribed herellt are oethodc for u\eMting a baterial infecuon mn a Subjet cOmprising topicdily dmniriga compii~55OnI~0 comprong NA\ T tnolibers to a. 5ubC etIn somne embodieus the iIblec 1 d2 ifagoXsed with the baend! infection or dQLK5I one or mlr t TlffOms of the bacterial 1ouct10 '1 10e htds of diagznosio h'bactenanfctronand jSypost 0"actenial WtAo are those known ml the art or deen <dt''h " ihebeIteral ied en mat be a ski ofeetion, a gasoinesoal infecton, a. res:piratoiy miecuon, a ununary tract oftection, a reproduct ie tract mntection, or infection 0f ant ote Or orsnSor tuscu hi the bou of the *jeel as do'debd hare r oe embod on lnt, the infer~in is a nosocomial iceirn, an MRSA ofteenioit a Psendsoen infetonioraC( dicmid nf ectaio 100071 in ertian emb adentsdsrertnT hr ine ml forreatng and or presatog a dis.dse asOventCd n illi a kictenal mfeton r1 d dialt it'adid t a sub IjCt't COmnprliin opiedlky adimrnlirtring a conQAsitiO I & cmii\ing N \C PiOtibers No Qh in. siih js t h i 0n1 Nuch embodimiem, the method I s N eaamg and or proer nn a diNease aw mecated vu itl a harienal infetion i dnotor 'mtbidiknt the method in\ OlveN treatine Jnd or pr ennge a discd se asocia UIe w it 4 aWt'ril imbadance, tor example, an ambalani e in haemuri' ri ebrta as deseiled rierctinhscra 0(10 Cnbodiment, rhe: metheds mnWo e treat ng in Qxlstal uz aeind tntectio0 n Iom \of theUw emitodinist 4111ubject t bercTated iS digpfOMd wnhl " disedise aisioctated with a buetul ofitctuon tor displays oat or tnore sy mptorns of such di ease, In other embodimem\ the subicen to be: nreatedl is diatgnosd with c ivswaso scinated w ith a bacterial imbalance or displays ine or more symptoms of such iealance I he disae may' be a ski disease s gvtrointestin'd disease, ad res-piraturv dikeae, a urmarn tract disease, a. reproductive tract. bsease, or disease of any other Or gan or tisse in thie boe of the subiect as described herein. I1 -ome Ceu0dimenta the dhN~dv i'n J sin, disease tsr a gatrnlteatiadl d1beC .se h0 oC ormtodimemo, the disease 1s as eiated v. ih a i0% oslmoraa intecilen. an 1I($A iniebo01 a 1SV Od 1 d' .54(Oflk 4C Ihtetii 01' itn L. 6,lkW ilieu'd id ni ita W008 in. somn embodimenits deribed: thereirare methods eo r pre-etng a bacterial inttioun and tor diease assne1ated u ah a haer ial etion compring topically adimmstermg a c1omposium comnprisg \AG nanoib ers to a ueht in some emboiments a comlpoSnin cornprisig sNAG nanohers is adminsrered ta su b jee at high rish ofa bacterial mnjection to prevent a disease asocuated with a haeerial miecuon. in specific mbodimems a nO~flOStifln conltprnu' A sNAG na1tIlers is administered to a vibict wilta wotud or a Micbhet who has undergone a surgery in one embodiment, the composition i animstered to AM iinmfunionflproumed subject bn some enthod.ments. a cornpositilon conmpl nn sN Ac nano1ibers is administered to a nwrind xhere the sound is at hi risk of bate r ia mfection. In is an opnc wound, the open woud may be a nmhot wound ritincture wo0i14t a 1acemtioto wount, ant al'aaion, a ct, a pentrtilon wound, a Sorgircal ouno., or n\v other w ound In certain enmbodiments t vw ou may be a puncture wound, tor example. a puncture wound that is caused by a hemodiai.sv p ocedule or a catheterization procedure In suchl em.tmnf h rtjet to be treated may lhave neeU diagnosed with a heP10dodiaks related or cnrheteri7atio-relaRted onetion. in One ambodniilt, he bacterial ifection and/or the disease asocated with a baeil infecton to be reented by a sNAG COnpotiiton is not >n a wound ( . an open wound), or is not ascuated with a w found hn one such enmbodient, the bacterial fiction and/or the disease ssociated with a bacteria infection is not at the site of a Wound (e.g.. nti at th sie of an open wound O~O9 1 in omeeboiens dsri horcin are methods ftn tient iabceilvifce ouud in a imdao n u iirising G nianoibers to the wound site in a sub ect In some embodiments te sub ect to be treated i dhurnosed with a bacteria! intect!on ot di spl'ayS one or moere svy mptnti ja h bacterial illfrtion, in certain cmbodimems, the wnund is an open wound I he open wound ma be a gunshot wound, puncture wound, a laceration wound, an abrasion, a cut, a perunration wonnd, a surglal w ound, or an n ottwr wu nd o in certain. cmndments the wound nfa) be a puncte woGund, for exanple, a puncture wouund that is caused by a hemodidlysts procedure or a eatiheterixation proer.' In. sudh esllbod~llents, the sudiet to be treated may hav e been diagnosed with a bemodialy sireided or catheter1aow related netion |00101 Bacterial nr Rions to be treated or prevemed using the methods described heren include infections with bacteriu of One or Nore of the loi aenuses Bordella &i'0u, .xth'e/Ia. (am;:/barr. (t e tamn da Gmandi C/taw AL 10. toJam.n Enwencthas.t L clh lOelb/,iaT~i'an hteild. Th micpit'a thib etW, Lrai ne/hr Luf.e'r spi'a 31 Shigela *Staphyococcus trptocuccus T rvepnan, TWVi, and Yersinia, in some embodiets, a NAC COmfpositi~ o may be ued to treat and/or prevent a disease Uaociated with an infection by bactena fion one or more of tne listed penuses of bacteria, or one or ore aymptoms thereof. 100 B r in iAos to be td o prevented 'ing he method& described herein also iue ections w e Wc of o oothefospeies: wt dinc 9$ 1ke4'eUpOC n s s una ODI ,Bae//a cn/.Tecc/aen T lWars COI Mfslub' A; nULWAA M 0uaW Wi n ba h"'. tub die Mt'ias p nCseon L?;'ue ~ ~ ~ ~ ~ ~ b bu/ia )NO>M 0.4 el5 ph' -f V 'fe k;ti dimri otU )I ? iSUMJWne a/ N~OWa&orino or, Ooc qMquU PS'm 4vhr/e tcdoUSicas(%lC cftDS foolad an co srm rCn vants. NA c/o pason a o m uOed Tor ad p t d> >~~f >lfhfl i N? \$n10D''/U Mod=s'i~tt a hg/a5PC.V4AVQQkU ue a dease asso edwth anection b nacteria rom one mo othel stespece t methods desrnbed heremn. tn MRSA ofW. tion, a Pw tet nfbcio or o ./ C/ea uieetot, ln some 'emhodimer1ts a si\tk euniposimin roar ie used tO treal Ut~d Q1 Prevent a discea asociated \MRSA infection. afhearanona inf'ection, or a ( d/ifcve infection, or one orn mra Symputais thereof Syuptom thereof W13 In certain emod'intos the bactehai infection to e treated or presented using the mctiods dc wribed herein caused by baceoa that are known to one ot ordmary ski m the ar to be resistant to a tan dard anti~aetenal therams for e:<amle resstam ton or mi aibioncs. ir one emabodiment, the batenal ieetion to be tre d or prevented uing the methods dscNribed hrein is MRSA. e a noc maMRSA hn some embodiments a sA composition may be used to treat and/or prevent a disease associated with an infection by bacteria resistant to one or more antibiotics. In one embodiment, a sNAG composition may be used to treat and/or prevent a disease associated with MRSA, e.g., associated with a nosocomial MRSA. [0013a] In other aspects, the present invention relates to the use of a composition comprising sNAG nanofibers in the manufacture of a medicament for treating a bacterial infection, treating a disease associated with bacterial infection or bacterial imbalance, inhibiting the development or onset of a disease associated with a bacterial infection, inhibiting the recurrence of a disease associated with a bacterial infection, or treating a bacterially infected wound in a subject in need thereof, preferably in accordance with the methods and procedures described above. [0014] The subject to be treated using the methods described herein may be a mammal, preferably a human. The subject can also be a livestock animal (e.g., a chicken, a cow, a pig, a goat) or a pet (e.g., a dog or a cat), or any other animal. [0015] The sNAG nanofibers contemplated in the methods described herein may be of varying lengths, widths and molecular weights as described in Section 5.1, infra. In particular embodiments (i) the sNAG nanofibers have the average length of less than about 15 pm, or (ii) the length of more than 50% of the sNAG nanofibers is no greater than about 15 pm. In certain embodiments, the majority (and in certain embodiments, at least or more than 60%, 70%, 80%, 90%, 95% or 99%) of the sNAG nanofibers, or 100% of the sNAG nanofibers, are between about 1 to 15 pm in length. In some embodiments, the majority (and in certain embodiments, at least or more than 60%, 70%, 80%, 90%, 95% or 99%) of the sNAG nanofibers, or 100% of the sNAG nanofibers, are between about 2 to 10 pm, or 4 to 7 pm in length. The sNAG nanofibers of the described length can be obtained, for example, as described below in Section 5.2, infra. [0016] In certain embodiments, the sNAG nanofibers were produced by irradiation, e.g., gamma irradiation, of poly-N-acetylglucosamine or a derivative thereof. In some embodiments, the sNAG nanofibers are produced by irradiation of the poly--1--4-N acetylglucosamine in the form of dried fibers (e.g., at 500-2,000 kgy), or irradiation of the poly--1--4-N-acetylglucosamine in the form of wet fibers (e.g., at 100-500 kgy). [0017] In certain embodiments, the sNAG nanofibers are derived from microalgae. In another embodiment, the sNAG nanofibers are not derived from crustaceans. In yet another embodiment, the sNAG nanofibers may be derived from microalgae, crustaceans (e.g., shrimp), fungus or any other source. 5 (followed by 5A) [0018] In one embodiment, the sNAG nanofibers comprise N-acetylglucosamine monosaccharides and/or glucosamine monosaccharides, wherein more than 60%, 70%, 80%, 90%, 95%, or 99% of the monosaccharides of the sNAG nanofibers are N acetylglucosamine monosachharides. In another embodiment, the sNAG nanofibers comprise N-acetylglucosamine monosaccharides and/or glucosamine monosaccharides, wherein more than 70% of the monosaccharides of the sNAG nanofibers are N acetylglucosamine monosachharides. 5A (followed by 6) 100191 in enain enbodiments the 3\k\i nanovhrsised. in dhe methods desribed herei do not have an effect on bacterial growth or surmval of Stphoecu auroi baCterial cultures M n'v or subtni ally have no etfeet on bacterial grow th or s urwi of'Stph rf u<t au a bacteral cultures n vlrot in some emdod.ments, the W\A% nanoiers reduce bacterial growth or surx is a] of bacterial cultures <n trr by eAthan i lig, 07'5 lo - lg 3 o 2lg 0. og, We w~x he'n Sahwdoc -u ace habeLtnal culftres arc treated/ineualted with th mN'% nanot1 er W 1 rom The tests fbr the etThct ofi\A NV rt 01 ) baceri ait\nt'h or \U'\ il Jo and the eval ahion of the test resuhs are descrbe, Or example. in Section 5 1 &unpse 2 te Netton 6 "2 ' ) and f linre 1 .12 k ire [900 In certain embodimems the s\AG nanonbers used in the methods described nere ,are non*react\e in a bideom lbit\ test or tests For e\ample, the tNA h nanotibers used in the methods d nbed heiem may b non-reactive whea tested im an oltom test, ar itramuseutiar implantationt test an mutraceutaneous test, or a. systernie test. in some embodiments the compositons described hereim are notneacti vo when tested in an e I tiam test, an ntramuscular plantation test, an intmeutaneoua test or a systemic test. in other embodiment the sNAG natiers wsd in the luethods described heren have Grade 0 or Grade when tested in an Chin test, an imramuscular implantation test, an otraentaneons test, or a systerme test hi yet another embodiment the s AG nanofibers used in te methodS descnbed hrein are at most dldly ractime whBen xted i an ehbtkon tSt at) imrauneui itmplantauon test. an iraculaneous teo or a systeuc test. i one embodmnL the ' A nanofibers or eemipostin comipr1m a such nohibers ar norneacrtio e a dtterinsed by an mitnmseular impiantalon test in rKtam embodiments the compositons described herein do not cause an. allergeu i-wtartion or in irritatiers e g., at the site of: application. in other emboiment the compositions desctbe d neremn cause at mositammld aler gemic action or atumdd 1rrrtauon , e g at the sit1 o aptcaurmlt 0021 -rhe contemplated modes of admistrAi ont of the cornposlions described herein are topiCA eg opcal on the skin; topicalat the site of a wound. a surgery, a bacteria infection. or a symptom o an inteion (eog, a velingh and 1opkai to a body urfa . ch as the mucous membranes (e g ,aira anus throat, eyes car, or the surfeC of other tisues. In certain embodiment the sl\AG nanofibers or cotnpositiots eomiprising such nanotibgrs are formulated as a. dressin . a bandae a mat. a spray, a hq ni, a suspension, a membrane. a 6 ptn der, an Oimmenfcft, a crean, ai pa te, a suipposuory, Or a pl in SOmei embeunents. mesACG nanonoers or compositions cornprin ig such nanotibers are formulatec as a cream, a gel an oint ment, a membrane, a. powder, a spray, or a sappos1tory (1221 in another aspect, descnbed hrein are compositios br uei the methods desenhed herein inapei etlc embodiment. the composins ComprA SNAG nanoibers In Cemin embodiments the compositions described herein comprise sA\G manofiers and one or more additional active inirndentsusefu m preventing and/or reatine a bacterial infection, a disease ascuiaed wth a buewrial ineton or a symptom tereof in some embodhments the aditonal active ing'redi ln is an antiharterial ageL. Suc~h ecdditional alti~bacleria agent mayv be an antibiote In anotiter embodimeent, such addi tional anti-butterial uge'nt is zYine, I vet another embodiment t compositions described herein do not comprise an antibiotic In vet other embodimem the expositions described herein do not comprised ay addithin "nh-acterial agent In one embodiment, the copositions described herein comprise the A AG ntibers as the only atv ngredient and d e not comprise any additional active agredient: 100231 in specific embodiments, %omposion comprises the sAG nanoibers and an ,mnit1ib.i e, xamp nioi that can b tased i the composinoVa of the imvemtuo inc bide maicrmhies (c e rvythromnyein, aaithromycin), amninogly'eosides (ett., aimkein. gentamicin, n~Ieom CL, stre ptomyin), cephalosporins (exg. cchdroin, cefactor, cetarn e, eefeimct fluooquioloes tstu lwotloxaelt, lVOftainC~f),fO peicitins (< .. penicdlih, ampielI in, amoxicilltin). tetracyclines ( e, tetracycline. dosyceelna i andor carbapenems (e., mneroupenem., iuipeneni The sNAG nanoI~hors and dgnnts dscbed herein max be nsed i such compoitions in some embodiments a composition comprises the sNAG Panofibers and vmi agemt effective to treat or prevent or emmroivl used to treat or prevent auS ttui m sectionn, MR SA iect ion, a PwndAanona tmicetion, or a (. dilwwue inicction (cC.s an ant ibzotiez effctise against or cmmonly used against such infedtionsf (24) in. other embodinents the eormpouit'ou described het em are' adrmm~ee idn conincidon wi(th one or more additional anti-bactersal age t s or any other 6i able thn's'p) . n Some emnbodimients, the additonJ.l anti-buaterial agnt or therapv i-s 'n anniott' ('e a a anardard antibiotic therapy for the bacterial infetion or a di se'we assc ated. M ti a tcterial infection to be trea tod, as IknownU in the art or dcntCibed herem)t in Ati y ethoduimtims, the add ition ant>~ bacterial agemi is an sent effeettre to treat. or prev ent or om nyusdto treat or prev ent an li loomot ue cure i otection, \MR Sv\ irdectimw a PwHencn infetom. or a C daidc itfectione.e.g . an anibiotie efteethv aaainst or commondv used iatmt rich turn'citor In some emhodmintts the additional iherapy isadmimistered bef ore, sirmuhaneoust uwith or after admaimstrut om of a s\AG nanonber cormpostion. in yet another embodiment, the cornposatoms deseohecd herein are not administered mn conymuetamn uith any other therapy ca.not adinistered in conmonerioni in an anti oti 33 Term inaI*foC OQ2Sj As used herein, the terms tNalAG nanoiher." 'tA, Taiidernr or ''a vulyned (fornnerdy known as "Tal idern$"s are used ierchaungeably o refer to shortened ibersa 'poly44 aectv tgiucosam.ne and nor derivati yes thereof 00261 As used heretn, the term ",ibot" means a ranige arun a grven udue whereina the han-tint? value is the same or iknatmal the sam-e ie cw fifhn 104 S, i or INo asthe expressly recited vidue. In one embodiment, "about" mean w; thbin 10% of a ven vp ue or r ange. ln another emnbodtrn nt, th ter. ni "about" mesms o'thin 5% of a given uduie or range. in another emboiment the tern.mbout" rmean w itni 01 o ot a giv aiue' or range, |00271 As used herein, hierm itise, "diarderr or "condition" are usMed initerchangecably to refer to a" medical condion mn a sudjeet. In a species emb~owlimem,. the disease is th padholoiui ate associated n ith or caused by a bacenial infection. [0028 As used herein, the P1rm "bacterial m fction~ means the munvision by rumiipl ication anidior presence of bacteria m a cell or a subject. 0Q29 gAsed herelin th uei ma refers to Iog 0030 As used herenthe tcerm 'therapie" and khap can refer to anyl prtt)cIt4 method(s) composinons fotrmuhdions, ad/or agents thatcarne used inth preenton nd/or treatittatof a hacitrd' enon or a symnpem or conditionasoeiated therewith. incertain cimbodiments, he totnrmteany' refers to a sr\Gw namotieu{ al or a. pharnnaceutcal cooit iton corimpns ing a a' AG nantotiler(<. hi other tembOOnen~itS, the tenni "therapy" reters to ai therapy other than a s\NAG nanofibert s) or a phatntneutiaia eompostion comprising a s\G nanoiberb) In speitic embiodlmntsitt >' additional therapy"'and '"additolid I tierapies' refer to a therapy other than as > AG nanofiberN or a pharmnaeeutical enmposioon comipr ismy a s\NAG nanotiben s) ha a specific embodurment Phe therpy m incues use of'a sNMG nnoiberNs or pharniaetibeal eornponlion Cltnnti ntg a aN \U i iioutertsj as an adliuat...euf or 3K xarmpe, uing a sNG nAnti ber o odoiion in Crin-i nith adig therapy ih a n antibitic, andor Oher therapies usefl in treatmentander prevenim bacterialinfection or a symptoml or conditions dsociated therew ith 10031 As used herein. the ter n "ef'etive amount" in the context fa dmnistenmg a KNA another composiion to a subject refrs to the amount of a sNAG idnotIber tht realts in a neneetdat ortherdautclc [itn 1i \.cciflne eltbilblwand "effecvle amount of a sNAG nanoiber refers to an amount which is su'fiTint to achieve at least one two, tree, Oar or more of the toI010 mg ef ects: Qi the dwen0ce of a bdral in fecttionL Q) ho eralof Ofone or Tuore s)'m1to01s Jussecated therewith, (iii) the rdi'.ltan of timu reoutred to clear a bacteril infection i) the reductin or amel oration of the seVra *4 d bctenal infection andfer one or more symptoms associated therewith; v) the reduction in the duration of'a bacteria section and/or one or more srmptomis aswciated therenwithi the pre~ eniou. o dela of the gener action of a resistant strain or strains of bacteria or reduction of a i nib of' resistant strains o1' baera generated, i\ IT prevenin in the recurnence ora acteri iet aAn and/or one or mote symptoms aS 1ated there th ( \i) the reduction or chinination in the bacterial cell poplation Ox) the reduction in the \everiy and/or duration of a condton caused or dasoetad uith a bacterital Wnterioil U') the trduethon in hospitahzabin of a ubjet (ai) the reduton in hospidaizatn length; hxii) the iners n ite survive al of a subject; (xiii) the enhancement or onpwosement of the therareute efetanother therapy: 1xiv) a reduon m mortality; (o the redAcuon or CHmination in the spread of the bacteria from one subect to another subjet, or One organ or usnu t:aother orman or tinse; (nvi) the preventon of an increase in the number of bacteria, (xvi1 the prevention ol the development or onset of a hacternal infection or one or More Nymptorn associated ther'wth \i) reduction in the nmber of sy)ofms dasoerated CAith a bacteriJal infecton. (\'1%)e e notion0 in the duratina and/or seerity of a condition cused by or associated with a hhterial fetiKon, (mx) the MNhih=uor r redetron m pinduction of a seial toxin o tons awecated w hi a acterial infection; (\\i the stabilization or reducton of'mnfammation associated wth a bata int on 5 m (u>m th.e induction of the expression ot'one or more detznsin proteins and or deferisin hkc potelnc niN i the induction of the expresson of one or more anl1-ke reejltore ( wix the itdu"cton t' the expreston of oneor more proteas that are beneflcid fir clearane. or reduction in a bacterial infection or one or more symptoms associated thereuwith, (xuri) the reduction ma 9 orTgan falUt acted with a haeedi in X.in Oa disease dwsocited tlerewith; 10b \ ) the prevention of the onset. de\elopmem or recurrence of a condition caused hk or asoctated with a cind in& etion. and or ixxviil) improvement in qualty ot le a tsessed by meohofis Wed known t a Questionnar In specific en dhcine dll an ceA L live amount" of a N AC nanofiber refMrs to an amount o05 a sA nanofiber compos1o specIt r e , m 100321 As used herei the term 'iderdv humanrefers to a humn 65 years or older. 10331 As usaed herein, the term uman AdM oeltrs to amatha i years ole 1034 As nse hre t term uan child ers to aurn that year o t old, [00351 As used herent termdhmn. infant" refers t a newborn to yeol ear 0036 1 As used herein, the term "premature hutn infant" rfexs to a newborn to I year old year human who w as bor of les than 37 weeks :estational e (e., hfhi 37 w weeks, 36 weeks 35 weeks 34 weks 33 w eeks. 2 weeks. 31 weeks. 30 wes s weeks weeks or en than neeks of pIrgnanc 4. 10(371 As used heren, the term. humann toddler" refers to a human th is 1 year to 3 years old. 00 As used herein he term taorit refe t greatean 5( rnleding g 503 j&039 ~ ~ 7 Asusd erin the er 'au bjeet)'. ad ptearusd ivntrchamtajl t o r wbrt an animal (eal. cw, bon i Mttnk phif.w ch n Ar ""y quail Cal, dog. I"ot vhitY "nine pi, efe. in a spec.fe mo1dment hitbj i N n nad such as a non-primae or a prmat dCiJ. , a a.In styee le tmlv'ud ienb ille slbject is a huinan. See Section Ci Can D~4 r moftett information concerong patients created in accordamee nwith the methods provided herei 10040 As used herem, the term '1w eAppeion in the contest si espieKiin of, acne (eo. based on the level of protem or reide produced by the gene re"'rs to An xr Ou tha is less than the normalia" expression of the ee hn a specific embodhnem, '1ow t~nisitonA trors to cresson of a ane that is less trn M% ic.s than 50% le than 75 less than 70%, esn tha 65%. ls than 60% less than 55% leS than 50%, lss than 45% less than 40% leW than 35, less than 311. less than .5%. o les than 220 of the "normal expression of the I0 geniseh U0 mininriuecit fie-mhod nient."low exosatn refrs t. espresio of a ee that is sut2 O~d 5 bout f5 d about l foid about d aom 44d about 3-ddabout 2 fdid. or hot 15 fold les han the "nomal" exprssion t the gne 4 BRIEF I)ESCR WTION OF FIGURES 100411 figure 1 Nanofiber stimulate At 1 acti\aonL an upatream -guItor of Eta Wernern bit anals of phospho-Ak!t re pons o NMI ,md AG shnnuiation of sr samed FC, (1B) RT-PCR analysis of K infected enher wih scrambled control ("8CR')or shRNA lentrtiruses and asneed for expression of a I arid S2n as a loading control (0 Schematic of a sina transduction pathway transducinas al from sAG ruaotibers to At 1, Et ariand De lmsiw [00421 Figur 2 D yed woud healing in Akti null animasis pattially rescued by denrn treatmen ( Representate-tnages of ded WIT' and AK.T dnulH nice wih and wihou teatment of Taiderm (H) H&E staring of representative mouse skin seetiomsftom day 3 wounds f0043 Figure 3 sNAG nunofihers stimulate egvtYnae and defnsin depression in pnan endothehal eels (A) lmmumhistoehemsc y of - C treated with or Aithout sNAG using an atiriuhdy dn'ee(ed against auneiern (\ I IA \how ingt nacr tr ent of 1h insults in Th secreion of sdefensns icrum strtd treat-d n h N pg mi or i10 pg -mi -AG) )00441 Figure 4. sNAG vanons ibErnulate dee'nis u p r.iary endmthial cedN> ina n At1 dependent manner (,\l und (13 Quantin l R -P ain'lses o 'cuma sta er E t (s') create nith or w uhout SN\G ' a' n Vh or ws udhoot PD059 I MAPIS whibitor, -PD'. Woarannin ( P13K inhibitor, -w t'n') or iited Awih a sci e outrl ( CR''. cv ,Ai ('AKTI> shRN\ lenti\iruses -id as1esed lor epresion orthe genes vindicated 100451 Figure 5. sNAG nanonlxiresmnuke | defensir 1 api ewon m rnus i0erano0y1es (A) inmunolluoreseev italingS IWA defAnsin 3 (\-R 1 s briuht stailninl in the upper ahot hand panel; se e. g, thick Shite arrows) and involucrmi an nboies of pwdfh embedded mouse -as w found se-cton- rom \ and it] null ammua on D ,ay (B) Quamti fira son of |ldetensin A irnunofluorescent staying usig NH il mage) software (\-XwI adenu, Akt , Akt i null) (C) immunofluoreent s'tamm of W and Ati null treated and untreated keratinoetes with VDefenoin 3 (visble as bright signing; se, eg., thick white arrow) and TOPR-3 (nuclei staining: see, r thi wlme arrow s Notice the imcrease in f Defensio 3 stainign in WT and Al I Faliderm treated w pounds 1(1(4) Figure 6 .Akt). dependent transcripnon fhctor binding se Schemati of Akt. depen dent transcenption factor bdgst sing Cienomtatix sotuare, 500 np upstream of the transcription start site was analyzed ft onserved sites on the mRNA of DFF, , and 5 (FTS black ox ats K] l1Dastured orvals; CR1 B n it' ovals, Nh K Bcheckered ovals), 100471 Figure 7. sAG treatment reut\ in expreson and secretion of defensin /n ra. iA) RT PTR anayss of senm starved SS prnmary endotbebal cells treated oth :A \G (50pgml fo the tnesxidicaeuwd n asese fur expresson of Vdefensio 3 and n~def~osm 1. (B) mnmunofluorescent lhnent of endothe w els either serum starved (untreated) or treated with sNAG nanofibers (10grd for sr Antibodies are directed against a-defensin 5 (MIT, upper lef t hand panel), Cde un i eis Red, upper arih hand panel). udei are staged with TOPR OK (Blue, lower lett hind panel), L ower right hand pamel repesnt ine r eray (C) immkohtoreseent labe ing of inoeges (f kat that are eiher enos at arv ed (untreated) or treated ub tSNA\ nAnofher\i 19 m for - hours i Antibodies are directed against o-def.nsin 5 (FITc upper lei hand pandh, p-deensin 3 (Tomas Red, upper right hand panel Nuclei are stained w ith TOPROK) ( Blue, iower left band pant.) |00481 Figure 8, sNAG induced defensin epreu4nn i dpendem on Aktl. (A) Quantitatie RTKPCR anal yse us n primers directd agnmt ode in m irno total RNA isolated from serum starved endothchal cells tiad o h 01 without IAu tor 3 hours w ith or without pretreatment with PDPO%t (I"PDNS0pM),aw aornnin ("\\TW''( I0nm4). Quantitaion is relave to te S protein subunit. (L) QuaWnutaun of pdefensm 3 expresso from total RA isolated from vernm starvd endothelialteils ireaed with or without A fobr 3 hours with r without PDS)059 (50pfou wortmannin (10%rn) and show n as relative to ~ U (t W western Blot aalyss of phoopho- MA in serum starved endothba cells (35) stimuated inth s\A or the times indiated. inme udica.t where lanes hav been mov ed (D Quantitatv e RI0-PC R analyses of serunuvstarwdv endothela ces mtoected w itha: scrambled control (SOR ) or t i shRN lent iAss treated wyith or w ihout sNG and assessed for a-defn&sin 4 expression Quaniation is shown relam to S6. (1) Quantitation of 3defensi 3 epresson rom total R\A isolated front serum starved endothelial ecls infected with a sermbed corol (SR) or \kti shRNA\ lenti viruses treated uith or without sNAG. Quanttuin is show n 12 relative to S2n All experiences wre done n at least r pheae and repeated at east three independent tines and p "aus are shown, 100491 Figure 9. VAG imued def'nsm expression rie requires Akt A ) Pa ratin embedded xcctons of cutaneous wounds harv tested on day 3 post w ounding Wrm both Wi 3f and Akt1 mie. Wounds were either entreated or treated with sNA membrane lmmunotluorcscence was perform d usug anibodieb dreted against Ketnn 3 (green, vsibie a srghtadining nt the upper right hand pi, see, a. wIte t IN arrows), Inloluero n i Redt and Topro (Blue, uclei staring, see, e y wht tKn arrows> (R) Paraffin embedded SCenen from \\ trevated with sAG harvested on day 3. hounofhuoreseence was performed using antibodies directed :amt T defensin 3 (gren, NObL w\ iniht stamin: seec, g., thick wIe arrows), involuerin (Red) and Topro (duc, nuclei smaing se, c g thn wh0It arelw Thns lower nagoticaion. 10 P meluded to her luse ae eh vpidrnna laye eirs defesn 3. Scale barb 50 gm Wt Quantitation of a idenein i expression rom paraffin emiwdded seens w as performed usmg M IIl imanel software. x nperiments were repeated thr e mdciadent times and p values are shown 100V01 Figure 14 sNAG treatment increases wound esosure in wild tyrpermic. stanm g of wound tsue sections dernd from 05B16 wild type animals zither untreated or treated i n \NAG membran, The day pos-wound is indicated to the left of each imc The sobd hack ne follows the keratinocte cli layer ndicato v O indicate th" margin of the w ound bed. f0051 Fgure 1 sNAG treatment reduces material ietion m an kti dependent mannerc (,\ Isuc gram staining of reus infected wWounds fronm \I' minK \\ micen ete wounded oJAW a 4 mm iopsy punch media w ondmg niee w ere inoculated wyiini a 10 9 e fu m i rainu tea posmincon, ice in the treated group were heated x aX Ialider Skin samples w re taken 5 days pos-trcatmenr And ctioned t0r analysis. "imue gram staining was perfoneol Dai k pur)in staining indicates gram-posiN e bacteria and neuronhds that he egulfed bacteria, Secions under 20s amd 40 magnication are shown ( ) Tssue gram Sawing or paraffin embedded 5. 6re incrted wounds from \'T aid Akt l null mice (ni: Inftd wounds were eiber untreated or treated with s\Ai membrane and w ound beds w ere harvested on day 3 and day 5 or analysis Part purple staining indicates the presence of gram postave actera in the wound bed Black arrow indicate examples ot gram positive staining 13 Note the accuml nation of pwsmx e. stamii: m manreated WT that is tackimg in \\T anmias treated wit nNAM Scale hars pm, (0( TO derned fom day i posaounding wre quantitated trom atn infected nouad1 ng bot tred and untrated WT in so):d At1 mice (tvt:3)\Wild type mice that nware N\ \V s heated show a significant (pc p(1i decrease mo baeria load in the wound beds as compared to Aki nud l annals All experiment were repeated ihree dependent tmes and the p adues are shown. (DI CPU ouantuatd fom ofeted w ds at day 3 post wonding in a simita fashion described in (C NA\Q treatment of infected wounds shtow a signeant decrease mi C tof both WT and A. nl anmmats on day 3 but the WI anmals shon' an approumae 10 fod difference compare d to a o iffidJ'erence m6 Aid I animals. (F) Quantitation of (' 1 s in 5 rw'euesc'ukur"s that wer et'ithei unteated or trae w ith various amounts of aN\AG nanonibers. Each evpci mnt w as performed three independent nes and p \values are shown. (I d tine gran stan ni of N r&'tnfev ed wounds har\vestd on day 3 post wound from WI' T n 31 that wecre treated wth or without & defensin 3 pepide 1O M Nte the dee in cm nam potoletammi m ofeted wonds that rtrAa wt nh defensin 3 peptide t0) QuamitaGon of CT trom S. aaewns infected W' mice (n ) un 'eted svith or withot 'tdet'eno xi 3 eptide lnfected woumd, that were treated with pertude 4heW a irnficant decrease p <05) in CI1 Scale bars - 10pm Bach experiment was performed three idependem tmes and p \""U4 dre showN 100521 Figure 12, Rapid mdians ion ''I nreasrin by A treatment ofS aun. infected wounds. (A) PaRaffia e cddcd tissue sctiio mm aures infected w ounds harvested on day 3, ere subvected to mmanoiuoreacence usms annbodjes directed against p dfosin 3 (green, a"iibl as bright stain in the upper rigt hand o nmel a'd in the lower pane in the middle' seIe n- duck nhite aoi a L yotrin (md to riairk th. tttnocyte layer, and opr (bh, uclei staining; see, e Og tin w lute anrout roi boh sN \t 3 created WY (n:::) and untreated \VT mice in-3 t Non speeifc sming of keran is indicated b5 the no primary control w hich wa ustamned w ith secondary amtibody only Scaie nur 50pm (3 Quanttation of V defensin 3 expresson from paratun enmedded seettons usb ng N It ImageI somnw are., are iectedl wounds that were treaited with kNtAC 5 how a sigmnIRanm ph rea (ps tO in frdefensin 3 stainig. brciments were repeated three independent times and n x alues a e shon. (063) Figure 3 A ntibodies against pldesin m 3 impedes antibacterial e'eets of j.A treatment (A) Tissue gram staining of par affin embedded S and i feted wounds treated 14 wIth NA Aom W- mice o in ; that were harvested ont Dt , sN\G treated wounds were treated with either f-detbnin 3 armbody or OW ccntrol gOa Qg mibnd MY 1o A \C etesint.d RepresertadWe images Mhow increased aeedl ation avnt positive staining (bnck arrows) in the would beds ot moe created with an antiody dneteId against Wdet'nsi 3 Scale bar - 20pm. (I3B Qntition of CRs rom S. ac' infected WT mice treated either defensmn 3 anibod (n 1 or control UKG antiboY i 3) pIor to sNAG treatment. efensm 3 apphcaton smficatnh increased i(p <05) ( F. 10541 Figurt 14 'N tratnreit ics bactenal infActin by flannw arg sa Mie w re wounded using a 4 imn itopsy pUne , moculated with 5 x 10& I fu m P, "og'ow Onfected wounds were either untreated (n or tted (n with AG membrane (nin) 30 mn posMtectRin, wound beds were harvested on day 3 for analyss cultured for 30 minutes plated, and CF of the untreated and treated nteted w ounds were quantitated sAG trea ted nece show a s'gnifhant Ip ) decrease in bacteria load in the wound beds as womparcd to untreated arnimals 00551 Figure 1 I Ulet of irradition on chemical and physical structure ftl& N e fibers. (A) Correlation baaicn molecular weght of pleNAc and irradiation eve 'Mulation for Jrdiation (P in'raed (OR pectrum ofnon-imiated pGbeNAV slurry (top 1inet p&N Ac surry irradiated at 10 ki (bottom line) and PicNAe slurny radiated at 20(0 ky (middle net (C Scmng electron microscopi (SEA otpilNAc. (D! Scanning electron microscopic (SM) analyses of sNAC [et056i figure 16. p(lNA did not waffc MAoi rate FOr each o time l seiod f i,... at W and 45 h ff dooi for ea ,uof the four bars (on eft (o gh) iw sern tarvation S mi ad pGleNAe NAG) t and 0 p) 100571 Figare ~ (IleNhK protecte human umv hlwe! ndotheial cell (HO) fron cel death induced by s'rurn depriacin Fo each dir pero ~ '24. 48and 72hors), the nt C t f boerum stavaion (S VEG and plN Ac (NAG)at 50 00 a d50 nnaYI [6058 Figra I sAG idued marked irna eae dentigfrac oth five rs fom left to rih s as follows: serum starvation (SS). V iOE. and sNAG at J0 100 aud 200 pugoo 15 f00591 tiur '1. s NAG did not protct IT tfr mu Rcdath indce by ser deprvation. For each tine perod 4, t i and s hOs),he entity for each of the ve bars fom led to rig i o station {SSV fEF aod sNAGat50 00 and200 nm T0060 1h \O nor ha\e diso eroed t hat NAG nanofibers decrease hacterial intention of cutaneous w ountds~ mkected wjth StupheMcol crrW$h imiad P CdQO~m acit-insa, Wuhour heang bound by any Nwcifhe mechailsm of actin data pres-ited ino etio 6 sugvsts that the >mibacterial ffet'e of A G is not due o a direct inerctao of 1AG with the bacteria but K du? to downOstrea) aftect\suCh as for example the repUlation of dbCbnsins by Atl activdato bpecifiUvydata show tha treatment of bacrial c ubares u h SNAG namotihers in 1to does no aIffa baCterial count idicainn that sNAG nanotiner\ do no t directly ihihit bacterial growh. in a snpeelhe eanple described in Secon~ '" and Iustrated in Figure 1 IFNG nan0Ibers do not aVe a direct cettet on g-tot o Swni\ o avwOhYAAvh. Us t'$ J"A The teOst described in Section1 h 3i2 in/m. may be used to test the lack of a direct efieet ofs\NAG nanioheis on bacitial row th or survsa h: this example. i$ tr cutures im utuon St treated with varying concentrations of NAG nanofibers for thre hour, cultur. wer then plated OVeW'nigh at 'mbhat-rialh CAM determined As shown ti Fyrc . 1 no efiet on bacterialgrrn or shveval wIas -1eed. 0461 Th ionventors of the present in venton have found thatt sNAG nanofibers can stimulate expression of defens wich may boost the inate nti-haetri. response, It is widely accepted thatt defensins are important plAyers in innate immnnitv and function i anti bacterial actiites As denontated in the example preened Sections 6. and 6 .... the invCntors of the present intention have found that A nAnofibers can increase the Cxprei n of both e& and -, pe efensins mn endorblieal cetis and 1kvpe defensins in ierat ipoews mi tro and in a wound healing model i r in" vivo, 100621 Further as demonstrated in the 6-emples presented m Secnons 6.1 and 6 ^nfra, but without leiy bound by any special mebarisnofaction, At appears to be tupttant for N AGo-dependent deftnsin expression in \t1M and ino in a wound healing niodeL Consistently sNAG treatment decreased hactrial infectin, of cutaneous wounds inferted with Suphv loccum emu mn wild ty pe control animals but not in similarly treated AkLI. nul 16 [0063] The inventors of this invention have also found that a number of Toll-like receptors can be up-regulated by sNAG treatment of human endothelial cells. Toll-like receptors ("TLRs" or "TLR") are highly conserved receptors that recognize specific molecular patterns of bacterial components leading to activation of innate immunity. Recent work has linked human defensin expression to TLR activation. In particular, stimulation of TLRs can lead to increased defensin synthesis. Thus, without being bound by any mechanism of action, sNAG nanofibers may act as a stimulator of innate immunity and bacterial clearance via the activation of Aktl. [0064] Accordingly, described herein is the use of sNAG nanofibers as a novel method for preventing and/or treating bacterial infections and diseases associated therewith. In certain embodiments, treatment of bacterial infections with sNAG nanofibers decreases the bacterial load in patients. In specific embodiments, the use of sNAG nanofibers enhances wound closure while simultaneously eradicating, decreasing or preventing bacterial infection of the wound. 5.1 sNAG Nanofibers [0065] Described herein are sNAG nanofiber compositions. sNAG nanofibers and methods of producing them are described in US Patent Pub. No. 2009/0117175, the contents of which is incorporated by reference herein in its entirety. In particular that reference discloses sNAG nanofibers the majority of which are less than about 30 and preferably less than 15 microns in length. The sNAG nanofibers described herein comprise fibers of poly-N acetylglucosamine and/or a derivative(s) thereof, the majority of which are less than 30 microns in length as measured by any method known to one skilled in the art, for example, by scanning electron microscopy ("SEM"). More preferably, the sNAG nanofibers have the average length of less than about 15 pm, or the length of more than 50% of the sNAG nanofibers is no greater than about 15 pm. Such sNAG nanofibers may be obtained, for example, as described herein. [0066] In certain embodiments, the majority (and in certain embodiments, at least 60%, 70%, 80%, 90%, 95%, 98%, 99%, 99.5%, 99.8%, 99.9%, or 100%, or between 55% to 65%, 55% to 75%, 65% to 75%, 75% to 85%, 75% to 90%, 80% to 95%, 90% to 95%, or 95% to 99%) of the sNAG nanofibers are less than about 30, 25, 20, 15, 12, 10, 9, 8, 7, 6, 5, 4, or 3 microns in length, and at least 1 micron in length as measured by any method known to one skilled in the art, for example, by SEM. In specific embodiments, the majority (and in certain embodiments, at least 60%, 70%, 80%, 90%, 95%, 98%, 99%, 99.5%, 99.8%, 99.9% 17 (followed by 17A) or 100%, or between 55% to 65%, 55% to 75%, 65% to 75%, 75% to 85%, 75% to 90%, 80% to 95%, 90% to 95%, or 95% to 99%) of the sNAG nanofibers are less than about 15 microns or less than about 12 microns in length, and at least 1 micron in length as measured by any method known to one skilled in the art, for example, by SEM. In specific embodiments, all (100%) of the sNAG nanofibers are less than about 15 microns in length, and at least 1 micron in length as measured by any method known to one skilled in the art, for example, by SEM. In certain embodiments, 17A (followed by 18) the nuomty (an al er'tam embodimets at leat 60% 70%i C.0"%o .% 9{%, *)" 49 A, g ,i" q 90J onr 100A or beaneen :5% to 6 :% to '5" 654 to 75% lo 85% ,5% to 90% 800 to 95 0% to ar 950;: to 99% o' the s0\G naownfers are eial to or Ics thn. I ' , 9, 8 or 7 mcro1ns in len1tH. and ant I nmcon i.n etgh as measred by amy m'diod known to one killed m the art tor example. by SEN In some emodiments the majo tund in Certa embodment, at least 60% 70%. Q' 905 9% 99% 15%, M9 Y, 990 9 O 100 r btn O m0 % to i0%, MY to 7% 6N to 75%. 754, to 0' 754, '? to 90%. SOJ . to 9.) 0 to 9' k 0or 95% to 9 ') oW thY x\G na filown, are between Ito 15, 2 5, o 14, i to 12 ro o 40, 2 to 10, 3 to 1 1 %o 1, Ito 9<2 to 3 to 9.1 to 8,2 to 8,3 to 8, 4 to 1 to 7 to 7.3 to 7. 4 to 7, i to If to f, to 4. or 1 to 3 microns in lentah as measured h any method known ato one skilled in the art for example, by SEM 10671 In a speeoifceoa diment the majoity (atnd ian retain emlodiments, at lat 0sO 70 80% 00 W" 9 4% 99%, 99 V% 90 8 99 9', %or 100% r hIt w b en 55% to 6591. 5 t75,5' 65% to 85% 755 t to 90 80"% to 95% 0" a to 95%, or 95 to l994 ot the. sNA j runoiers are about 8 A Q 5, 1. 3 or 2 microns o ibh ais measured by any method known to ng alN.died in the a, ir lannle, hr SIM. II mother peclil e mbodimllent, the maiory vamd in aeaim mbodints at least 60% 70 50 C 00% 89 A 915% 99.8%, >N 99'< or 100" or bytwen Ax0% to 65% AN% to 75%, 65% to 75%. 7,5% to 85A 75% to 9&i, 80 a 4 to mo%95"><, 95 or95% to ) o the sAG! nanoiThrs are between about ? to about 10 miern, about 3 to about 8 micrOn or about 4 to about 7 microns in legth. a masured by any method known to one kled in the art for eampa, by Sl'M In another specific emodincnt, al ( 00%) of the N AG nanohiers are between about 2 to about 10 iemns abom 3 to aboi 8 yrmns or about 4 to about tnicruns mle0ngth as ed by any method known to one s Vd 'm the art, ior exanpile, byi f 0068 in tn embodi s, the sNAG nanofibes fiberrein a rne between 0.003 to 5 mcrons ithekness andb diametera de trmined b electron rnmrseopxs hn speciitc embodiments the sNG nanoibers are about 0 0 0 0 0 00 00 06, 00 00 0 0 02 0 0 5 0 40405 05 06 0.65 0. O735 0.. 0.3 0 9 c 1 0 3. erag or yrang be.0.02to 2 rons0 t rs 002 to 075 l~rnk.002 to 0.5 rir1.00 o05 ndmicow 0.05 to tnviron, 0.05 t, 0 75 picrons 0.05" n h t in n .0%, a 91%, 70% A. 15%, 998% 99or 95%, be% e hi to6 % or 55% to t.%. 55% to 79, O% to 5% 709% o 95% 701 to 074t 05%, 90% o 95% Or 95% to 99%) of the MW nloiesao naner haes or diMnotw ot sho ao 02 to 1 deroh or o enibod0int ce mpajonk' tvnd in colo ai~d~ tit own 60 7>0' % SO, AM% 95t'A 94"?, AM% 9910", 9914 919% or- 0 SM. or between 55% to 10h 5" > to 7M. 611 to 7.50 791 to AM. 75 to 46% 5010 95% 9M~ 0 , as AM% or 950x to 99%) of th SN\AG u wtoiers Taxi a thicknsor n ctrot a nob 0.0i th5 niot hIn palic c odnnnt al t 1 0% MY of be S nanoibers have a ticknes or d2isetein ao 0 02 to 1 Kix0ns or bo 05 t o 0,5 microns. In ct n erbt a vens, the narta it n n et at Las 60 0as 60% 0% 90q> 0 9 a9, t 055oo 55 to 9k%, 65%1 to 75% 75%, to UK% '7514 to 9M,4 AM> 1,to9501,9AM to 95%. r9M to 99%) of the sNAG nanons have a thnesWs or dia e toonnt 0 2 tho 2 haon 0.a to ks m on 0.02 tof 05 krons, 002 to 0.5 km ss 62u5 5 Domons 0.5 o 1 oy 06 to 0k Do 00f -Vo 0,5 mirn, 0 1 to 1 nion.. a 3 to 05 Th ws.ro or 0. 1 to US5 nmos 35k9 30In cornaw enbodimets. the nunor'and in c certain embodime at 8e"' 90% 90408', 9M 995, 09 ',> 919" or 100M, or betwan M to 65, 5% 75. 6% to 71 n' to 85%s, 75^ to 90 0% to 95% 9.. to . 'A or 950 to VV't o 95 sNAG% ma)ootnvt V to ce% ita\ h U microns in lend adha a thikn or03 di 0e D 10070 Kn certain enthKodivns. th nroK'cv'x> wezht of the sNAG nrofl xr s v .'!stha iO5kDa, ~kM. M 0Da, 760 D 7 kDa 6k 45 . D 4kL< 33kiDa. R5 DK "r Kk kDh l 35kDa. S0kDa. or 2SkDa. in. oerain. Wnord tnams tAe fnorix (and in crmrair crnbodiZenni at least 60% 0 80% I 9VA, 9M'AO"', 99 19'~0 8%. 99,9Q, or '100VA orb Moaen 55% to6 .,55% to 75. 69% to 75M~ 75> to 151b, 75W to M0l, F02 to 9.5%. 90% to 954k or 954% to 49) ol th AG no nnovtir bwnn,.v%,a -iio~h wn<Ice ivvg of Less than 1{IOKLb. M4ktV' Wkfa, '7VDa '0 U nK u Gkia Tk VMS, 4RE :13 MA W0k., 39> 04so other embodiments the innty (and certain enmbodi ments I at least 60% 70%. 80 90% 95 8% 99% 99 8' 99 .8%, 99 ' or I 002' or byetweenr 5 to0 ... . 35% to F 65%4 to 99,9110 75% 7% to 85% 75t 9%,80 to 95%, 90% to 95%, or: 95% to 99%) of the s'NAGJ nanofibers have a molecular weight between about 5kDa to 100kLDa, about 10OkDa to I100kDa, about 20k1)a to 100kDa, about 10k14a to 80kDa, about 20kDa to 80kDa, 2Ok1)a to 75kDa, about 25kDa to about 75 kDa. about 30kDa to about 80kDa, about 30kDa to about 75kDas about 40kda to about 80kL~a, about 4OkDato abou 75kfaY about 40kDa to about 70kDa. about Sok4a to about 70kDa., or about 5S5kDa to about 65kDa. in one embodiment, the majority (and in certain enbodlimenth, at least 60%, 70%, 80O%, 90%, 95%, 98%, 99%, 99.5%. 99,8%, 99.9%, or10% or between t5% to 65%, s5% to 75%, 65%- to 75%. 75% to 85%. 75% to 90%,. 80% to 95%, 90% tos 9.5%, or 95% to 99%) otflhe sNAG uanofibers have a moleeular weight o about 60kfa, of4 althe s\'AG nanothetr are deaeetv -ted I sonme embody ntr c % O'0 mta 2) " o 30'% of the -N Ar nanohiber> are deacethard. in other embodmtms les thm 304) 500 N0I%, 15%. 0, 0$ 5Ve . B, t,%'' or l'eofthe sNAG rlan'fibers are deacetylated, i some imbodimemt eg aa to or mnore than 1-3 5%, 10% 1% 20% 5% 30%0 25% 40% 4-9, 500Ly, 55% 6s n3&~9< 70% 7'49 804,< 99%, \)\*v, o5" 9i0% or al I 100"'i) of the $\G nnothers are deetv lated tn other embudnv-'m.\ le than 1% % l0% ,5"o, 20 % 2Th% 30% 35%, 40"< 4-St t' s 5% 60"% M 00'e "S> 7, N0->, 85"-o, 40 05 i'9% or 100% of the sNAG naiotibers are deneety latedil 1100-21 in crnain embodiments, ~0'% to 80%, ~7"% to 8)%e, 7"A to 85%e, 8.5% to 95"%, W9 % to 95% 0%4 to 99%3 .r 3K" to 1 00"% of the sNAG nanoilbers are acetvated. tn sone enbodinients '0% O'75% 80% 85"% 000 ", 95% 9%, 99% or 100%, of the sNAG nanoiThers are acety tated bV othet e.mbodi 001ts moura tid ~ 0 U3 > 75, 80%, 85% 90%. 3, 59%, Q9% o 99 5% o0 9% ot-' <NAG nanofibers are acetylated, In somec embodiments~ equal to or more thdn 12-; 5% . 10";., 15" >%5 ~ o 5 0, 4 -50 ., X-."o. 0ss 65A NI 7 80% 8 "- 0B 95> o 99K% or all (1 100%v ot the a AG (-nanofibersare ace-tylated, in otlier embhodimnto less ihauni lu 9, 1 1, 0O' 20, >% 30Y .5.40 .5"% 50% 33%, nO%, 65% -7 75N% 80%~ <.>.0 -'% 99 or 14 ot'th '\,\G nartonucer, are awCeyated 00731 ir some embodiments the s&\GA nsnohkibeconmn U at leat one glucosamune modosacch trde, utd mdv t'inthcr compi se at 10ist 10"-- m0s sl0') 40% 0 50% 60% 70%.o 80%.a 90% 95% or 494 of the NMeetylglueosanmine roinosaehbarides in other embodime"nts the sNAG nanofbbers cotmpdse at last one N acetyig.uosamine monosachatide, and may k10 h" comprise at least 10% , 30% 40% 50 0%, 0 0% 0%9$% or 99% of gl ueosamitte monosaebhan des. 10741 in one aspect, the S AG nanoibers mcrease the meubohe nae of scrunred human umbibeal cord vein endothehal cells 'C") i a \T asay A \T'l assay a laboratory test and anstandard colorimetrie assay tan aay which measures changesmn colors for measurmy Mceular prolferatuon cell grow tht Bret fellow \N Y ' 3 (4iimeinvlhiaoV> ylb2'5Idphenwlctrawiiur bromide, a tetraole) is reduced to purple formaan in the m.itoelmdna ot hng cells. Tbhis reduction akes place o&y w hen mutochondnal reductase 'enr<mes are acrve and therefore comversiont can bv directly rlawd to the utmber of vifable (la ine cels. The metabohi rate of cella may be deteainnd by other tehnques commonly k no to theskile d arisan 0751 in another aspect, the sAA namotibers do not r rescue apoptosis ot serumatarsed RC n atry pan blue exclusion test. A trypan Nue exclusion test is a dye exclusion teat used to determine the number o oable cls preset in a cell suspension is sed on the principle tha lve ells possess iact cell menlmanes that exclude certain dy es such as trypan bhie, LoAm, or propidium, whereas doad eccls do ut, The ilit f cns may' be determine by other techniques commonly known ho the illed arm m 1§D761 in certam emb&damnts compomions copnsixmn the sNAG nanaof[bets me described, hereimn the sNAG nanoStbr increase the metabolic rate of serumrvaned human uumhial cord vi e dothebal .els in a vItT away and/or do not rescue apopiosis of serum starved human umbihial cord vei endothchal cells n : trypan blue exclusion test, . some embodipments the s G nanoibers ierease the meibohle rate of serumsian ed human umbical. cord vein endothelial cell i a \IT asty and do not rese apoptosis ofsrum, statred human umbilcal codl ee m a wrypan blue exclusion test SU0771 it a specific embodinmem the WAG nanofibers are biocompatibe Biocompatibility muy be determined by a variety of techrtques, nu but not limt to uh as the elhion test, itramnusehar implantatons tatan or syUeac iniectitr imno animal subjects. Such tests air described in S. Pament No. 6.6%342 1sec, ag F-smplc 10), which is incorporated bv reference herein in its entirety.

00}QIl In certain etbodiments, the sNA \.tianoriberti sed in the methods described helein are non-reactuve in a bioconmatibilit test or tests. For example, the sNA\G nanofibers used in the methods desenibed herein may be nom-renemev when rested in an eta tum test an intramluscular umpiantatma test an Intsa.Cutanleua test, and or a systemic test, in othee' emboimens the sNAG nanofibers used in the meThods described herein hav e Grade 0 or Grade Test score w hen tested ini an elutton test, an intrainuscul at a mp imtion test, an intracutaneous test, or a systemte test hn vet another embodiment, he O\ tW ina others used in the methods desen~bedi herein arc at most. mildl i)ittatv1 nwhen tested 10 d e.tto~n test, an 0anseulaf ilnplanidtBon testa iiTactttdneotis test, and/or a1 sy sternil et\ 'd I certin1 entbodlinents the copositions described herein do not cause an ullergrtcni reaction or an irritation. In other embodime~nts the eompositlons described herein cause at tnost anild allergemec reaction or a iltd irtioln, eM. at t1he site of application. Thle rele\an 3 tests anid evadhuation of test rotbilt are described in, e p, U S' Patent No (N436P hich is ineorporated herein by refteoe 'in its enflreiy and in Sceuon 6k 5,a2N 90 n a specrtic e'mhedime nt, the. CN\AG nanofIbhers are nonseactrvn> whenlt ested in an imtramiutenhr 'mplanainn test hi one aspect, an minrainustnar .tnpluanaona test is an intramuhenbar 'rmplant an test ISO 4 's eeA iplantatcvn, as deserthed it 'section &%, in/ta. In certain emnbodimenms the sNAG aotfibers display no biodogical reactivity as determined by an elhibon test (Fluu(on Test (lrade 0). In sonec etmbodrmentrs the sNG nanofihers hav e a test score equal to "0" and/or are at most a negicgibie irritant a~s determined by itacutancous iniettest 0. in SOme Omeemoom n s the sNAG nantIbers elii no itrdermad reaction (cc . Grade Ireaction in Ki imnan test and/or ha\ve a wecak allergenic potential as determined by vh Uaap. test [0Q80 1 certan aspeo. thbe sNAG nainotiner ae itmmunoneturai(e.. teydo not elcit an immune response 1 nanoitibe nprefer afy degrad" wuthmr about I d~a 42 diay i day 3 days d'cay s w..ek), 8 day%,0 Ida mi '12 days. 14 d as C weeLs, das . I daya 'u o rek. 2: day' 3 da'r (4 weeks) Al dc\ 4, * 1..l1 I days, 40 days 2, ci' 59 30 days >$ dd) 60 add kv months 63 days 70l days 75 days 801 days 83 days 90 days3namembs. 95 days 100 days or 4 months after adn rat ion or "lion t a atadt 2b OWN Z " N 100821 in cetan ebodiments the s\AG nanofibers do not cause a det ectable ieip body reac-ion. \ reign body reacuonwhich may occur durM found healung, includes aceumuainof of exudate at the Wie of uinjrv infiltration o'nflanimahrs ce bo debriti the area, wnid the formation 1 gla t don tissue he persient pr essence a' 'o rgn body can inhibit fid healing. Rather than the resorption and reconstuirm that occurs in wotnd healin, the foreign body reaction is eharaitenzcd hW the formation of 'orexgn body ant cell.A encapuILation o 'the orcign obect, and chronic int'lamYation hneapsudatou refers to the Varm, generally av acular collagen shell deposited around a foreign body, eteetivly isolating it trrm the host nsues. in one embodiment, treatme n of a ite tg, a wound or a sie of a bacterial mfection i a wound) n h the W \G nanotibre doe not elcit a detectable foreign body rea o in i day, . dy 4 days 10 da r14 y o 1 dav ater treatment, in one such embodiment, treatment oF a sie Q , a wound) with the AlM nanmihes does not elicit a foreign body encatalatr in i dn 3 days. 5 days, ' das 10 dayso or 4 days er treatment 3 In some embodiments, the A nanotibers (a omn<e liners, wern mar "t of the: fibers are betw een about 1 and 1 5 mirons in length, and (ii) lincrease the metahohe rate of somsatred EC in a MTT asy andor do not rescue apoposts of scrumstarveA P i a trypan lhe exClsion teb ad (b) are non-reactive when tetcd in an intramuseniur tntplantation test. in certam cmbodinents the sAG nanofibers ti comprive tbers wherein mortv of the fibes are between about I and I nAcrons in leN and 6ia hnrease the metabohe rate ot serumvstaved C in a M assy and/or do not rescue apoptosis ofsermvstarved£0in a trypan ue exclusion esnd (b)new uomacive vMen in antntrn uiardimplanta test. hi cer taianbodmrs he KNAG nanoflibers omprsie iers, herei na'o'ty of the fibersare beetaou 4 and 7 m ts in Wgt and id}in rease themtbob ate o sommstarved ECin a MTIT assay and/or do not rescue aptosi of 1Jrnstarve tin a rypan blue exclusion test 'n)d e nonreactive when tested in an ritramuseCd ar inplantation 00184j In certain, erbodirments the sNAG nanofibers do not have a direct efitet on the groth r urmalof batr& Saab auni, awren as detunld byon siled m . th atlote embodhenms sNAG nanofibers do not hav a direct effet on the growth or sUrvival of bacteria such as S, wre as determined by the methods set forth in SeCtOn A4'25. infrw in some embodiments the %A ( na&noih ers do not have a incet e et in itro on baetenai grown or sur'o al. In one embodiuent. the sNAG nanoibers io not have a direeTect e g , iii oir) on tro th or sur\ val of r:wnega~tw b bacteria in anotherr nbodinMent the N AG nanolidbers do not hav e a direct ell'ect e g m thro) on gron th or s Ud i a r ginpositive bacteria. In vet another 'nbodimeat. the sNAG nanolhrnIs do not have a direct effNt t n W vt) on grnth or survival of either arpo-pine or gra-negiatve bacteria in some ambodimeiS. the sNAG nanotIhr or a s WA nanofIber composition does no bind bacteria (e'g., graVosUNe bactrOa, gtramnnegativ bacteria, or both types of bacteria) in NsC embodies, incubanon of a baea al halture with the s% nanotibers 1e gc , 5000 pag of s\ iiauters) mn itrO does not redue bacterial toad K 10 tmants, 30 hinut "s hour 2 hgrs, 3 hours, 6 hours 2 hours, 24 A00N. 4 hourN. 72 hoursl ii 9 2 ou ocuhation (wherein the bahaciwa I culture Cd) be gram-posiuve and or a graimnnegAe In some embodimins, incubate of a S aiu culture sith NNAG nanotibers (g , aboin Xdugfdbpg, or about 1000{0 ag of A\KG naoiber) m On does not red ue baceri load in 2 hours 3 hours, 6 hours or .i hours oincubation. in yvet other embodimenit the NAG nanoiNers ieaduce baetel. grow t or survival in VUO by les than I log, 0 r loX 0 00 log. 0 log. .4 lo. 3 ' . 049 03 on. 0 5 log, 02 1g OJi 3og> 0.05 lou or 0,00 lo tor example, when Sqrini m anans bacteria cehures are treated mubated oith the A\ AG navifers Gi vitro In some embodiments, the> s\ Mi nanotibers reduce baterial growth or surit al in vitro by less thon 1 x 10, 2 & 10' 3 N 10'Xlx 1 0 5 x 10 6 x 1A 7 x 1 Al i ;0. 10, q 10 or 10 x 10' fuml for example, when Staphl~1ac/ m anreus bacterna cultures arc treated/incubated with the sNAG nanofibers in v\ n 'ht* itests of'the effect of sN\G nialiThers on bacterial unirw orvivand til ev valuation of the tt results are described Or examnle,; int Lne 2c 2 . Section 6 2,5) and 0081j hom embodnmentls sNhAt nanoftbers O epielbrWlri aol)i the fibers arc between about I and 15 microns 1 and 12 microns or 4 and ^n mlrons m lengih (ilw do not hawe an cffc on bacterial grow th or survival of saph inocOcanu wn n bacterial cultures in wiAr, and (ii) are nonl active when tested in a hiocompatibility st (eg. an invraniuc hidr mpelntation te't) f00861 h certain emMbodin e A( nanotbers idue a e rtain pattern oj gene eopressRon (N or proteinxas detrmined be RT PCR ninarray or E SA) in a cel tssue or organ treated with or exposed to aNs4(inanobber coromptdioan Spec ifically in some embodiments, the sNAG nanofiers or a compostion comprising the sNAG nanofibers Induce expression of one or more defensin proteins one or more defensin-like proteins and/or one or more Tollike receptors. In yet other embodmems, the sNAG nanofibers or a composition comprising the sNAG nanotibers induce expression of One or more piroteins that are known~~~~~~ to, haeaanttceiaeihr [00871 In certain embodiments. the sNAG nanotibers or a composition conprising the NAG nanofibers induce expression of one or more odefensin eg., DEFAI(i.ea-defenin I). DEFA IB, DEFA3, DEA ,DEFAS, DEFA6), one or ome p-detbnains (e g., )EFBI (ice. p defensin i ) DEFB2, DEFB4 DEF103 A, DEFB104A, DEFWWSE. DEF3107B1.DEF10OB, D.LFI 10, DEFBI 12, DEFBi 14. DEFBI 18, DEFBi 19 DEFB123, DEFB12. DEFB125, DEFB . DEFB 127, DEFB12, DEFB129, DEFBI3L DEFB136) and/or one or more 0 defensins (cg DEFT ! n some embodimems, the sNAG nanoibers or a composton coprising the sN AG nanotibers induce expression of one or more of )DEA 1 DEFA3, DEFA4 DIEFA5, DEFBD, DEF I bDEFB103A, DEFBI104A, DEFB1 I08B1, DFEF1112. DEFB31 4, DLFI 1l8 DEFBI) 9, DFBi 23. DEEFB124, DLFBi 25. DEFB...26, DEF12 D)EFB29 and DEED 131. in certain erobodimems the AG ranofibers Or a compOsition comrising the sN AG nanofibers nduc expression of one or mor Toll receptors (e.g T LR 1, TLR2, T1 U R3 TLR4, TLhR T LR7, T5L.R TULR9 TLR{10, TLUG I and/or TJ, L ), in otter h NAG nanoibrs or a composition comy iig the sN AG nanobors induce depression of one or more of IL-L CEACAM3, SPAGI % SfG P (L -ike receptor, 1RAK L IR AK2, iRAK4, TBK 1, TAF6 aRi IkKi. in some e.mbodiment the sNAC nanofibers or a composition comparing the sNAG nanofibers induce xnremon of one or more of IRAK2, SI IRR, TL I R" TR4. bLR7 TLRt TL 0 and TRAIF in ore embodiment, the sNAG nanoters or a comn osion eonpritn the sNAG nanofibers induce expression, of at lea one oFthe abmV-lit d gene product, I00al in)m enUbodienth NAG nano0ibrs or a composition Comprisuog The sNAG unnoibers induce expreion of one or mor e or hcie -ised genes in the amount equal to or more thani about 0 2$ told. 5 DM6. . Pld, 1IW fok 2 fold, WK5 d3 16, 35 il, 4 fold, 4,5 fold. 5 fold, V fil, 7 foil N d. 9 0 d fold. 12 fold, 15 fold or 20 $o aN corned to the leve of presin of the one or more of the above-hed genes m a celtisse or organ of a subject before reatment with the KAG nanotibers (eg, a known. aierales Iel of epresion of te one or inore of the abovehted genes). in some. embodiments. the A A 0 nanoiers i a conpOsithon Corn)pr~iOi the sXG ninotIbers induce eptavyl of one or more oft ho Atov Uted genes pi amount equa to or more than Mhout 1 1 5 0% 75", 100%, 123 ' 1VON 1'5% '00I 22' 250%4 275% 300%. 350% 400% 40% 500%. 550% W00% 60%. (0l, 750" N X6K ,000% or 1(00% the level of xpression of'tO one or more of te abvei-hted genes i a cell tissue or organ of a subject heforetreatrent with the sNA\ nanofihers (et a known average let el of expresion of the one or more of the aboVe-lised genes u 100891 in Some embodieis, the NAGj nanotihers but no iong naP&eetylIus t mme ehitin and/br lhosan mduce expression of the one or nor y listed above, as determied by a method known to one skilled in the art, or described herinI Nloe of these embodinentsh ong poly. eetyiglueosanme, chain and/or chimtosa do not induce epipession of te one m iore aenes lited abo ve or induce lower level (e gn, more tan I > fold, 5 Iold, 2 id, 2.5 told. 3 d, L Adt 4 bld. 4 fbdk , i fol, 7 fld, 8 Abid, ) Iidt or 10 Iid lonwll of expression of the one or rea veanes ited above as compared to the Ie\evl of eniession of the one or more genes hsted aboe mduced by the sVAG nanotlets as deternued by a method known to ona shilled in the art. or described herem,. 00901 In certain cmbodiments rho N A G nano tbers or a opposition comprising the sNAG nanofbet s iduce a nene exresson profile that is consisem wh, similar to, about the same at or equiv ent to one or more sene xpresstOr proxies demonsilt~d i ldbes 1, iL hi, V and L\, Setions 6 ~61 ir a In some embo&unens, th s.jAG namofibers or a composion upregulated by sN AG reatmem in Taies 1, it, 1l V, Vli and I\M Stonrs 6 NO, 0r, h some embodimns the SNAG natihers or a composan comipring the NAG nanolihers induce expresion of the Mnaiority r all of tC genes shown to ne uruutd by .NAG treatment mF Tables L1 L V.1 V, Yli and MX S0eUo n 5 ito. i some of MAie embodunents gene expression evels ire measured at 1 hour, 2 houirs4 hours 5 hours.> 6hour s hours 10 hours. 12 hours 14 hours. 16 ours. iS hours 20 hours. 24 hours. 48 hor 3 days or 5 days after treatment at a coil ti\kur or organ w. ith a ClAN anofiber composItion by a method known to one skilHed i the art, or described herein. 26 100910 in ceram mbdiments the s\ \nanotibers or a coin pos ucomplning tue s\\G nanfIbers iiduce a gene expression profiIc that differs fToM the profile induced lw l O' pol\ -, acergtucosammne fpolvners or ibers in specite embohmemsv a gine im proli induced by the sNAG osnofibers is consistent ih, sirmiar to, ubout the sane a5 01 e.t un akem to dIat show n Tables 1,H, l . , V H and1 IX.Nections r, 4 ' nrX where gene expression prof I indu cd b) long polNa-etvIgiucosdA noe polyers or fibers is mnswnt with, simdar to, about the same with, or etiAuiKi to that shown in Table VA and or L\ Section 6,, nye In other emboduirt ~.. the \ nanotibers or a comipositm compriun; the sAtA nanioibers 100 tgene e*\prtbsion p~tl lhICl e r n toe. gene expresssin prTlde tfidtltWd hr ehitin orchi tosari 10&O92J in. ao spcifi embodime'--nt the.S nawofhers are obtuad by irrdiating ply-N~ acti ue os l m ae a nd r ad e m v ati ne thereo f. S ee ee ti U5nr g d g p acetviglucovlmne and denarixc thereof and Secton S 2, fi regarding methods ror produ mnn the s\ \G nnotfars ua g in'adiaton irradion iay be used o reduce Ih tn of pol Y acet wnlucosamne fies andior poly Nuoetylglueosamme dernvatne fibers to fom shoriteneId pel )~&3II ac~aezvkgliotamxe liber's<anctor shortened poey)~taeylglioanubamle. deri atn P ties is NAG i anfibers, Specirleafly, irradiatkin may be used to reduce the length and molecular weight of poiyfly ac~r)lucosanine or a derivative thereof without tbtbm itus mtcrosctiue. The infrared sWeetrn dIR o l'AG natnofibers is smal tA the same as, or equivalent to that of the nw-adiated poly-f, . -acetyhruleosamme or a den vise the ireof. f00931 in one embodiment, the s\AG nanotbers are not derind Aomn ehitin or Chitosan, Iheres in another embodirnet, the compositions described hbrem may he deiled or ehitn or cotsan. 0r the SIN AG nanofIbersna\ be de 'ed from chitin or ehitosani iA I Pol Acetylgiuccsamnine and DOriativs Thereof |0094j U.S. Patent Nos x622t34; .62106; 5ji~4679; 3,68a 1 13; 5%350,AS; 6,59i720: 6060A342 1 5 30 and t .S. Paren Pub. 2009 U7 ~75 (each of which is incorporated herein by reterenee) describe the poty-N acetylglucosamine and derivatives thereof and methods of producing the same, n sone embodiments, the poly\,aeetylglucosanine has a f-4.-4 .orldturatiOn. In other embodiment, the poly zegtyineosarnine has a n -4 tmigration. 27,i Thepd o deacsaanand devat ie heof may be in h frin of a polmer or in the form of a fiber, 0951 Po acetydiieosarine can. for example, be produced by and may be pified from, imrala. preferably diatons. lhe atoms which n be used as staring sources tor die producttm of the poly acetylucosumm( melue, but are not muted to members of the Goscincu gms, the (iw'brc'/i " enu' andm tiae f~liasms genus Pety\W cet~yhlucosamine ntam be obtind iom d'tom cuturen in a numher of different methods meluhnm the m.nechanteal Tree ctliod and trmca bt'ioloteal method loon the art (see, r>Q. .S Pam rNos, 3/22,34. 5,n: 064; " 24/6?9 3/86, 13; 't Z3% 6,5W7NI, 6&86,342: and 7115i 8 eNeh of wn hih i incorporaed herein by reteence in its entretyr in certain embodiments the polyNa etygueotme is not derived rm one or more of the ~lovr a shell fish. a crustacean ans!et a nogf Or yeasts [0096 i one embodiment Poly 1 4ac"'yducosamineis derived fron a process compr g iraa treatng a algecomprising a c. body and a poy N cetgiluebesamme poviner Pher with a bIological agent (such as hydrotlurnei capable of se pirating thc N-aCewll yucossmne poly mer fiber fron the cell hod, fir isufteentime so00 Oht the polt - -1 N 'is lgducosamme polymer fiber i released from 'eli body, b) segregIatr.o the pol- K 1. .t4Ncety l inem polynr fiber from the cdM body; and et remrox'rug contotmnants Worn the segregated pl3 U -. -4 Fuetyiglucesnamiw polymer fiber, so that the pod- o1 a miacetvhglucowmne poinmer is isoled ad purified. 0093 In other embodimtentt the polV / ,-1 N acet\Iglucosune may be deriveCd front one or more of the fSolowng a 4h>1l fish, a crustacean an im rYt, a fung. or yeas in erIn emtodimenzs, the compositionts described herein do not eompmv ehiiti i rehitoad. [00| One or more of the monosacharlide its of the pox- A-acei) acesainmay be deaccylated in certain embodiments. 1% to 5", 5% to 10 A o O 20% to 30% or 20% to 30% of tie poly cetylghtcosamine is deceylated In some e I % 4 , embodiments less tha 30% 2% 20% 1 10% , N3 2 or of the poly aetvyglu we ine is deacetyated. In some embodiment equal to or inore than 10% 11 1 ~ 0%h 25W, 10%{ 5~ 3 . 0% 45%t 50%, 505k4 60%+ 654+. 701% 7394. NO1 45% 90%, 18 95 or 93,o ai (1 004,o h poKN-cewydtCosaRin ~deacoated i oae Cmbod itrntS. eS taos %1 , 0 2 5% TV 0% 402 254 3 %t A% 55% .0%. 65A. 70%. M% NOVt 4534 03.5.9%or00ofte WN eyuosai s deacetvatd. 1099 in cerain e odiment a p N weg sam compoSio tn comprisea 701, to 804, iS to 80A5Mt o N 8%5" to 95% t0 o to 5%. oTV at M or 9M to 100", of actisthktted glucosammne (ie. N ,\ety ucosamine) mhonosselaiddes lin sme enmbodiments a poly- ncetyglucosdime COtypOttim comprise 0, 754, 0", 854, 00%, 95, 94, 99% or 100" o' aceO y ated gle sanine e. %eety teosamine t m0nosaCChades in other enhodirneutt a poAdaetv ig|Ucannne COo~fpoo L00 amp.ids more than 70% 75 V4% $5 90, M 9k 90"4 , 99 t w or 991'% of th acen lated ylconsamne in some emoient a pcI ~Y-'eetltjccosarnine e01wp03lt lo conpis es Couan to (u t C tntan l 's 5%, 100 KA M IM% 734, 5% !V, 4 4n 0%, 55% 60% 650. 703S. 25%, 1 0 Ax 15%,) 90%, 9% or 99 A I ni a (100'T%) of the awCtylated pluCOSitIfle i Uothet er1bodrnents. a 1oi c -aeet lueuo i mosmm i tiOntff comprisO isS than 1% 4 , 10% 15% 20% 25%. 30%. 35%A 40%, 45% 504 5%, 6o 65%, 70, 75%, 40"4, 8 90> 5", or 100% of the 1001001 in %om omboditnt, a vle i eompotmon comiNses at least on> alcosarnn i0080hne an ma furhn com pr&: at teast -0% 20o % m 10. 40*> 't 50% 60 . 4 ,.0'. 0% 954 OT 99% of \f<crnlg osamme monosucehardos, in other embodimet, a pom- N eyilucosamme m comIpNton orpinses at least one N e~tylplucosmine mtonoaachartide, and may further eomprhs at lea t 10', 20% 30%. 4(0% 50%. 60%, 70 5% 904% 9" r 99' of gluesaline m1no9ocharldes 0 1(1 Drivat6vs of NO)~utyigalcosamrUe may also Ie used n a eot1'ao10tm described herein, Deniwi e of polyNaetyigiucosamme and wethoda of makmgsch dei .a t.:9 v dacihad m US Patent 53, . 23064 (see e w eti 4) nhich is eorporated b reference herein : its emtiret, Derivatix e5 tpolyvNactly eosamine may ineld bui ar not himted to, pantialy or comQletely doacerivled poly-cety iiuc samnUnr or it deaeetvlated dcrivaves further, poAy Sucety igiueosmme max bedem atiaed by bem sulfated, phosphoerxa ted and/or NiNrated Pol4vnacetyI o nP derivati es meluado, eg, sufAted poll Aseg ljohamine d a s phosphoivlated poly.:aetyvlucosamnine I0 dem vatuvcs or nitrated polyt aetylgiucoaamne dernvatmes ,\ddi timaly, one or more Of the nmonoaaccnaride unis of the polN ,ewrudaucosammne maw conmmi one or more 5tuh'onyi groups one or more O-saeyi grmups. in addition, one or more of the monosacchandes of the dear itylated polvdacesigiueosartnme ay contain an Nacy! group One or more of the monosaccharides of the polivNacety lglucosme or of its deacety late darin aive. may contam an ,aky I roup. One or mon of the monosaccharde unis of the polW A~naee ilucosamne may e an alkaIh derivaive. One or more of the monosccharide unis of the deacetsiated den vairve of poV>& acetviucosarmmne may contain an Nalky. aop. One or more ot the monosacebarie onrts of the deacetylaned derivative of poly~ -arty lyucosnmime may contan at eat one demyhalogen den ative. One or more of the monosaeehande unis ofthe de cctv ted derivatve oF poV N acetWIgucosamne may fAn a sat One or more of the monoccharde units of the deaetylated deriatiye of pol)Ai-aceety lluosatnine ina~yfonm a metal chelate i a specific embodiment, the metal is aine, One or more o the monosaccharide umts of the deacetyted de iv atie of poly~Naeetyl yuosamiie may contain an ~il kyaifue or au NtaryhdeIe group, in one embodiment. the deratWe is an acetate demv'ati. ;,n anher embodiment, the derive aft\e is not an acetate dernhvir a on" 'mIoodiment the polyN uaet11igl ucosainxne or deacety lated wo )~aeertvIhensamknn i deri atiar wdi lt Iaeu meaid. Wherein, in another embodimem, the derivaie is not dersvmaed Wth lAtic acid 100102| 'Tho poly Nacty iflucosamine polymers or fibers, and any derivati vs of polytN aceti ;lucosamiuno polymers or fibers descneed abso e an he irradiated as dry polymers or 1iherS or polymer or tiber membranes. Aternalir el polv acetytlglucosarmne polymers or fibers, and any drivatvae of poly acetlyigimoamnc poiymers or fibes describd above, can be irradiated w hen vet fhe methods of matig, . nanofibers by irradiation and the shN\G nanotibers so produced have been described in 1 Patet Pub, No. 200900 7173 which 1i incorporated by refer'ence herein in its cemirew.y 1001031 in Conaik embodiments the pol -N'acetYhgucosamime po rnets or fibers are formulated into a suspension'siurry or wet cake for radiation. Irradiation can be performed n ... nctly wth or followimg the vrmulation m the poly.er fihers io ats tina formulation such as a dressing Genendlv, the polymer or fiber content of suspensions/slurries and wet akes can vary, ior example frm about 0, 5nm to abou 0 mg o polymr or fiber per 1 30 irts or disrfled vater are used for slurries and from about 30 mp to about I 000 mnt of p ner or iber per Pi ot'dimilled nate are use m we t cake formulanons. The poly mer or fiber I st he yophi d, froeun in liquid nitrogen, and piuverized, to make it moure susetttle to fornintasispensamnsbirr or wet cake, Also, the saspeosams/skuries can be theired to r emo\ e water sch, that a wet cake is rOrmed, in wernam aspects the polymer or fiber is irradited as a usenSiOn compsmA aOnu 0 5 mg, I og. 2 m, 3 ng 4 oy. 5 ig mg. m g a8 g 1 0 mo, 12 mg, I mg, in mg, o 0 mg, '5 my or 50 fpolyme or fiber per ml of distilled water, or an6 rnre ci bet\ 'o die ioing emdlments W>' 1-10 mg mi W5 g NLA m-s nml 20-50 ng 1ml, (s ) In oilher apects, te polyineror file is rtMdWited as a wet cake, toinpiSAiCg about 50-1 A00 mg polyme or Aier per I mi f ditilied water In speeitle embodiments. the wet cae comprises abou 50, 100 200 300, 400, 500j 600, 700, 800, 900 or 1000 ni of po ymer or iber per I n Y Jitlled water, or any range m Atween ( g , 1 0000 mglml, 300,600 mg mL50 1000 m g Am. 0tH104| lhe radiationn is preferabl, in the fon of'gayrna radiNaum, c-beam riation, or rays Two souros ot irradtioin an Prfcired radtoactnve nuchdes and elciricity, in speci embodsient. the radioactive nuelides arc Mb hAWt and cesium- 3 Both of thee nuchde emit ganma tiy which are photos contmng no ma The gamma n ray has e energies Rm 066 to l .3 Me ,sing electric ty, electrons arc gcnerated and aCeclirted to energies up to I) Me0 or igber. \\hen irradating polymers or Pibers 10 reduce their s ae colrsidernton to t ake into account is that the depth of penetration of materials with desties simiar to water by 10 MeV electrons is listed to about 3' em with one sded exposure or .about 8t6 em s ish tw oaided exposure. Depth of peneutation Jreases at lower electron. irgc .e aIO eigs can he convertd to s-rays by pl1ciei' a n"'mal (usually tungsten or tantalum target in the evecto- beam path Conv version to -rays a lmted to electrorns with energim up to 5 \ Xry are photons wih nomass and can penem polymers or fibers similar to ganmna ras Onere s only about 8% efsencvni. the Conreran of etron energy to ray engy g pweed eletIrn, b macinesare needed i x-ray production liesto account for tie lowconversionefficien 16 T orhed doe of dtn t he energy absorbed per uni weight of product measured in -ray (gy) or kilo gray kpy For dried polymers or hes te prtred absored 31 dose is about 00A00l kgy of radiatum, most pt.eferabl abouttt l'Si 0 hgs or about 00 U00 kgy of radiation, F or let poly mers or t'ierhe preferred absorbed dose isabout 104-500 \g of tradition, most pre ea b y about I0 -250 g\ or about N 0 kg of radimon.n o74 The dose of radto m . Jcr thed m tn ots efe on the length of the polymers or iers. In specific embodiments the doe of radianon u=ed preferably reduces the length of the fr vlmer or tihr by anywhere trom abonti On to 909 ol the sr gt of the polymer or jibM, respeev ely. in speciic einbodnents the aterage ength s Ceucco by about 110%. by about '0%b by ate 3 by about 40% by about 50% by aboit n0 by Maot 70%, by about 804 or by about 9%, or any range in between tse. 2010% 00 :N and so on and so iNoh \ liunatiely, We dose of iadia ion used preferab reduces the lemith of the poly mer or 'iber n to aw hee ton l to 100 mierons in specific embodimnots. and deptndm on the startog br leth, the 'avrage enqh of the polymer or fiber is reduced to is ahm about 1 microns ien than -bout 14 microns les than south 13 mum less than about i erons less than Maout I microns less than aboat 10 :mviroi less than about 'mtou ibh about 7 t1ero~nUA less than about S rmeron loss than about 4 incionns N than also'ni a n ons. less than 2 nicrons, or les than 1 Corons. In cean embodimonK Tv length of the njority (ard in certain mbodimems at least 60%, 70t 80% 90, 9% ", w4 915% 90 8% 19 9% or 100% or between 55% to 65%. 55% ' to 775% to 0%. 80% to 950, 0" to 93%, or 954 to 994% of the poiymers or flbors is reduced to no greater than about 20 aneron n ureater than about 15 ntcirown ri greater than nk ut il mileon, no grecater than about1l0 rmierons no geeter than about a n'rons no getter taJ About 7 micron or no greater than abot 5 alnero is cetamr nmbodmnts aa ton of thi pavcars or tileS iCOhi''the lenthi o the majority lad in certain nbod'mm. dt b it N0 80 909 . AN" 99% 9 M' 90 9 94 r. or I ,or between 55% to ON % to 70% 65% to 75%, 7 tO 15% 75% to 90% 80% to 95% ,904 to 95%1, or 9 Ao 's ot the fibrs to anv her" bew een about I to 20 microns. between abut I to ; microns, betw een abu to t ' nicrols, between about I to 12 microns between. about 2 to 12 microns, bawe about I to 10 mcron beMenc about 2 to 10 microns between about I to S microns between about 2 to 8 ronras btefntU about I to icrons between about 2 to 7 rmorons. between aboa a 3 t otront between about 4 to 7 tnmrons, between about 1 to 5 micron between about 2 to 5 microns 32a b tee about 3tonucrons between about 4 o 0 micrns, or ay iinge betee the foregog lgenhs, whichlarc aiso encompnassed. 190 8 O} he dose of radiation can also be described in terms oits effect on the molecular Weight of the polymer or tiber in specific hembodiments the doSe of radiation used pref Ny reduces the moleculu va eih of t puolyxcr 01 -iber by anywhere trom about 10% t < of the Sidlnng weight of the polymr or lier in speucWrnnduuents me as erdie moleculai wenht I radced by bout 10% 1 about 20% by about 30, by about 404 , about 50"% by abnout A K: abolit ~0% K abon 0 or bl about 00" i or an) rage n t bet en (eg , 20 dt0% 30 14'O and so on and so orT Alternatntly the dose of radiation uaed rrefcrablv reduces the molteut weight of'be polymer or fber to anyw here from 1,000 to 1,00000 dlons in specific, ermbodiments and depending on the starene, noleular wevht, the' ans 4ag molecular eight o the polyme Or r Cher i reduced to less tha n 1 A00000 dahkon o th an ~50, 000 daitoas less than 500,000 daltos less than 300, 000 o sdaon les than 10000 dahon less hanu 90, 000 dahons les han 4000 daltons, less than ~0,000 Vs than 6,000 dahos lews than 50,000 datons, less than 2 000 daltns. less than 10.000 duions or les oh i000 dtons In certain embodimenls tcm c eage molecular weight is reduced to no less tun 50 dalion, no less than 1000 daions no lew than 2,000 daons, no tens 3,30 daltons no t than 5000 daitons no less than 7500 dltons no less than 0,000 daitons no less than " 00 dadtoans no les than 0000 daton no les than n0, 000 dahons or Ro ess than I1 00,000 daltoIN Any ranges beln tzn the Ioi Poiwg avrag'C moleular weights ar e also enconmpesked, for eamle i cert n embodinmenn irradiation of the polymer or fiber reduces the arage molecular weught to anywhewr between 10,000 to 100.100 datons between 00 and 25000 daltons, between 50,000 and S00A000 dalns between 25000 and 100,000 dahons, 30,000 and 90.000 dahons. 1U4and 80,000 aons, between ahmao' 25000 and '5000 datons. between about 50000 und 70,000 daltons or betw een about 3500 and 63 00 d hns and so on and so frth. in ceain embodiment. mad ivin oftthe oh rne' or fibets reduces the molecular weight ofthe mnaoriy and m certain emnodinents, at l 601 70% 80% 90% 9% 00 99W, 991 in % or W00. or bwceen 55% to 05 55''% to 75%, 0d% to 75% '5', to 85%. 75% to 90%. 8,0%e to 95ea 00 to o5% or 95% to tl%) o te fibers to anywhere between about 20,000 and j00,000 datons about 25000 and 75.000 daltons about 30000 and 90,000 datns, about 40,000 and 80.000 d:50ns about ,00 33,.' aid NOO dAhon oraboe OO d 65,0 ridato n cerAa St,0odnnents Wd htion of he2 pOmers or bers roe e molecular weight of the mozrv nd in certain enhodimets, at '-t7t -60% 7 % n 90495Q k AN 99% 99 9 % 9 . or - 0%T r betwe 55% to 65% 55 t o 75W. 65 to 75O ON5 to 5%. 75% to 91% 80% to 15%, 90% to 95%. or 95%t99% o e t abou akon f001091 Fo!low.ingradiation Tarries can bet tered andded adet makes an be dd, to form conpostions c. dre es and oteretoo aitions describheein) that are uce ithe practe te inntn 53is C q5G Nanoi 10Pf101 ITe s'NAGD nano her ma tb ... mU-Oted in a Way~t of cmoiin o oia a~ Elm a qde scwo a mToww a det 00%O admin iaionn as described herein (ni I uj A composion comprising the ?n AG nanofers may he tormulated as a cream, a oemrane, a film, a liquid aoin, a suspension, a nice a paste, an jimmem, a snppository, a j'eiatinis comosn Low an ai0 I a l, ornopo r comprsing the sNAG nanoilbers is f'ormuLated as an ultra-thin membrnme, in some embodimets q composition comritmg the s\NM3 naniotl ., i' fwndtaed as a streSams a mat or a. bandae, solid fbrnaulationts suitable for solutiorte nor \uwpcnslo 2, liquids pror to adrnmisration are also comteoplated, is also possble that such conpoohions are mecorporated in or cated on unplamnabie devices. such as orhbpe'die hplams n ru np knee, shouilr, plus screws, etc , cardiovaseuar impanis (tem. cathereis, etc. and the he whete the amibsetria acti vnyi wn 'Ould b of'benefit, [00112| A composion comprising the sN\AG nanoilhers may include one or more of phiarmacem ially epumbl ex ccipient.it able exc~piemts rmc n tde w ater, saine, salt solution, deurt'ose, iyeerol ethanol and the like. or combinations thereof Suithabe cxcipients also include starch, glucose, l:actose, sucrose, gelatin, in , rice, 11ow', chalk, silica get. sodium stearate, geronimoiostearate, oil vien chi ding qh ot ptroleum, ammL 1egetaebl or synthetic orient uc. a eant oil soybean oit minyraoil sesamne oi0 and the like), talc, sodium chloride dried skimt nauiL propylene. kyeol and the lik. In addition, a enmpositn compnrising' the siNAG namotlbers may include one or more ofuvetting agents emulsfy ing agenta, pHT buffenm;ri agents and other agems. Inc sNAG nanotiber composi tions may also be ioeorporated ism. mpykn O34 m a phyOgica I v acceptable carrner Or example in a physiolog calle cmarrie suitable tor topical tpphcatton The term "phumaceutiealiy aceecae'' manS approved by a euuaor aenev of the l'ederal or a state povoCrmnt or MUted m thn U.S rmacojra or other isenea aliv rcce~nuh'ed phldrnracopeia fbr use in ana>A aid ooe.e part'dlv in huadns. Examples ofsoable pharmacemi cal careers are described n "ReImmton Phairmaceutcal science by 'X Martin. 1001 13 T he final VmouAt f the &\AG nmaoibers m a compfShnt umi uary, [Or F xampl. the amount or the sN AG nanotibers m a comnposimi (ev gn repared tbr admminitratumn to a Ant i (iny be areai' than or e o uai. t abotP'.. about S0% aboly '700, la 75"n. bonut 80% about 85%. about 90% about 95t about 2' , ot about t 9lt n "Ohi m vo 1 re, in one embodiment. T amount of the SNAG adofber ii .Omp0osi is about 90" "hoit 98% about 99. or about 100% Aso, the amount of We T\G nanofibe r in a compontion Mey prepared for administration to a pattent)n may(v aboutl MI-04,t about 60 .91009 about 70"'& 100% abot 7594100". about l"-~100%, about P0tU-100 sbout 5*0 00% about 70%-~ 95% about 75i95% ablout 808&95i <about 9OAA abou 70"2%-.( abot '5%'-.0% or about 80-~90%4 int*ght/voahnte A OMpOKion may comprise TOre tham . S0' t 60%, 70% 75%. 80% (10%v 95% or 99% sohan of the a\AG nanofibers 00114 1 A VAG nanofiber conpostion may be formulated into a wound eeing In ctain embodhnuem~s sA\G n anofiher composition i formulated as a ound dres ca in th oton of a barrie C memrane, or a fim. \oternativelv a sA nanofiber compotion may b' added to drong back dngs1 such as barriers. mnbrane, or Mlus. A barnor niembrante or n Ilm e supplied in a variety of standard sizes. wich can be further cut and dtd to the aTea beina trcaled The baking can be a csvmemional drevsn'u masenat such &s a bandage or gauze to winch a polymel or fiber is added or ooated on to to aphca io fth rtleNn atnaivc th A AG nanofibers can be formulated as a taier membnm, or im made out of tron rieioh ads, interospberes oromneroibads or toc cormposuon can be fobrmuat< d s a barmer formin : mat in certain embodiments, at least 75% a lat least 90% or at least 95" of a drewing is omrosed of the SN AG nanotibers. in ccna.in ap a drosw noes not contain a convMMtona drWing material suCh as a gau/c or banda#e Tnsuch emibodlmeN the s\AG nanotibecr itself is t'ormulated asna urund dressmgIt 35 100115 \ eOmpQ,,unm compns the NAG namoiber, may tarther o upiie any stitabl' natural or syMhenc polymers or fibersm Bxamples o0 Uitable polymrs or fibers include cellulose poly mers anta i po a i Wides. son! W; lik, v pol;varinizdes k-no vanunc es, pvnmae p des polv hdrades pwil lrodaole eosovbnzexoles polvester/anudc, poliester mdeirnirbleaide ''M poerbntemne plySultone >tamidQS. poypult'one nmdes and the like, copoly mers and blends thereof Other suitable classes of polymers or fibera include polvvtnvledenc tiuorides and pohyacry lonanrles Examples of these polymers or tibers inchude those described in US. Patent os RE 3035 U1254.0, 473 3 41 . 411 7 . 4 Y900; 9 : 43 1505: 4'0,41 ! 4 6 M9 4061, it and 'uropean Paent Apphction 0 2 i S7W all of which are incorporate d b iecerence, tih polymer or ftheirs cn inude at lest one of either of cellulose pol mers pobyrmids polaram ides, lyamid' mide's O> polinudes. in eeintati emodltenta dhe vollywie or 0ibers include polvaramids polester. urethan and polAetra fluoroethylene hino embodimem. the comlposi~n desertld herum comprise more than one 1pe of poy mer (g, tie sNAG nanon rber and cellulose ) (001161 In certain aspects. the sNAG naotifber is the onl acti e ingredient in a composition 14011 71 in other embodiments a coipositon comprises ne or more ationai aem e ingredients , to promote an antitbactnal eft'et and or healmr ig ound hobnw, In some mbodfimets, the addmonal acve ingredient i one or more anpW actenal agents eg, an antibiotc a delimsim peptide, a. defevnm-hike ptide, or a Toli..reptor Ihke pentitde t, or a growth factor. In spectic embodiMent, the addiAtinal acte ingredient a grow to actor such as one or more of PDXGF~\A PD)GL -\, PD(GiP43t PDGF~ ( 1 PDGF-DD. RGIF V2G .~ F6I. F>Gf -, FOE-'0, [E. TG C.$EG ?4 amphireguli eir'gulin betyeeKlulin, neuregbuns epigen, VI GA, \T4GM. VEGF-C, \ F- \F -' p 4eta grown t factor (PLA AE) angiopoleun- , angiopoiet v-F2. -, fil I L h1. tvocvte grow th actor ( H ). and miacrophagenstinulating proteicN ISP) In other embodi s the additional acti e ingredienm is an agent that boost the immune system, a pain relief agent, or a feyer relief aQgent. 100181 in certain embodiments the additional active inAredient is3a antbione of one ofthe fhillowing classes of amtihoties~ anerobides (cs " -vitmmyvcin, azihrornycn), atnn2gesides (e, amikacin, gentanici> neonyini, srepton kc t Cephalosporine (eedroaii cefaelor, eitamnve, cef ttpime). fuorotuinolones (e~g., ciprofl eim, leofloam a nenieiIhns (eg3, pemedL.i hnii amapicil m, amnourciln k terae elmes (c z g tetracycle:. &rsvyyl we), amd crbam ~in'fl e meropenenv inipenemol In some C"eife moodtmemt. the additonal active mgredient is one or more of vancomvmein, sufa drug (eg., co-mmoxazolertmethoprm sulinmetboxazole . etracvch nce. eg;doceehobnemmocene),e ldamrvcm, oxazoldinones ie g , linezolid) Kdapitrmycin, teeo planin, pmirlstn/dai t'oristmn synercid), tigeeveimet, aihcin, baci traciu. mitrofuiantoini bydmogen pcroidc, nomobioenm, netilrmtc, mem' uwlol. bee defensin-4, tobram\ycin, ehlor'hexidine diuconite, chlorhexidmie olueonate, lelvotloxcin, zinc, and silver i some embodaments, a composiuon compare, the 3AG narofibets and an additnal aemiaC ig e nefe c to treat or prenem or conioly used to trea: or prevent ao S. aUre howfceen MR S A infection, a Pi ~mw nriection, or a C dh de infection (eg an antibioti eflectivt mams mr O commonly led ajatt such tn'ectionas,) 1 m01191 A sAG nanoiber composnon may contam eolagen sN AG nanofier tomnposonon does not contain coansen. [001201 In econam embodiments a A nanohber 'omiowon does not comprsc any additional therapy c In euan embodiments a sN' nanobei composition does not comprise any additional anti cterial ten ent, a det'esin popi'de, S defeniin-hko peptide, TOiuKempter lke peptide, or a growth factor in some enibodenlnN j NAG nanotibe composition does nt Compose an antibotic. in. vet olier embody mtns a sNAG nanofiber compositton may compe an addional tmerupV gex. anl anbioticy) 1 one aechcimbodunemthe additional therapy tog, an anbiotiC) 1i not enca>ale imnobihnd or fbrmulated in tle sNAG nanofIbers ( I in. other aspects a1\NA nanofiber compositn does not comprie a siAnicant amount of protvem mnatcn in spcii ".mbedunents thte in.m content or a sN itaunobti compost i s no r h rs In other embodiments the protein content of'the C 'mpOSitiU.is vuldetrtabie 5\ (coomassie stanimg. 1001221 In one embodimem, zinc i also dncued in a sNAG nanober composition. i addition to its antimicrobial properes zinc aso plays a role in nound heading (see Andrce er a , 1999. Adv Wbund (ie 11 137 The oein is preferably added in the form of a sait, such as zinc oxide, s .Ae actate or znc gluconate, 31 5.4 Aaolakil ie N4 ,~ xG C nnw fitns |00123j A wide venetr of beetenal mtectin and doeases associated therenith inay be treated and/or prevented by the AdmiistraitH o the 4 AG native COinpOstliflS deseTed herem (sec. e Sections 51 and in 4 ir 1I one embodiment the ompositims detenied her em are baeteriosian.te n another anbonint, th opposition described herein are haeteneb~d t in dln enbodmcnl the cowlThions deserthed lereIn 1ay Ue Used to tredt ad/ 0 prevent infecions by (nam-postie bacteria andor a diseases associated therewith, I aun't emxodiment the comnoostiuis described herein may be u dS, to tecl1ti n by irainegatlie bacteria and/or any diseases asociated there itW in vet another emabodimnem, the eomposustois deserthbd heremn may be used to treat and/or pr eent mriectorf. by both Gram-negative bacteria and Gram-posiive bacteria and/or any diseases associated therewith. k001241 Baeteria infections that may be treated snd/or prevented using compositions described herei.n include inft htas by acteA of the ymrrilow trml, imntrH/uwn famiky ,sorcsbacteraccae tfumily, ik 1cridacea t'ait xx artnell Npetes. ;delbna l nmily. Campyhdaczscr seer's (hiaurydie Spe g i m'n ,C md uan kowoi ndi in intrb e~ ca fanvlyg. b OY)Gnh:z r kpecres. l'dn ard ella /'nh ocra regene$, trw/nma species Estcihcrcel/a ccl/. Hufnia species, Klebrieila species Morganela species Proes vadgarl, Provideint, .Siimondiv scsg SIrw maC e n's and Sb/:e/ia !ecr: L G/ardleIa familv. Haemnopiln /nihsenireo Htrdobaeerraccae tiatmHk7. Heloaer f~ amity. Legkona/arte ramily Li Ateri species, A cthviccccn ee family, mycobact terA teo , ca y ikw) aos p/ri/n heaild y Parson re/hlwaaa Pim umac& foccuv species r coonkvwas Avi.h R hiz b/a eae famirity, Spi//tan hmoil y, Sp~c r'olfmweaU Lamil v Staphyhlsae ms (eg. rmethied Iin~ restant Sitaphyiccvcon taceua arnd Stpriumctti pl rPug&st I S)tret~co< cm t$. Sti rvo< acu <nert, l Seta em ja 4uz a1S and Ytetto<u pneaiomcti. LI'utmpiri S/I' 0/Pi hater fatad/Or lamp/rn trio fanily. In a speci fie embodiment. diseaszn caused by or associated w ith infecon\ by such baeteria may also be prevented and/Pr treated usin thel comlpoutionS described herein. [00)1251 Bauerinal infections that may be treated and or prev ented using comnpoitions described herei also include itetions bx bacteria of the ollowing genuse: dord ella, Ronrcl/a. Erucella ci(am Cw npioixair Clamn d and Clamdnap e/ia. Clcridinm8 Craeu ntrcccu, Escheich, ncisela . aophius, eIbater Legiomdia4ht, Leptoospiea. List Mcubacterum, Mycopa, N eera Pas, R icetts ta. Salmonella, Shiigela& Stpylcccs Ste'ococcus Treonma VIhria, and /or >ersinia, ha a specific embodiment, diseascaused~x by or associated with inttetions by such bacteria rnay also be prevented andt or treated usm th c aomposittons described herein, 001261 Bacterial infectons that may bo treated and/or prevented using compostons described herOin Minde feton by baeria of the folowing species: Bacillus antArMa Bo a p s oIQa buror rn Brucn absrtus, Bruceya rams. Bucella imetensis Brucella suc Cmp Aacter jju. Chamyd bpneu mol5WnW!a, Chlamydaw dr cnmwarin Camidophila psittac. Clostridiu botunum. Clot riium dhicule. Clsztridium per trinns. CMOstridhum tetan. C jorebacr i um o dihthriae Etueoccus /faeaiis,? Eneooccu /eccium Ec:henric:hia coli, Francisella tularensk heninrphilus infuenae. lhicobacter plork, Legionea pneCumphiCL, Leptspira pn~eumpi . LeJpospira inlterrogankfs. ListerIi ocyl tenes IMkucoateriu m tae, M/coc~steriumn tuberusis Myfcopiasmai ponumonie, NVrsseria gon~sorrhoeae, NVeusseria mningilldes < Jserudnunas aerugi nosa, ProItius mibliWCfs, RIcktsla rIci0es2 SIona typh4, SOella p himteum, Shga o nnei, StuphyocoCculs aureu$ Nrtaphiocos e epidernniU ' Manphpi'ncus .roptyhvtti' % S yptocoet agdaune;O Stept.- p umona Srp t ococ 5 p Vo ep m p du oh A aa aond/o /ors est i a syecific embodiment. diseases caued by or assa oated a:vich. infections by such bacteria may also be prevented and/or treated using the ornposhintsn described heoin. j1)127j In some embodiments the eomositions descnbed herem may be used to treat and/or prevent ierions by aerote bacteria a nd/or ai enses assoated therewfih. hn other emibodiimems the compostins described herein inay be used to treat and/or pree n m ections by aarbic W bAe W on brd a and o isjoascad hrnt 2 h certam em'lbodimenlltS the compositions dencribtxd herein ane used to treat and or prevent P% ~e/ftdOmoS dterOIIW)a ofections .Pscud~om i.s a gramnegativo. aerobic bacteria found in i n ata 1r, otber moisi em" iroupments, plants s im hals, cliiedt .wohates of which produce the bluejgeen pigment pyncyain and a characteri stic sweet ordor. .Peudomoanas acrugintmM is known to cause urinar\ tract mnt'etions, pnuurnma respiratory system inutbtions, dermatzts soft tissue infections bacteria , bone and joint infectionts castrnestcaz iuftettons 39 antd aa vanew ot svstenmc ifetios.t li know to 0e an mportarrt cduse of nfeti on parucularh in patonts wt bumr, pans uih qcstic fibrosis patients who are irmimmnosuppressed (ey AIDS and cancer patients And in patient who have been hospilied tr 10nger than 1 eeck. t is a firequent cause of nosocontial illctioris such as Ot not limited to pnieumona, arvd\ ir act meclions and bactermia. An one. o all b these in fections na be prevented and or treated by the corposntlons deeribed herein, 1041291 in oie embodiments the comportions desrhed herein are used to treat ad/or pre\vent V a' maetnao, and purlcularlv, SJltTh hTu da weNs ionectios Use of a WNAi iunioiber comlpo.4tton in this emubodheodoermounentl~ tX~ ) im describedt herein mnay precude: the generatim of resistant oignisms as well as allow for the antibo e-ndependent clearance of a bacterial infection |041301 hni eritain embhodhments the conpositions descnbcd herein may he ased to combat bacteria that are resistant to one or more aubacteal agents. For example, the compositions described berem may n he used to treat bactei a tba dr distant to one or more atitotes fo evMNle resMtant to Conventional antibotics su aNIs W RSA (methieillin esistant' trto <crum Penicilln-reistant Lnsrecyya.> i - i S Penicilin-resitant yrepo ca pncu.9nh), isoniagid/ritampin-resistnt MikcobarterWthweoufr/s and other antibiotic resistant strains of hacteri ite , re~stant strains of /:-vii hhnonnd hr (XnprXi/wle r, and sreproces i in one embodiment. the compositions diaclosed herein may be used to treat mnaltipie drtg ressunt baeterria 1001311 a n ome specflce eT bodimentA the com!ositionis described herein may be used to treara Ador prevent MeilcedlinnresNiant Stapnyrh nccs aiL' '\RS A'; it may also he called mntidrugresistant Sapow-item war or oyaciiin-r esitsan &aphvbcctr creur ('ORS \N. MRSA is av strain of Sapvkaos aqvreu that has developed resistance to beta9 latain antibiotcs, wVich inchde but are not hired to the pemedins (ponciln, methicilim dieloxeilbn, naeihin, oxacillin, ete.} and the erhasosporins Some of the known strains of MR.SA inlde 15.RSA $1 and EMRSA 6 (also known as MRSA2 L 'hi are riston to eryhromyein and iproloain; (TV (also know n as ST&USA3O ST I S \WO&400. STIKUSAO.; STJ l'SA OO 1ITV strains TsO strans, and ST9 srain, MRS/A is responsible for a number of infections in humans \lRSA is a serious heLath concern, causing 40 approximately of healt-care associted staph fetions. in the Us., more than 4000 people de elop serious MRS\ infenon and about W X000 die romn infection each year. Fspecially prevalent I \RS, is in hospitals where rnsk n'ceors tr MRSA inton include pr antibiouc exposure (e g. gumnolone antihmotes), admisson to an intensive care ut, sumgery and exposure to an NIRSAeoloni'ed patent. Patients wit open wounds imniunocompromised ptn(et * H VAD caneer tranplant procedure severe asthma) yung cidren (ve human mnant and human toddler, and the elderly (e g, elderh human) are at high risk of derwelopmgian \NRSA iNeton. iher rsk rates for NRS A intfeton are also observed in nen~on drug users persons wuh diabes, tients with derntologic condittns, patiens with spend time i confined spaces (prison inmates, soldiers. pat ems in long-term hneatr cihitir, sch. asa umy homess, £ 1 niusrost commny eolnit/ts the anti mares (the nostrilst altru test of the respiratory tract, opened wounds imravenous catheters and nrrfat iract are w so potiial sites for inifton, \'ost ofeonfunit)-a tdl NIIR.SA intfeetions are tocaed to the skinD ald sofl tissue The intialsvnmtom asot MRSA\ include red bumps h hat rsmble pimples. spider bies or bols that may be accompanied by fever and rashes; the bumps na later dvelop it& O Pus-filled bods (omson manitestation Of commty associated NMR SA are sin nfectios such as necroti;ing OsWNo\ or pyo ' m ti- werowiiing pnenuionia, ifeie endocardis, bone or Joint nectiok Some MRS \ads ne and toxic shock svndtom, whih may be due to tois arried by such strain (ey. P\ I PSNM MRSA may cause elluliti Any oihe abomested or known i Ite art stream\ of \1 R\, patients diagnosed with l RS \, uptonms of NIRSA patient populations at risk of \MRSA\ and-or diseases associated wah MRPS A may be rned with the compositons denbed herein in some embodiments the compositions desenbed helem prevem onset or dcvcient ofone or more ot the symptoms of MRx'NA o" reduce turation and/or seerity of one or more of these sy mptoms (eg, synptots described herein). 14003 'he compositions described herein may be used as bactericidal agents to ki or daage unwanted bacteria For example, the compositions described beret may be usd to treat estbihed bacterial infecions, prophylacually for the prevention of bactenal infections, or admistered topical to areas of a subject that are susceptible to infection or to area of the 41 Nd at a likely hw FlY baf riagrowth ( ,nfeh pthep ounds,, bed sores and I (0d1331 in certan embodimnts, thecopoiosdsrbdeeird ebatiltzwt andor buateril survial by more than about 0 1 log, 02 log 025 0g, m. 0 C4 log. 0 5 g, 0.6 log. 0.7 4025 O ( 0 logp 02 i og, .25 log, 1 5 log. 1.75 log, 2 log, 2.25 log. 2.5 log 225 'Lo, 3 log, 35 lm t5 log. 175 og 4 lou 4,5 log 5 log, 5.5 iouo o ny a5 hou 7 log, /.5 log, 8 log, 8.5 log. 9 log. 9 5 log, 10 log, 10.5 log, Ii 1G, E log. 12 log, 1> lou 13 log. 13 5 h 4 log, 1 4 5 log, or 15 log of colony tirmig units ('Im in reruam embodimenta thea comnposWions descredbeein reduce bacterial growth and/or barcerial SUM Al by about 0.2 log to 15 log1 0.2 log to 10 log. 0.2 lo to 5 log 05 log to I iout 0.5 log to 5 log, 0.5 log to 3 log, 1 log to 15 jog. i log to 12 log. 1 log to 10 log i log to 7 log. I log to 5 log, I lo g, 5g log 1s 2 log o 15 log. 2 log to 10 log, 2 log to 5 lg, 3 log to 15 log, A log to 10 log gn to 5 log, log to 10 log log to 8 ig, 3 log to 8 0 og t o g log to 7 log, 3 log to 7 log. 2 log to dlou f cokylan ng unat (CFU}ml andany value m bin H n these vale In CO ram mbod ic ment th e compo'tons described her reduce bactal Ioth bndor hutermai \urvwaI by equal to or r- h a 1 1 10 2 > 10, 2,5 x 1 D x 10" 4 x 10, x 10, 7 x 10' x 10' 1.5 x 10 2 -10". 1 x 10" 'x 10, 7 10' 8 x 10 1 x 10" 1.5 K 10'' or I x 10' (4 ') mL or Iany ra of values if betw een these n alusn in Norme enbodnmcm ouch r Coatan m baeitenial gruwmth n r ido i -v al is achieved in less than about 30 mnmuies I hour, . hou 3 hom. 4 hours, howr ( hours 7 hours 8 hou Q hours. 10 hours horn 12 hUr, 15 boon, 18 hour 20 bours .22 hours. 24 hours 36 hours. 48 hours. 0 hours 72 hour dy oe da wo days, three days tour days. five days sewn day, ten days. one week, two week three w eek - "our w eeks I mmb or 2 monhms after treatmem of a bacteal mfetion with a sme apphcation Jose or muhltie applicattonidoses of a siNAG nanoliber comnposidon) j101341 |IN one embodment the inteion to be treated ud a <'AG nar ofiber composition is not a vtal infection, a mingal infection, a parasite untecion or an xest infetion.. I0Q1351 A variety uf diseases or dieae condsons asoo ted a ith baterild infetions mybe treated and/or prevented with the sN\ nanofiber compomuono desemzhcd herein (see. e.g Section 5.4 . wf) In one emboen, methods for treating an estog bacterial eton or a: disease assocated h\ nlh a baer ial nfreton are contemplated, 100136 In some embodiments, the compositions described herein may be used to treat wounds (see Section 5A4 1, iri-at in specific embodiments, the compositions described herein are used to treat bacteriiv infected wounds. in other embodmnents. the compositions described herein are used to prevent bSctIA infkection of wounds, in some embodiments, the compost ons described herein are used to treat bacteria known to be associated with wound infections, and specitically used to treat Siahlcccsnru/MSSreMccu KJ~ms son!&UL toeuMRA 5tpt".~ pyrogenes, Entrcocci> and/or Psm ona a aeturusginca infections, and diseases associated with I0137 In other cmbodimems the comp rntms ieseribed herein arc noF used to treat w ounds In one embodimiem, the compia described herein are not used to treat chronic wounds In another embodiment the compositions described herein are not used to twat burn notinds i y et another embodiment, the vomnpositions descrnhed heremn ate not used to treat surgice woinds In one embody iment, the compositions described herein are not used to treat chiomte wounds burn wuids and su'eal wounds. in another cndboment, the compositins described herem are not used to treat a wound and or a bum in vet another embodiment, the compostons deserved herein are not used to teat m-ntected wounds> [99 1381 in some enbodiments the compositions descend heren are not used to treat : bacterialV infected nound. In another embodiment the compostions described herem ae not used to coal a b actena ofeetou associated wth or caused by a wound. In sone enmbodiment the: composnons described herein are not used to treat bacerina known to be associated w ith wound in c'ions such as Sintprocoeu neiM \'1RSA. Srpwcj4QCQCai flyp , . emnanOC andio itkPakons aurlPgmowsa 100139| In some embodiment, the coinosiions described heein may be used to treat or prevent a riety of bactenai ofections and diseases camed by or associated with bacterial inflections which are not associaed wth a wound see Sction 5A Z ;r ha, I1.401 1n. certain embodiments trcatnteut of a disease assocIated with a bacterai infection compares administration of one of the compositions desibed herein to a subject or a population orubjets to treat the disease or to obtain a benefOiCit otherapeutic cieet in specific ermbodiment such treatment aevc\es one, ttwo, three. our, five or more of the folowino effects in a subject or a populatin of subject i) rduction or ameoration ofthe severity ora disease or a smptom associated thrcrwh. (ii reduction of the duration of a dioa or a symptom 43 associated therewth; (in) pre venion otthe .rogression 0: a Josease 01' a SYmpitm aSSOented~ thereWi h i i regnreion of a disease or a Symptom assoeined therenwith( i prevention of the deeopent or Onet of W the recurrence of a ymptom asecated therewith: aii) prev mention or reductioni o' thf sread o a disease t'rrn the betor population of subjects to mioihe suict or popuation OSubi6XtN (Viii) reduction n organ filu asocated wt a disease' reduction of'the meidence ot'boaplt autm:( rcduction oF he hospiabiation lenl 0) an increase the suiwivat: ii) ditmintatiotn tot a disease: inm) en hancemnent or miproventent o1 the prophv tactic or ther apenue effeelt s) of another terapy; (m) improvement in iahwy of lie as assessed by nthods wnel known inte art, . u questionnafrc; (\v) reduction of the nnber of symptoms of a disease: anor (xi) reduction in mortality in some embodiments, treatment comprises any thapy usmg eornpoaitions deserihed hereim, |0q1411 in ome embodiments treatUent or a bteril inction comprises adomistation of one of the ompoitnora described hockin o a ct populu iofoi cubjectto treat the bacterial mNfetion or a synmtomn of 'atatkd nfection In specfie enodiments. such treatmet achievs oiet, three, rour, nic or m or or tn ikaw i effeecs 'n a subject or a population of subject (ii the clearance of a bacterial infemetin, T Ain radltcaion of one or more symaptonms assoctated \ith a barttr it 0fectionjii i) thb rduction of inne reguired to clear a hacterial mneiorr (nv I the reduction or amelioration of the sesenty of a bacterial ifction and# or one or motre sy mptom>s associated therewiath: (\vI thC reuton in the duration ot a oacerial infection and/or' one or mo~re svrmtnspu assotatnld there1tth a( {vi the prevention or delay of the eneration of a t rsttant strain or straims of baeteria or reduction of a number of reuistrains of bacteria eratd: 'a ) the> preventin in die reenrrence ot one or more symptoi as sociated therew\ith,:t viii ie aedUcbon or ebanruaUon in. hehaeterial eel populahodi (Such as reduction in bacterial counts es n a biological sntle ofda patient, as measured by CFU/'rmh or a log r eduction bw one of the nhiods know n mo th at or deacnbed bereni, ; i) the reduction in hospitalization of a submit' u the reduction ii hoopitalization jenah, eec the increase in the survivld of a subjer t: i the enhancement or rrprovement ol tbo m er~.ap.ti efflet of another therapy;w (iiu) a reduction im mortality (xt ) th. reduction or 'hmmitton itn the spread of the bacterna frorr one subject to antothet' Subject or one organ or tiss te to another organ or tissue: (xv) the Preventon of an increase in the tnumber of bactera, (xi ithe prevention of the NOW 4C deoopmenm or onset of one r more symptoms awseiated thew (n: m the reduction in the number of Symptoms ssocied nwith a bacteun infeo t on iiv the inhibition or reductn in producton of hbctenal lox in or toxins associated with a bacteral iftciion: (ix the. stabilaion t rcductin of inflammathn associate w ith a bacteral ifcton; the r edcton. in organ uadare associatd wit a bacteal infection or a disease asceice theren ih, and or (\\i) inprol innt in quahty of lifea.s asced by methods well known n the art, j&; a [ 4'%j in certain embodiments, admmstraton of the copostiums desenbed here to a ub t resuulIs in one or iore of the following II iO e htio of the expresiOn of one or mor defensin proteins and/or defensinlike protein, n i) the induction of the expression of one o more Ioll reepor and/or il the induction of the expression of one or more proteins that are baticial for cleainem or reduction of a hatenai n lion or one or t symptom asociated herewith f001431 in ceain. embodnmems pres on of a baeteI i nteuon compises admmisraton of one of the composions dwscrhibd he t a snie a u a populatio of subject to acher e one or more of the tollowinig, 0fter () the inIbitio of the de elopmen onst of acterial infection, or a syrnptom allocated theren ith and or ii the 'himon of the reurrence cfa bacterial mfection, or a symptom sociated there ith, [001441 In omer embodiment prevenion of a bactenal infection comprises admmnistra on f one of the conpositions descried hrem to a subjct or a population ofsubjeet to prevent a disease am oriated with a bactrial infection. In specific emRbodthents such prevnen achieves one or more of the follow ing effects in a subiect or a population of subjectrs (i the inhibition of the development or onset of a disase asocatd wih a bacternai metion or a symptom thereof; and or (i) th ininhio on of the recurrence of a disease associated with a bacteral infecn or a synmptorn associated thlerew it $A1 Treattnnt or Prewvntion of Raeteria infecti in Xonnds 10145 in certain emodiments, the s\AG nanotiber b Onpositions described herein may be usefultor treating a w.e ariet of bacterially infected wounds a any tsue ot the body or prevntung infection of wounds at rsk of becorin infected with bacteria 1001461 There are two tpes of pounds, open and cloWs open wounds arc clasified according to the objec that caused the wound For exantce incsin or meind wounds 45 (i cdg surgrcal wounds) arc caused b\ a eltu sharrp-edyrd obseet such as a knifi a aor or a class splinter. Lacerations are irregular woidOfs caused biv a blunt impact to soft tissue which hes ovr hard ise t maceration of the skin coVerA the skAll or tearing of skn and other tissues such as caused by elidbirth Abraiols or graes arc seril wounds!nn w h e topnwost layer of the skin (he epidermis) ! scraped off. Puncture ounds are caused by an ol ect punctrin the \klni snuch as a nail or needle, Penciraton pounds are caused Vn an objet such as a knife eminn the body. Gunshot w ounds are caused by a bullet or similar projectile d rnmto (a enur wound) andlor ihmugh the body {eg, exi wounds in a medical contest, all stab wond and Gtnot wounds are considered open youtds, Open wounds also inclde burn w ournds induced by therml chemiCal, or electrcnal injury. Closed pounds ielude ontusions (more commonly known as a bruise, asked by blnt force tauma that damages ussue under the skin, hlniaioma so called a lood tumor, aused t iae to a nod vessel that turn causes blood to collect under the skin), and crushing njwi,'eaused by a great or estrem arntont of 'orce applied Over a long flenod ot lime). [00147 1I certanM embodiments, hc conpottion% descied herei are used to treat a bacterial in eed open wound o prevern a trA na doctn in an open wound, in certain embodiments the compositions described herem mu be sed to treat or prevent a bacterial infection of a gunshot wound, a puncture wound and or a pnetration wonmd, in certain eimbodmnem\tue comlpotatros desenbed herein inay be used to treat fir prevs em a plos~oyeratave bacterial inetion, a surgical site bactenal infction, a o-tter-relatedatciai if uan or a benyialybi s-retied bacterial intection. in yt~ ihr .. emboditnent, the 'olnipusilhons descrihed herei are not used to treat or prt ent a bacteria infetion in an open wound, a gunshot nound. a puncture wound uand/or a pt*11tiration wotind in t. ert em'bodimenits, the compositionis descbed beremn are not used to rreat or ereema pot OPeratnve bacienal ifection, a surgical sie bacterial infection, a catch etrrlated bactertalinfecuin or hemodialysi,-re'ied bacterial infection. 100148j In sonic embodiments, the wound is a chronic wound, Chre wound can be my wound that ,ad, to hel pReperly including a surgical wound (e g., a skin grafl donor sie a cutaneous ulert i e a diabetculctr a i eous stasis ulcer, a jeg ulcer, an arnral insufficiency ulcer, r a. pressure ulcer, r burn wOund in one embodimenit, thlecmpositions described heremn are uscd to treat or prevent chronic w found ictions (i ant infection associated with a 46 dhabetic ulcer, a wenous stasis uler, a leg ulces an arterial mnsut'iecv ulcer, a pressure ulcer surreal wound, or a burn jn vet another embodient, the comoitions dcseried heren are not used to nteat r prev em chronic mnd bacteral inc ons e g. not used to prevent a bacterial infeion associated with a diabeni ulcer a. venous stais ulce, a e ule, an aruterl insufficincy ulcer, a pressure ulcer, a suruea n ound, or a burn) 1001491 In certain embodimets the ompositins deribed eei mre used to treat or prey cni nosocnmid bucetri al infections, Ot' the nosocomid ab acterial in fections. surgi icat wound ho xwalhwingw"NOM mn bacteria! ofens predonmate; o stat ties h m up to $ .-tall surial patiems The direct cos Of these types of infectons is uppreimately 4 5 hilion dollars per year Many of hospitab-contracaed bacteria developed resistance against anotics. and thus non-antibot based treatments are deAired, Use otthe sNAG compositions deen r setting could defray m bch of the cost and rnurkedy reduce tih' prdduun of anubiotic resat species in yet another embodiment the composition dcsenxbed herein are not used to treat or prev ent nosoomial bactr i uouns, such as si ga ~at bac'teiasl intecuinas f0f1501 In one embodiment the corposition\ deserieed hdcim may b. used to treat or prevent bacterial .ntir ons iu bleeding wounds {e blecdng surface wounds) I one embodiment th~e compositions described herein may be used to treat a gun hot wound, a puncture wound, a penetraton wound or a suTgic T found in order to treat or prevent bactenal fecninn such wound, In ye another embodieWt h compostiOnS described herein are not used to treat or prevent bacteria ofections in bleedig wounds (e.g. bleeding surface wounds) 001511 Thc compositnons described he may be usful lir treating or preveinng a bacteral iofetion in cutaneous wound, such as wonds affecting the epidernal and dermal layers of the skinas wel as injures un the corne and epithehAned organist ioder to treat or prevent a bacteial inibto in such w ounds The wands may be caused by a wide vaneW of physical trauma, including cts abrasis, burns, chemical exposure. surgical procedures (e.. utgical m ns skin grading) i one emnbodient the compostions debad here may be used for treating cornea and selera. wounds including wounds which at'et the epithel a! Layer, stronmal layer and endotheial layers of the eye in order to treat wor prevent a bacte;rial infein in such wounds, in yet another embodiment, the eonpositions descdhed herei are not used to trea. or prev ent hacteri alinec tions in ltientom wods 1001521 In some embodimens, the compositlons described herein may be sed to treat a wound in a patient diagnosed with a bacterial utlinon. In ecrtio emnbodiments, where the conpostions descrbed herein are used to treat a bacterial infected wound. a wound is determined to be bacT inised by a tfst or an asa fr the presence of a bacterial antigen, in one embodiment, a wound culture is performed to detect a bacterial infection in the wound of a patient. in yet other ernbodiments a wound is determined to be infected due to the presence of one or more SYmptoms of bacteri infection. 1001531 in other embodiments, the compositiOns nescribed herein mav be used to treat a Mound in a patient when a patient disphys one or more of the sotm of baceral section such as: a wound is slow to heat heat, redness and or sneihbng at the 'ite of the wound; tenderness at the Wne of the wound; dratge of fluid or pus at the Wt of the wound and/or fe er Symptosn of nound bacteral ofection mel&de but are not Inated to lTalied anhenma, localbred pain, locaUed heat. eeiluhtis ceem. abscess. discharge wMUn may be viscous. discoklored and puruet delay ed of w ound healing, discoloation of issues both w nm and /or at the wound marins, friable. hNecdmng taranulation Iabnormal smell c~tog from the wound site, expected pain andor tenderneo at the site of ge, lymphangtis 6 e a red ie oriwtnathg from the noud and lWadng to su oln vender lymph glands during the fected area and wound brekdown associated wt wounroM bapo orwoti w ound develops sig of gt ranulation uiu r' th base as opposed to a unmiorm spread of granulation ossu& uta\ the whole of the wound bed E in sonc emodnents the coniposinr described hereim prev ent the onset or dewelopmnent at one or more of the abow isate A smptorra, or reduce duration and or sevrity of oneo or more of thesoymptonm 10Q1541 In embodimem, the eonposinous ibe herem er hearing and treatmem of a wound bacterial mec o o for wound healing and prevnon of a wound bacteria l infection, in one embodiment the compositions described herei are used to enhance wound healing while concurrently treating or presenbuU a wound bacteral infection. lffects of the sVAG nanoiber composition ou wound hea wng and some of the uses of the sNAG nanofbers in wound healing applications have been dscrib':d in T5 Patent Pub No' 2009/0117175. which is mneorporatcd by reftrc h n iretyseect e g. Eam . 48 5A2 Treatment or Prwvntionf Other Rcteda1 infections 1091551 hi certain embodiments the sAG compositions described heremn may' he sed to treat and/or prevent bactena! infection of the skin, pcastroimsntac respiratory tract. urinary tract. reprodttie traet bood throat, ears eye, antus or an\ other Ot id; or Uise of the body. in another embodiment he AG naroiber compositions deserhed here muy be med to treat and/or pre ent sn conditions. gastroitestinal conditions, respirator' conditolis and/or conditions of any other organ ortissue associated wnith a bacteria section in some embodiments, the sAG& natoiber i moto erbe ee r phd topically on the th A = 0;corniti4os descrhbd herei areap skin voul, ear, ey anis or groin areas of a patient to trat or prevent a bacterial infetion in some embod . the omposiuons deserhed herein are used to trea and/or prevent a aetenria infection of an organ or Panoc of the body that is not a the s ite fa. wmdound Anor is not associated with or cased by a wound [001561 In certain embodiments, the aN AG nanotber compositions described herein are used to tra ans es iting bacterial infection. For example eh composition may be ubed to treat a subject dhapnosed with a baueeral ifecton by a test or an assay such as one of the tests deserhcribet ri or known in the art AIeMnadvely, sun hi nuy be used -o treat a subject d isp1lamg one or more symptom of a ha terii nfechon or a diseaw associated u a bacterial imec-tion, such as one or more symptoms ofa bactenal infection know n to a skilled arrtsa t' pg deevricnd by a treating dotirphyinn to bea symtton of a baetal infection) and/or de\Cdberein. |00157| in ai embodiments. the compositions described herei are sed tn roeat a co11dttion associated w ith an inmhala rie in h'scterial i'obiota or a con Viton itssoeiated w ith ar abnormal or alter"edx batn icrobiota, For example, such corusins may be used to treaty Ani eondiuon in a patient wbose skn bactenal mielobiota differs onm that m wiontnl subyj ehs i t , stubjects with no synoptoms of the skin condition) in other examines, stuc compositions may be used to treat an itestinal condition (or a condi ion of any other issue or orgapt in a tatiCnt whose ntestial bacterial microbiota 10 mi crobuota of the other isue or organ) differs from that in control subjes cit. subject with no s itoni ofthe sitesinal condittm) [0015Sj in other embodiments, the compoaayatehed herein may be used to treat any disease kjuw n to be associated wth or ascerbated by a bacteial inteen ieg. atine in one emnbodinent the . 'pt. derahed here main h -sed to great a cystic tibrosis patient infecred by P.t'Qse wSa, 1 in me embdiment% th cmpon Jre n effete nst oxins seceted or exeted by nactera. In one eniboditnent, the nmposions described heren ns be used to inhibit,/reduce a bacteri toin (ad/dor a cndiin or symptom caused by a bacterial toxrmu f or example toxins produced by Ra OdlaW rw~ar (iloestndnm <hc<de, t'ornetkrnafl. mm a sph r taupwnas rwtm a.cgndotoams, and/or the cytlits.n Some defen s siU are ab to tb th bacternal ton. in diong those produced by Ba/OS an:hracts (JoSWrW/d wanted Ckvybrrmcatem dwphthnoa Pcurmnwnaas aUnnaOW and the cytolyinB tendotorns produced y ranp ie bactera tht red biood ce A w ivn these rurcuons of defensins.activation of pathw ays resulting in defensinl expression and seCretiont ray alow for the Snhbhotie~tndependent clearance of'baerial infection, ard thus avoid the genellion or bacterial resistance. &1M6(| 1n certain embodiments. the compO5tion9 descrihed heremn may) be used to a. ut ador prevent , on or more bacterial inttbicn of te skin or dsctSeCof the skn assoeaedx uth a bacterial infection In Sa emlhodin>onvr. the cgpos' nons described hereti are used to trew, Incalized km infections and/hr difflse Tm inr'etions. h3 some mbodients, the composition dcrUbCd herein are used to treat or prevnt skin infections or diseases of the ski associated wilt) bactial ifctions aidteing the epiderm is. (r.. aid/or subetancoU s hypoderniA tissues ofthe skin, in somic of these embodimnents, the affected layers of the .i include One 01 nire dwcrx of the epider1uN (i ne, stratum basale, stratum spinsun. stratum grsanulosunu, strtuml hiidun, and stratum orneum X One or OorC types of tissues of th' ernts (ie., clladzen. elastic tiss, and retcular fibers} one or wore lakers of'the dernts (i , t pper, piipiiltr Naer aid the lower rencular layl iTdd on or more types Orti5sI of th h)podens (r S, fat, elasun and connective tissue> In an embodiment the cormnosition doweribed herein are used to treat or prevent bawevit! infeiu0> on the skin surtace t another norbodiart, the. compositions described herein are used to treat or preOnt j $chrivkacc Staphq') ifenlion of the skin and/or a Stp e ( ep'' infection of th ki i I t :mother embodiment, the Composition deserbd herein arc usad R) treat Stap/Qor' cm Wo/bs and or £p/wi<on ur io Ation ofthe skin in ome embodiment the compostiAns described hrno nre used to trat or prMICOceiluU imp xt, licuttis crytadsnd. edrhtucle, fuuncle. abseces er. sieandu/br a cutaneous amna i another embodiment, the eorn posrtlons desebe nerC11n are used to teat or pre\vent clliltis. C eilitis aff'etheo deeper derm)is and subcutaneous tisues, arnd usua li attects the face, arms and lees and ahnost alrways oc eurs due to a break in the skin that leads to a bacterial infecion. T'he symptoms of eilubtius include one or more or, s eling of the skin around the break in the skin, pain. tendernes otwnard sigtns et' bhstcrinrei ed inies between the ivmnh nodes fevr and lhd Is In another embodjment, the coniposition described herein are used to treat or prevent a. hetorial infection of the skm at the hasr foll es le a. ,b~ flicubtits The symiptoms of tol hctd include 5nWIlbUg :ptstules sturrotoding the hir, hard nodules and pan in <m other embtodiment. the comnposinons descrnbed herinm may be used to tr ea~t or preventt acne. A ppearance of acne is n equent lv a svrmptonm of a bacteril infection ln One em ibodinment, the compositins described herein are used to treat or prevent de'rinaldtis assoeialtd witlh or caused ha a hacelral iniettin In stome emnbodiments the compositions described herein prevent the onset or dev elopment oftone or more of the abo\ elistcd sanmptons, or redhuce duration and/or seve\t cit1 one or rnore of there symntoms (016I i certain emnbodhrnents the ormposimons described herein utty bie used to treat and/or prevent onec or more intesinai/dig esie baceraul in fertions or yastrointettmal diseases asctiated w ith bacterial infections T he commron fbrmas of intestinal bacterial infection include ra oneni a n F < anY. (Jcidhan Suptvlo.ur.u [is tcicr and h3>nic T hese bacterina cause diarrhea and infianmmation of the stomach and intesties atso knlowhn a aastroenetatau y~mnptorns of an ntestinal bacterial in ietion mnclude but are not imnted to albdtmil cramps and pain, bloody feces loss of appetite, nausea sometnme5 accompanied by vomiting fever, and diarrea i n.In sonme embhodtiments the sempomsit ions Jesceted herein arc used to treat a patieni displavini one or more s~tons of hiud wonwu w\heb i f tlen dshutcated with a backenal ink mton. 041n b i certain emod~ximents. the comYpnsitioi s described herein may be used to tredt a disease associated with a Staprninfection (e g , a Staph ex'vu ane rews infecton). in som~e ot' these embimnts, the dseas is a Vlagh ietion of the skin, nose, rmnuth, andwor genital area. In some embodiments the disease is p"nuona, meningiis endocarditis toxic shock nrome,> and or seprirenmia .1n an emnbodimienib,. the position described herein mUay he used to treat Staph bacteria resistant 'to one or more anitibiotics for e xample, nmethiei din resistant Staph 'Ii aucn ' ("M RSA' In certain enmbodients the composimons desired herein are admnster ed to a. subject diagnosed wh a Stan ofection, or to a submit displaving one or more symptoms Of a Sam? infection (0 e presee of one or more of smnaI red humps crsft red bumps pus filed bumps or abscess bols sites in the eves blsters andor red scabby A in such as red scabbx skin around the nose and mouth , or amptonva of a Toic Sock Syndrome. In some embhodiments the composhons described herein y ev nt the onset or dev elopmnent cot one or more of the above-listed symptoms or reduce duration and/or seventy of'one or more of these |010163| In certain enmodiments the compositions described herein may be used to treat or pres nt a cold associated w ith a bacterial section, Fo e>,Fnpie the compositions described hereim may he used to treat or prev ent a cold that persitst dv\sfle the use of standard pain relief medicatons. I one embodiment, the com onions ascribed dore are used to treat or preelt a bacterial nfection of sinus. ea or throat. T he synptomx of suc bacterial infections in cude Iocaed pam and ei g h aeter ; Iee no in the sins may lead to nasal diAcha gea nd acute pum in parts of the face or forehad in one embodmenu the compositions descrbed herei ae used to treat strep throat (at >n U nrw in sonc eibodiunts A compositions described herei prevent the onset or development or onor of the above listed symptonms or reduce duration ander sev ernido one or more of these symptoms 100164t 1n SOme erahodinemS the comnposions descried herem may he ued to treat or pre nt a gental, urinary tract orunad bacterial iWtion, or a disease of uriary or reproducmve tracti assonacd with a bacteria infetin in some or these embodintents. the comnosmons described herei may he used to treat a scuall transmitted disease associate with a bacterial n fertion> mptoms of such infreetons ms ude. ht are not lmited to painfWd urination, cloudy dai during intercout s 1 some of these emboimnts the desenbed heren are used to 'reat or prevent one or more of syphilis gonorrhea ciamy dia and trtchomnamdts. lI ole em bodin et,. the compositions diescrieet a'e used to treat Chiamidva, In another embodiment, the described herein are used to treat gonornhea poms of' gonorrha elude located pele pami , inte hn and irriation, pamful urination, a thick yellow or green diseharge, bleedwny between menstrual perods in one embodiment the compositions described heren are used to treat or prevent a. baterial gnos ineon Symptoms of bacterial ayinosis include ainal discharge, odor.vaginal itching, and abdonina.

oain r sO emboiment thecornposonsdescribed herein prevnt he onse or development of one or more of he abovelisted symn1m or reduce durationand/or severity of one or More o othesey thntouns 0164 i other embodtments the composioons described herein may be used to treat or prev ent a respiratory tract infecuian i e g a bacerinal ietion of the lumgsy or a respiratory disease associted wa a aterAd infection in some embodiments, such compositons are used to treat or prevent upper respirator tract infections in one odimnt, uch compostons arc used to treat or prevent tuberctlosA Tberculosis is caused by mucobactenur tuberrulos and it is :: hihly oetnous disease that is spread from peron to person by sneehg or saa, Tus, the compositions described herein may be used to treat not on, ijeets diagnosed with tuberculosis or displaying symptoms of tubercuoss but aso mdividals in contact with such sulhjects (e~g frtv members caretakers or mredical peraonnn B tv\nmtoms of tuberculoMis include coughing bood, excesive weight loss, fatgue, Vs of npete and persistent ver In somne ernbodineis. the eompostilota dsdribed herein prevent the onset or doveliOpmeit Of One or more of the aboveisted symptoms or reduce duaion and/or severity of one or more of these symptoms in one cibodwinnt, the compostons d ribd herein are used to treat pneunonu and/or a &ro ptc sn~ p}&Jtkilth menfectionl in :another cmbodment, such con positiotls are used to treat bronchitisn n one embodim nt, the composmons descibed herein are used to treat Aforr; <turjein KSin pb< <44f D, pneumo!C and/or tib mophdis in uluenza 101661 In somw embodiments the compositions described herein, are ued to treat or prev ent bacterial in echos of a imrosal suriake o g oral mucosa. or a diseaseisoniuom or mucosal surface associated wth a bactenal infecon. in one embodiment. the compositions described herein are used to treat or prevent bacteria: infections of the oral a I For emple, such c00010p0s00o5 marl\ be used to treat or prvent condition; aseciaed t ltiervdl inteedons in the mouth such as gingiis cares, ad/or tooth decay. n one embodiment, the copusions desen bed herein may he used in on3 bygiene roduc~s [001671 In one embodiment The compositions crpbed herei are used to treat a bacterial infriom of the ear, such as middle ar inetion, or d dsease associated witb such infection. n one embodiment, a composition desWribed here is used to treat otis media caused by a bactealainkection.

f09168| 11 some enmboduments, the ColpO~sitions described herein are usicd tO treat or prevent bacterial infections of implanted prosthesis such as hearts valves and catheters, in some embodiments, the compositions described herein are used to treat animal btes.r xapl cat or dog bite, in order to prevent a bacteria infection. 001691 The compositons desned herein may e used to treat or prevent a yarety o' diseases aociated with a bactenal mntection mcudig but not hmited to leprosy Hansen's disease) cboler, anthrax (ea cuaneous aritthax, pulmomar\ nth ra , gasimintestiAl anthratQ, pertustys glanilorna Inuharale, bacterial s aaU1051, go0norrhea, opthlintua ~onatoriumn, septhc arthnirs,~ syphh congenial Syp phii wihoping cough. mycobacteriusm a u n complex, meliodosis leptospsit. tetanus scarier feer .n ep e in m a roup A S0>ej4Cn m 4N disease> Stvprno u Toxic shock syndrome, mcnooocca dieae, bacruma, strep throat, Typhoid hear ty pe sahmonclloWs dysentery, colitis sahoneilosis wa h gaoenttis and enteroebti' baeillarv dysentery amebic dysentery, shigelloas, diphthern cutneous diphtheria rcespiratoi v dipht benia. L etuornaires' disease. tubere nlosiS latent tuberculosis Ftemophibius inflUenzad In txphoit fever vibrio parahacntolyueua. \ibrio vulnfleurs ibr o yersimiosis. Whipple'seae. acute A pp Bis mniwdngiAs. Ai6cphai timo impengo, el btis uar bules boil a ne, sepsis septicemia, pnumoni, mycoptasma pneumonia, meningococcal disease, mnenongt W aterhouse F il d e richn syndrome. ptomame food oisining Staph foo poison Toic sho sy drornefecroting pnumona, snteenmia -aciu ac motive endoc arihs an Afection of swcal glands ie.e [bdrdernt suppuratit1} a blAcoina dae tranaced by a tek (e g. , otky Monti Spred Fem r, [vine JAms , botuliml 'gque (e g.. AMbomC plgue, nncumornic plhgue. tuilarerr a ujellsis acute cnteritis. nonnonococcat urethitis luniphograntiloma vencrom, track homai, inehusism conittnetis vs othtie neohorn. fittaCOSiS 4 psetudommrnbranous cotis, gas La ene, acte food poitong, diarrhea tra\ tii'. diarrhea diarrhea in infants, hemnorrhaic colitis hmolyti-uremie syndrome, broxchivs heerios amacrobte cell thuts reptie ulcer, Pontiac fever, crstitjs, endometritis, ot Vs media, siunsis strentocoecal pharvngitis rheumatic fever, erysipeias puerperai: fever, neerotizmng mnistis nioocomiai inteetions pseudormonas mftection, and/o cat scratch disease. 5> Patienm Pr)ndations 90}17] In cerain enbodinets ai sNAG nanofiber compositisondescribed herein may be admisatered to a rave ubject. r a sUbect that dces not have a bacterial infectnio in one 54 unodiment a composition desert tbed berein is adniwusiered to a nave tid3Re~t that is at risk of dcquDi ing a baCteriai eCtion. 1(1011 in one embodiment, a sNAG anotiber composmton described herm raw be adionisteNd to a paint who has tesn diagnosed wth a baeral maeetio, hn another embodtiment, a. composition desenhbed heremn may be adimmrutered inoapatiemt wriho displays one or more symptomns of a b'aCterial in fectionL 10012 to certain embodiments, the compositions described herein are admhustered to pauaents diagnosed with a bacterial infection. In ertaim emnbodumnents a rpatiet is diarmosed with a bacterial infteon prior to amnstratiti of a composition descried herejFo example. the composaions deserined herein may be administered to a patiM when a bacterial attiyen is detected in a biologicad sample taken from the patient, hn one embodiment, a biolugcal sample is obtaied nom the sue or area to be treated by the eompo-tos deribed herein or an area to whieh the. coDIpositons descred nerean are to be adminstered 1n one embodrm-,tt a swab is used to collect cels or pus tiom the sie of the sutected mnreion to dte0t a baetal mketon in another embodiment a. flid is agiated from the spected site of an infction (e a wound) to detect a. baetenial imfection.n I ve another enbodimet, a Ise biopsy s performed to detect a. bacterial infection, i embodint w here the suspected site of an infection a wound, wound culture may be pe.rnmed to detect a bacteria ieetit another einbodnmnm the bol a sample i cbramed trom blood none, spuum or feces of the parent i som-e' enhodimnentN a blood or a urine tes may be Periftn-id to detet a baCtenal intenoni eC. , 'vi a bacterial intieeon issuspecJ to be a spread into the blood or other tiusu- orgaNs) In NOme embodiments the collected sample (e g, cells, tisues or fuid; i\ teted msin DN deteedon methods sach as PCR fIor presence ofone or more types ot bt..la is.n other fmbothmens, urmrnunotlureeet analysis, scroo. ctuhre (e n , blood awr culture or any other test Knon and/or practiced in the art may be used for lab toray dignosas ct'ba<tera ''ceon. 1001731 in other specie eimem the compositions deseiibed he rieay be administered to a patient diagvoso ish or dlkplayimg one or more symptoms of a disease associated wih a bacterial in trn In u ariai embodiments a paint 'J diagnosedw fit a disease associated with a bactnil ftton oi displays one or more symptoms of a disease asaci.ated with a bacterial infection prior to administration of a composition described hXZrem A disease associated wth a bacterial infection may be diagnosed by any method known to a skilled artmsn wneb m e\abwnAo of the PSaient 5m pto mS and Or detectamn of a actrua antigen in a it Io0eal samp[le of the paul eg., as deciehad aoveC In Olle ea\ pie., the conipostons dCe'cribedeei m be am mnyteled t ia pan eur d aulnosed i k disease e oied \wit a amteial ifc i by a rati g ph yeician or another nmcddi prfesiona ln1 anlother exampe a patint maiy use the composmtoin dieeribed heren upon detection of'one or moe esymptomsa of a diease associated w ith a. bacternal itecten. 100174j 1 certain mbodiments a. composition described hereis dm SMere to a pat/ca heo has h en Jiunosed with an itction. ~. the bacterial mntenon by Edt s a. (TOUM0 $ $u CN NOW ztrahotnattt. C nidop u tact1 SIMONe td/nbd nt:IV NY sd d unostnu ArteWS)(; itaM , .sly nit 6,00i, tvnc sc//o v n hay'n& to Wooghi/ A w no. ianu baut nion. L aune/ ponvlot pi p naphnr a 'pr 'terragcts I dYe~ut temo C ens itRaxc camidha zo a hashomactetn'ma is p das t o e te neserbe he the art. In one emoiet, Oconmo deseribed herein is administered0 toa patient Who has been diagnosed with the bacterial ifectionby MRS A or.Peuonoauacrtginosa s0 17 n rain embments, comrnsition escribed heei is adminiterdto paent who has been diagnosed with a disease associated with bacteria inection et leprosy O ismiN dieae cholera, anta (e q. ctngHs atrhax pSkna anthrax phi mrr a onato p setp arthi sypu conx ta syphls whoopnucough, n Cter COpe meiddosi eptspiros tetnus, scaret ver strp infection disease. baeterimsia, strep throat Typhoid fever type saimonel osis. dysentery colitis salhUalloas1 w ith gastroenteritis and emtoeoli bacidary dysentery anebde shigellosis diphtheria, cutaneous diphtheria, repsrtor\ daphtberi a, Legiomn&irea disease, tubefeulosis. atent tulberelkis Ohe1mop0i) La Ulfl n t. B, typhoid fe\ er, vbrno parah aemolyticus. \ibri o v niieius. ihbio, vetrvmous, Whiple' s diease, acute appenidietta renia s. eneeph'itisN impetigo, eihtis, earbune boil acne, sepsis enliceni, pnutmonia, vc coplasa peetrniolid, mii. lg oceal disease, 10eninits avnJtrowse Frideriebsen syndrome, ptomaine food poisoningm Staph food poisomng, OXic shock syndrome, necroiz/ng pnerntom itt septicemnla, acute inteen\vc end eartis. an 01 uctioP ot s\weat lands (e.g. lhdradenfits snpurat Sa) a hWtenid a m d a tick (cg. Roek Mutain spotted Feei, Lyrme disease). botulism. plague (e0, bubome plague. pneumonic pauue uItaremtA, brueello {Otacute enteritis nowgunococia trthnti lyphiogranutlom'i Veneriutm, trachona inclusion ean lnetiu i tithe new born, pnitaeosis pseuidoneembranous e,:ohits gas gangreneacute food poisoning diarrhea, traveler's diarrhea, diarrhea in infants lhemorhagai cohuts bely. tie~urcfmic syndrome brouchni buhaeniots, anaerobic echuhiis peptice ider *ontiac feter "vaitia endomcrtts, otits nredid, sinusats. streptococcedl pharvnelis rheunmatte tevOer, erv p'lis puerpera t vr, nearotn fasciN s TNsocomum idi int'eeons psedutoas infection, and or eat scratch diseasM 1001761 In some embodiments a composmon described berein is administered to a patiem wi h a oacteoal iehooneformanist or hetore rop t o the infection become severe lg.. , bre the patient recluires treatment or bospitaltationt bn some embodients a compOsition describ Aed h i admmistgred to t patient oth a disease fier synYtoms of the diseasem wnunf or after s mptoms of the disease become severe eg after the patient requires treatment or hospitaaiom 100177 I some embodiments u o be ad nmtrd a oms ion desnhed here is an aeima n certain embodimms the animal is a bud in certain enbodAeets the anmal i a eanein ueartain embhodnenmt, the anmmat is a feline. .1n certain crmbdihmems, the aninal is a horse. in wrbin embedtmts the animal is a com. in certain embodiments the aimal is a marmmaL e, a borsc i NWie, Ihse. or primate preferably at n uman I0 sonic enmbodSMents the auntmal is a pet or a tmo ammal isP1 ah iran certain mbodnments a stubiec to be adnunired a composition decibed bereh is a human adult, in ertain enmbodimients a sub.c to he administered a cenmpoiio desenbhed herem is a hunan aduht more than 50 years od in certain rmbodiuents, a sudiect to be administered a compnositon described herein is an elderly huma subjeer. is a premature human tiin, la certam eraberhents, a subject to be adimatered a Compouno described herein a human toddler in certam embodiments a subict to be administered a comos5itnon descrie htieein is a human cld. in cernain embodiments a subject to be administered a composition described hereinmi a huma infant in eraim embodimnents a subject to w hom a composition deserbed herem i adImmsvered is not an imnt of less than 6 months ol Yi a specitIentmoduimnt, a subject to be adimmatered dsotee heren is 2 years old Or younger, [001801 In yet other embodimems, a composition descnbed herein ma be admiistAed to a atient wh6o is at ik e p at ih nrk) of deelopi a bacteial incion Patientsbat are at high risk of devoping a bacteria inection include but dre not limited to the elderly (e . human elderly) and Immunocompimsed. hr some embodiments a .ompositon deseried herem is adinistered to a patient at risk of dev eloping, a bacterial infetin, uch as but not imied to an minnotuppressed patient (egt. as a result of cancer treatments or a traisplantaton procedure), a human child, a premature human intent, an elderly human a person with diabetes, a person dnagnosed with cancer, a patient n has been treated with a course of traditional antibioue a patent who has undergone a surgery, and or a panet wit a wound in some enodimnt the compositions described heren may be admiNistered prophylaeuiealy to patients n ho are a~t risk t'r dev eloping a bacterial infectnon, eQ gThose w ih compromised immune sastems due to, for example ag maincurilnment disease, chemotherapy , those who haw been treated with a course of'tradiona antbious s or those who have a wound {e g., an open woand} In other embodimentsy a a e t ns ot developing a bacteral i nfecun s an H Vl DS parent, a cancer patient a patin sx ho hns undergone a transplant procedure, a patient with astnma (e g- sev ere asunu~a t a drug usera patuent with a dermato logic conditions, a patients with an naive device eg an ira\ awculdr catheter) a heaIlth care worked or a paint who spends tme in a condined flWYily , a priw s 6 soil inmate, a 5oidior, a patiem i a lon term heahare faciity such as a nursing homen wt0) A patient to he adnnstered a composition described here may also be padatient utih a chronic obstruetive pulmonary disorder ((COPDU) emphyema, rhimiius bronehits, laryngghuis, tonsilltis and/or cytic fibross. rhios. ywnchvi 1001811 In cenam embodiments a compostion described herein i adomstered to a patient who ha not been diagnosed with a viral onfetion (ce.i. Hi\ \ DS u i n yeast mftecuon. in ertaim enmbodinments a composition descenbed herem is.dminitered to a P.athnt sho does not belong to one or more AO the Wolow in patient groups din immunocompronsed patient, a cancer .patin, an H1A A IDS patient a parent nith athmna, a patient w ho has udrnea trnmsplant procedure, a patient who has undergone a sureerv, and or a patient with a w ound, In one embodimentthe patient to he administered a composition described herein does no haSVe a wound {eu, a cahron wou, or an open wound due, e g to a sttrgry or battlefield trauma). 101821 in certam embodimens, a subict to he administered a composition described herein is a: ubject w ith no or low lev el of expression of one or more defeznsio peptides or a m u taion deltior in a iene or genes encodo one 0o more defensin peptides in somne embodimenms a subieet to be administered a composition descnhed here is a object with no or low or altered level of expression of one or more (dYRfensirs le. DEFA 1. D FA I B. DEFA DIFFA4 -LEAS, DEFAM one or more defenses (e. DE-B . DFB2. DUB4, DEFR I0A, DFRI104A, DER 1.08B. DUFB it7B, DFEBI0,A.FR iP0 DFB$ 12. DFR 114, DEEB 11 K DL '), 11 i DEFIBil DEE 124, DEER 125 DNB126, D L 27 DEFB1i2~K DEF821±1 DEPEBl3 DEFl B36), and/or one or more (-defensis (e. DEF'T Il) in som embodimem, a utviect to be ad mitered a composition desenhed heren is a subject with no or low or aered leel of expression of one or more of DEF 1 DEfA3 DLFA..A DFFAS. DEFB. DFMR3, DER103A, DFFi 4A, DFFR0IaB, DEFBI 1 2 DEFB 114, DEEBI1i DBLE BI 11DF. D' R \ EF1 4L D B EFB1A DE B126 DFB i 1t. DI.1 P120 and DFR131. in certain embdmews, a subjec to be administered a 6omoson des ribed herei is a subject with no or lo or adhered le elf expr on of one or mor f6l1 r receptors (eg, TLRI, TER!, TLR3, TU.RT Ti R TM, TERA TLRI. TLR T P 1 RLi and or TR 121 In yet other embodiments a subject to be adnmistered a compflont described herim is a subject w ith no or low or itered 1CVel of expreson of one or more of I L-. CACANM3 SP AG 1 bSG Ril 1k receptor IR A K , I A2, RAK4, TBRK 1, TR.AF6 and IKKi In some embodinment a sueet to be adrinstmzd a Composiion described herei is a subject with no o or atered eel of exeson one or om of lAKT&GIRR, TLR I, TLR ? TLR4. TLR7, TUt 8 'fl it) and TRAFE. A low level of expression of a.ene ia level hats lower i. .t, mrme than .25 told, V5 fd, 2 Wbhd, 2 ibod, 3 tSHd. 3.1 d fodis f olddt 5 fold, 6 fold ' Iold. h fbld.) t'okt 10 Pold lownr than the normal levee of ewpre\inn, wherein the normalis he leW of Crresion is considered no'm1M in the sPecies to which the subject bekop by a siled arsan and or toe lae o expression mi the maort of the subjects of the same species An akered le el of expression of a cane iq a e e that dif'er tege, by more than 20", 30%, 50% 75 1009. 15TQ' 200"N 250;. 3105) from the normal lev el oftexpressions here in the normal les ci of' exprcsscon is the tex el of'expression is cosidered normal m tho species to wuich th subject bemgs by a s ied artisan aNdor the level of depression in the inajorty of h Suhjects ofi te sanespccies, Wheremn the "nonnai' eapression of one or more defensmugn, is: ti the avergC e>pression *csJ known to be found in sunjett 0d a b l ol not d d e eo" Or tecton to le i ated ii) the uaverage e xpresor lev el dkteted m abe iSA e, ten., twenty, Xw~ enil ', li LI or more anubjects not displing syliptoms or not dioitynnd n ith the disease o1 in feetton to be treated; and or iii) the level of expression detected in a patent to be adnrmstored a composioon desenbed eremn before the onset of the disease or fetion. 10 0Od1$fdmimfaraatio of NAG.anotr coatiitions 1183) In ertain embodiments methods are desenibed hemrino treating or prevemnga a bacteia mfection or a disease associated with a bacteral niection, where a compositio eompnmg the sN\G nanofibers s'topicall _. . . . . . tsadimtered to a patien' m needI oeuc tr eatnt.m In sore e Od~iments. a. sAG nanoiber coMposnion la applied topically to issue or organ which has an iereased rik of a bacterial infetitn or disease' 00184 In some embodmniki effecie amount ofthe NA nmaofibes andor a sNAG nnofiber composnion is adnenstered to a suict [011 5 i some emibodm i s, a composton comprisin the SA(AG nanoafbers is admmistered topiealty to the te o4 die bacterial iection in. a patient or to the site affected by a disease associated with bacterial iNfeeon in yet other onodaert, a sompositon comprisig the sNAG nanotibers is adumstered itopeaLly to te st and around th sC of the tacterai rfection ml a patient or to the site atWte by. a disease associated wit hA ctirial fnn. In vet other emndiment. a composiOton ompAniis 1 NAG nanofibers is applied in proximty to the site oftha bacterial mttion in a patient or i proximity to the site aeted ny a disease 60 atsSOwv~ With baCterial infCeton. In yet another emnbodiment. a composition Coinprisitg the NAG nanoihers is adminstered towcaly to the site at high riSko a bacterial infection, 190il The sN AG nanotiber compositIons described herein may be administered by anyc ot the manly sitabe means oftopieai administration which are well known to those skilled in the art. inehiding but not limited to topically to the skin, topically to any other surface of the body (.g,mucosa surface by inhalation, intranasaily. vaginally. reCtally. buccally, or subliigualy, The mode of topical administration may vary depending upon the disease to be treated o; rvented. The sNAG nanofiber composions can be forulated for the various types of topical adnn nta 00 in one embodiment a COmpoiotin comprising sNAG taofibersis atpied tote skin o a patn For example; such eompositan may be applied topicl. to the skin of a paint for treadn;g and/or prertdg a baeteriiiinfection ofthe kin or a dease of the sn that i associated with a bacterial infecion In another embodiment a co.posiin described beremax he apphed topicahy to a ucosal sur face of a patient. For example, such composition may be applied topiedily to oral inmeosa ior treating andyor pne einting a bacterial winfeton of the rmoutb or umas or a disease of the mouth or g ums thdat is associdtod with a bacterial infection. 9O919| In some embodiments, a composmon comtprinu sN AOOnrtv'rs iK appli to the w ound in a patient F'or example, sucecomposition may no applied topicala o tiwctl to site of the w ound orin proximity to the site of the w ound ofa daie.n tfor treating. and or preveng a bacten~al intecton of the woumd or a disease associated u mh a bacierial infection ol the wound. in oe suCh embodiment, the wound is a bacterially infected wound'. fot dmpie, as diagnosed Sone of the mthtods describe herein, The wound may be any one of th rtpes o' wounds descnbed berem ihn v other embodiments a composnm conpris s\ m nanotib er s a not apph1ed to a wond i a patit:, or is not applied to a bacterall infected wound mapatiemth "nI n I some enbod intents a cnmosmition chernbed heren may be applied topially to a senitld urid or anal sur face/area of a patient For e sample, atiet compoattion may be applied topically to geital, uria or anal surface aea for treatmg and or prescating gengiL urinal or anlal bacternIn mtection or a disease of suCh tisMB\5 thatu a oaat~ded wm abheriad!infeton. 9I ~ The above-listed methods for topical adnm Istratim may 'n;o tie adtm tstrat of the sNAG nanothCYr in the forn of a Cream an ointmenL a tel a liquid solution, a membntne, a Pint. 61-h r"ay olr : a spray, a paste, a pOwude or any other Ornmdatlm described heremn or known in the art T: sNAi nanofiber ay aiso be applied in ar reino a. bandag. tOr examptoe tn r Tocled mteetionsvcnditions on the sikin ot a parent 1001921 in sonic embodiment a composton desnbed berei may be applied as a spray into the oal cavity ando1 respiratory system ofa patient. For example, such compositon may be apphdo as a spray fr oreaing adir pievanting bacteral Wifction of he mouth, nose, gums throa or umge or a diseaseicondition of the mouth, nose. gums throat or lungs that is associated niith a bactenal oteetnt in one such ermbhmcem the composition may be tornihted to be adrimm Ster ed as an inhaler. f001931 in some embodiments a eomposbton dCscribed herein may be ippied as a suppository in the rectum, vagna or urethra of a patient For example suh composition may be apphied as a suppository for treating andor prev enung lsatei al ifnun of the digestive tract, urinary traet or reproducti\ve tract or a disease of \ud wbissue~s thdt 1\is \Oo.atd J with a bacterial in fecbon. IX9194| In another enabodimrent, a QOmfpositan de< ebd neremn may ue applied at the site of a sargical procedure. For exarple such compositionm ay be sprayd, dppnied as a cream, ointment, gel, membrane, or pon der, or coated on the surface of the vtue or organ to be subjected to a surgical procedure or that has been subjected to the surgueA cirocedure. In one embodiment a comnpostilon. deseal\'d here in is app lied at the site ot the surtical incision, at the site of the ecsed issue, or ax the site of' surgical stitches or sutures Such administration of a compositiom described herein may prevent a post-.surgicni intleetion. For example, a comr~posittn described herein may he used during or attr a surgical procedure w hich is know n to pose higih risk of: a bacterial infection. Sxurg ial procedures that are known to pose high risk of a b ata infection i1relnade biowel resection, gastroiniestinaI surgical procedures, kidney surgery, etct A composiion described herein may be applied at the site of any of the ahb\ e isted or other surcalpNcedure 1001951 in yet other embodiments a composition described herein ay be coated on a device for example an oral hygiene prodnet, a catheter, a surcsal nornment or another product to be ued n or mnerted ito a paent, in order to present a bacteri a 1 ofuin in a parent. (09P1 NIn sonme errbodircents methods c ontempnlated here t. lude a step that includes detection'diaenosis of a. bacterial infection mn a patient. in som mbodiments. ma o~ f'e ,su~a roeun hc n nw o oeQ dectiton'd n siWnles aest or asay Ato one OTe acea. or iacterid uniiens i a bioogical ample of the patient n other embodimentsdiagnosis invohves assessing we their the patnt has cne o more n ptos oa acter NKetioo dOeasessotated wh abateri infection. j00191 The composittons described herein may ehibt sustamed release pronenies and/or may be adminstered in a tormulatio resulting in a sustaied release of suehc tiot1Um I) On embOd mats the SMN anofibewr bioderade over time as desrieod in Seeion 1, p arnd these properties of NAG nanotibers may lead to or contribue to sustamed relase of the conmpotions described herein i yet other embodtmemt the cnmpostis described herein arc formuted to display sustaied release eapbities usme am methods klnw inm the art Ihe compositsons desenbede h eim may exhibit sutaincd relese over a time period equal to or more than at 6 'hous, 12 hours. l house. 24 hours (I do)ne 2 days, 3 days, 5 days, 7 day v eiekl 10 days 14 das ( weeks). 3 weck or i weeks ;-ter administration of the composmon to the patient ( O1991 C ontrplated treatment regimes include a single dose ou aI Ynl application oF a sNAG nanoiber composition (eg. et'a cream, a membrane or a dress Q d or a regiment o' multiple doses or mutipe appliations of a sNAG nanoiber composition dose or an applkcanon may be admiered Wornly, daiy, weekly or month. For example, a dose of s\ \(nanoie composition bas admahtured once a da tce a a, three a A, lout times a fivdye times a da cvery 3 hours. every 6 hours every 12 hours ery 4 bo SXer\ 4t hours, every 72 hurs once a neek, 2 times a week. 3 times a wek, very other da, once in 2 wecks once in 3 weeks once in 4 Wks or once a month, 001991 A sNAG anofiber composkion may he admiiisered bth a duration eqL to or greeter than I weeL 2. weks, 3 ek I mn'th, 2 it v rienth. A month 5 months 6 months 9 months I ,ear, 1.5 y ears ' s ears, 'z$ years 3 years 4 years C years . yeats 10 w eas or more, I one such wnibodinient. a At3 dnon her rZtpostion does not cause any \ide effects or causes ony mdd side effects (Iurng the duratin of the treatment In one sudh embd'imenm, a sA nanotiber composite does not lose its effetivress or does not cuse generanon ot reastnunt sotin of baetria in response to the treatment, In another enthodonn, a NAG nanoitber composition does riot cause irritation (e g, moderate 0' sevr e irr at On or allergy (e g., moderate or eer e allergy).

1021 C onentration of the sN AG nanofiber in a composition may vary. in general an effective amount of the sNAG nanofiber i used. A efectiWe anomi may be an amoutt sufficient to achieve one or more ot the effects described herein, for example an amount cufective to reduce or eradicate a bacteral infecton, or reduce or eradicate one or more symptoms of a bacterial infetion, For example, a composition may comprise about 0.2 to 20 mg/cm 2 of the sNAG nanoibhers per dose/appication of the compostion in a torn suitbr topical dellecry to a patient in certain embodiments, a conmosition described herein comprises about 2.5 to 20 mg/e0 about 0,5 to 20 mg/cmn abow 1 to 20 mupcmn about 1 to 15 g/ct about to 12 mg/vcmn bouti 1 to rrn". about I to 8 mg/crn about I to 5 ug/cm about 2 to 8m or about 2 to 6 ng/em of the sNAG nanoflbers per dos/vppiiCation of the composition in a farm itdablefobr topical delivery to a parent 17 Ymbn al ~~ U11 1002011 The s\AG nanonbor composition may be adminatered in conjunctIon with other thermies such as subnomeen that bow h nn stem, antbacterial agents e, ,:m an antibiotiet defern pe tides, defen Ic peptides, pain reief'therapy pe gan analgesel el'er rohetetherap, aid'or ohcr nsr or dgs known to be eft e vgaast or commonly lised for tlrt'atm'eul atfd'ur pha enin)mof ol KR tei al mnteettls or dZhAt:Lt \ av4Ocetated w\ith bueital intections. [0P2021 1n. some embodimnts, cm ithms desired herein is AdnmWred in cosynjont with an additional antwbacnal naient, for example an anibiotic. In one sub etbodunent a compotin desrie od herein may 00 U5ed to creator 4 disease a4saciaed wih a hael infWetion in coninnoijoil ith a standard therapy coonly used to> tr eat 8ch bACteral int'cto or such diocese. In one etnbud'mrnt a composition described herein may be ad m nttred to a parent diagnosed with or display! ig vymptois of a hacterta ndfecton or a disease associated with a bacterial infection in coonctIon with a standard a.t-bacteria agent (e., an antibtore) known to be ectivo against such aScteria infections or seh disease. 1002031 In certain embodiments, a composition desired herein is admimstered in conj unction with an antiiotne ofrone of the iollou ing classes ot antihtics milcroh des igg., ery thromnein, aivbrormc en'x aminogly cosides z amikacingentanrucn mnueomycein, tiuimqtmnolones ( er. dsprofioxacio,. lavufloxa in), penicithms (e. p. pemucillh, anopicelnn, t64 amnoxu ihnt v etruev cns ( e~g., tetr aeyeme, dioxychn ta :. ad car bapencems (e~g. -mropnem, imipenemt. In some embodiments a compostion desenhbed herein is administered in conjunction with an agent ( e g ,an antibiotic) efi'ectivec to treat or prevent or commnonlv used to treat or prevent an 5. awrs ofteetion, \1RSA \ meet ton. a Pt encmas iofeuon. or a C. dAdh/k infectiot, 100204 In a specific embodiment, a compositim descnbed herem I admistered i con junction with one or more of uancomvein, sulfs drug esg., eon'noxa zoie inmetboptim sulfarmehoaaok i. terig eine (eg. doxehme, rnnoevehneY. efnuamyem, ozobdnose (e.g.. linerolid}, daptonmvcin, teicoplanm, optempnstm /dalforpriso:n (synecid& tigesy yinec, alicin. nraei nitrofiurantoim, hyddrgen peroxide, novobioenq ndImim, nethy glyxal and bee detensm. A. composition described heren may So be admimstered in conjncton with a dressing comriprislii o'ne or miore of hy droge'n per oxde, tohramnyem, ch 0rbLidine daghcoriate chltorhexidmie gluconate, 1ev oftlucon and. ahr, ore embodinxe a comrosit on described "clein is a raistered 'ith oae or rmot of the Mlted agents to treat oi preleu a ;t r infection, and paretwCulran MRdA wietion (60UO5I In. somn ' mrboditnonts the comlposltiOns descrhed herent are ahmrnnistered ber'ore (e.. 1 mmnue, 1 minutes. 30 mmteS I hour 2 hous I hours 6 honrs 12 hours 24 hours or more before, or any' tme period wn betxveen), simltaneously with, or alter ie g., I mmte, 15 Aruutes, 30 ndns 1 hour. 2 bois. 3 hows. 6 hours, 12 hos 24 hours or more afmer, or an ti me period in between) adalinistr ation of another therat\. ['or a exantple. kuceh Composttions may be admimntered b 'for, siinuhtaneuushv ua th or aftkr admtinstrmnon of an umn-bacterial at em (e g.Q an anmtbmotte) |00206| in some of thee eambodwmenth the composieon desribed herin nay be adnnamstered to a Paut to treat or prevent a bateri infecin or a shaease associated wth. bacerydl section 'ter tWle Wannt has Wndergone a course of tremmiel of the bacteril incion ith another an-tie al gert (' g , an antibiotic in sonic embeduns the compositions dsci "d heremn 'd eJrnin ~ered to a patient whso has dev eloped resistance to one or more antidbacterial agents (e g.. an antibiow I in one entbodinment, the composit ions described heeJn nmv be administer ed to a patnmt >t he has undergo :a course of treatment wvuh an anubiotie (e g , an antibiotic standardly used {or the treautment of such haictenia iJQuimt) andI developed resisdance to suc h antibiotic. 6or f00207 }-loever, m ertam eCnOtOKifmnts, a sNING anotiIr enmP0iflO On is adntnistered ,lone. In ont' iuch umbodi ment, u NAG nanotiber Composition is not administered vwih any other therapies, for example, it 1s not dtnistered w ith an inunomodulator, an antibaceinal gentat.g , a antihtiect sodefen teue aP defensmnlike popude, a pam relief Tnerapy ic iman alesiet or a felver relief therapy. n one embodiment, a sNAG nanohiber compositon is not admillnstered im conjunction w iTh an aniDbiotuc, In certain embodanents sA% nttnotihei compositionls arc nor. admonistered1 in conjtinction w ith tin antisriralI aemt. an anti-fungal angent or an antrifyecast agent iKit I9%9i)2O A plarnaccticat pack o0 kit whbei eornpr isc any or the above iescribed AN AG compositions iS also contemplated. T he pack or kit may comprise one or more conmatners tilled w ith one ofmnaingreduents cotnnrisum the c 'p911U d ue 1d hutrew.. 'T he nttion. is or a Slooo Mr. tosmi neri'~ reterabiv contained wihin a sealed, Water proof sterile pmkge which facilitates nemolal of the composition v thow contanurullen \ateris trom which containers ray be mnade onude aluminum fAil plastic or another conventional material that is easly steribied. The kit can obtain material for a smgle admimnnstrainoorO mn le admn nsn s of the C omPomtni JflUli\ ab \ wherein the. maial t or each dminirauton is aro\vided ina sepdC.ate, deptui \flen c package, [002091 10. another embodiment. ai contairir hais og~ dal compartnments is provided. A first compartment contaims any of the abon.dcscrthed a A( composnios while the second orpdrinent contaiso th1er at1ve agent such das ana er atlbtsC ri al agent. In the tleed or the ehme, c the Cenpositiotn in the firtecompal~trmem t:an ne read lv cormbmieil with the a gent in the scond. compartment fir subsen*ct adinintstradtin to ,a patient O02O Addtionaly a kit designed tfr emerenco tu md; Use can also conta. disposable pre-steri lized instruments such as scissors ,sea Ipe l clamp, tourniquet, elastic or inelastic bdages, or th fi01 f9X1 I Optional\ associated w ith such kit or pack :can be a notice in the torn nresCrined b\ a pt vernmmeUla agency repulating the mnlfirGt or sale* of pharmaceu~ticals or biol ogical produicts, whblch notic c 1 elets approval by the agency of manuteture, use or sale Fir human adr0instr'Ao n ir '*\ample, a kit gan comprise a n vergrdg FDA pro al candor intsions for U1e.

100212 The kna enmassed }ein AS bin ca s n e application ad thosn d 63 Exmni 1:NAG Nanufiers From a Marine Diam Pronote Wond feanan nd ou E ssin i n AkU N t 64emnen Pathwa 12131 This example demonstres that sAG nia hes pomoe ctaneous wound heaiinu and cyresvion of defensins, and that the Akt 1 -F pathway plays a contralrei the de~ien of cutneous ound healig by sNAG maofiers 6.11 J ateniaantI Mta Iethods 100H4| sNAG.Talidrn aofibers are produced and supied by Marine Polyner l'bnolonies and formed ino suitable patches for wound treatment Widtypr Black and .Ati null rmce were housed at the \MedIal Lniversiny oftuth Caroina animal t'NWilies, dt\pe and. Alt i null tcee, ages raning from eight to i2 weCe were anestetzed wlh 50% pure oxygen and 10' sofurane gas Irnmediately berbre wounding Nair Hair Removal Leben was appbmd to hei dorsun to remove any unwanted hair A dorsal 4n u il eular area oi skin w as removed using m excision biopsy nunch Tahdern w as placed onto ec wound at day 0 or WAnd isure k ft Atuedk. At days AS and 7 the wounds were photoraphed, measured, and excisd using an t nan bi&psy pulle to measure complete removal of tie w found and :SrtiOUfi skin. vldtvne and Akt null wounds with and without Iahder trnent were embedded in paraffin n preparation for .&E and innunnfluoveseeni \tai 102Sj Paaffin-~embedded sections were sectAned and placed on rieroscope slides ter airing Sldes were washed with vlenmes to remoe paratin and rehydrated through a serin of raded alcohols. I hK seulons were then incubated u 0 t i rton t .00 for per1Ambizatuon, Secti n. were incubai d in a boiling Antigen R eirv a huion 1 Animal sum u a\ used q1'r Noekng before meubating in the primary got antiods Aydcfen 3 1:400 lution, he sections were then mcted mci the primary inttbody. ox-r Arght t in a hurmdity chamberAn imnmanfiuoescent secondary Don key ti-goat 4M3 a nibo dv 1:200 dilution w as used, followed by nueaar staining with IOPROG3 nages n captured using confoeal nicrocopy [002161 Hmatoxyl and conn staimn w as used to visale hasle sutues such as th< epidermis derninuscle, and blood vessel and to determine the orientalon and apromate location in the wound. HA. Ftaing was als used We begm to identify which ell types are atimutated by Taiderm in an Akt neendent manner 67 100217j Oher rmaterias and enthods arc descri bd infire desrptions a in de result secton below. and perfoned in accordance withie methods known inthe at 6.1.2 IlesU ItIs 010021 j sN A G Kanoihbers stimJuate Akt I action n. tn upsrcam regulator of ET I.Figu re AA shows a Western blo analsis of phospho- Alt in response to NAG and sNAG stimulation of serum starved EC Figure 1 I shows RTPCR analysis of EC in feted either with screamed control or Ak tI shRNA leiviruses and assessed for expression of Ets and S26 as a loading controL Fgure IC i sae a Oig transduction pathwav transdncing a sinal from sNAG Janotibe to AkiA E nsi an Defe piS 00112 191 . led wgod tealhin -OI k aninia ais praI w i rwuo di ?2ia 3d l mAGe) eawimn Figure 2A shows represent ve imagesofwoded WT and Ain a mice with and wtthout neatnent of la hderm. figure 2B shows IA&f. stairng of reprevema ae mouse skin secton\ from day 3 wounds. H& signing of widtypc and Ak . wound exzewons idiext a. Taliden depden ddent increase m keuunocyte proh Qroi and Wrgraton The dashed lines indicate there ofg p t cross de unund marm, in both the wdtype and At I treated pounds there is an an evdent nrease in reepihehianu rn the nound nyrmdn compared to the Wtldtpe and A! I control 'i hmdicates that Talidr mreases kertaimoete eerwtmnen inderdent of the Alkt pathuay. Althouh Tainir mduce omtlete :reepthhzation of the epidenms aron the wound margin. hr i subtanial lack of rev aseul ariation in the ndrygtasue compared to the wlctvtc I a 'l h i dent by substantial hem rrhaginn and intidtraon of red bood cells m he AktI anmnale 100229\ 0 \ itr G Ian w nunuak ctoane. and JdenAt wqvresuoa: in prmarenothUasA cy/is Figure 3A shows. imnunuhitechernisty of EC treed with or bot sNAG using a anUbody directed against udetnfin. figure 3 presents L1A showimy tha nanobber treatmca m QC rnults in the 'erto s defensins 1-3 |10221| s\X4 no/lo s amuat dtnm prSmn pWt y' 04 lotihR cw/h W an Alit! dlependen manner, Figures 4A and 4U show quvni taime RT ,PCRK aalvses of sCrum strained EC teted with or wihout sNAG, with or withoik PD%0 M A PK minibadrt Wortmanno (P1 K inhibitor) or ineted wih a scrambled control or zkt i RN: ie'th iruses and assessed Or expreSSion o he gnes indicated 68 00 22 2 1~ twtmuQUISt snghtt /lcesin ' tOUCtYIWVP$ iu A shows imminoflorescent stamn wih l-defensm 3 and hwolucrin annibod om'parafin embedded mouSe eutmn vos omund sectons from WT and Ad I null animals on Da x A culdnetus w ounld han model w as developed in bothl Wl andl Ai I yud "Am'ce 'o usess the effects of Tahder :n mo Ese tindsshow tha fdefensm 3 expression me eases mn l ihdertn treated dnl0Ild\ in an r\t I dependent mlariner. ihe ability of fahdernU to increase defense expression in a heading aOund has m1portmt implication for treatmay and controlbng wound derton MFiure 5B shoun qutification of ldinsi iimtnofluoresceet stainn 101 INTiinaggl ot1\ S aw foure f C sho\s hu Olnmnollorest siring of \\ and A ktl n treated and Owredted keadtinocates wvidh (iefensin 3 and ,OPRO. Noice the increase in green Defensm 3 stainig in W and UKI Tahderm treated wounds The immniotluorsAcent labein of' ound seci ons iustrates that TA idern treated wounds show an. meease in deensin 3 expression in an Ali dependent manner Although the Att treated wounds show a reaonable incease in defnos the w dutype treated wounds diustrate a moe reia ab merease Ibis indiates that rddensin 3 expresion is not only increased by application of the nanotibner, ei is at least parnaly dependent on ie AKtI pathw ay. Pieensin 3 expression seems 1i0ted to the keratinctes mdieaimg thn e\prevson is ceratmocyte speife |o0223| 4/a dependet trsra2/pt ?ar wr mnwdnir tes. Figure 6 shows- schematic of Aktdi dependent transcrption faetor binding h iS N (Genomntix sotwa 50 lop uystream of te transernon star sie was analyzed forconseved sites on the RNA of DIEL 4 and , 43 (oncc'sions f002241 The provided data show tu 2N ' nanotib sinulaion of I ua Iesuts rom the activation of Al b these nanofiberm Nanober treatment resulted n marked increases in the expression of genes mnvols ed in celulai rectramnrt, such as IL -I (a knuow nl'sl target I GE' and several defensms i Dat, ad, and s omall anti-nucrobial ne \ cndy shoun to act a bemrnoatractats Boh pharmacological inhibitin of Nh P13 FAkW pathway and AMi knockdow n usng 4h R \s resulted in decr eased @ex prsso Ofl1 th;e.e chernotactic fuctor s Akut null mice exhib-ted a delayed ound healing phenosyvpe that in partally rescund by lahderm nanoabers. Taderm tted wounds also showed an incieus in deftsin expression that is Akit dependent. 69 100225 The mrease of detean 3 expreIon ad heraimOCYe prOlerCo n Tahderm reaed wounds demonstrates the beueni al use of Taliderm as an effecmie wound neahn produce. 1aiderm acts to increase .mu-nerobal pepde expressing kermamocytes in an Att I dependent manier N esuy ing the essenta tl Ale of Ak, I in the funcion Mf sNAI nanofibers. This correlates wih the resuls rorm other studies in the laboratory (Buff \ise-Heimeneks, unpubbshed) that mhibition of'the P1 \ i pathwy and Aktl knockdio a usi; shk\As results in decreased cxpression of these ehemnots t feor t2 \hugh te m nr eased expresum 0s odfnsr is k u1-derendent U&L' stainmng ot8 tinm weut i scisions (Egure 2R) mnteatcJ .ha Taier acts mdX~eUdem of Ak t in found repjrithchzaion. lien though the new kecratimoctes span the ent re wound margin,. de underlying isue did not demonstrate the same stimulation in vascular growth, Ihis indicates the that abwsee of Akt is responsb ic MY leaiy h ood vessel and the large amut of tioatng red blood cells in the dermis. Ths suggests that faliderm is dependent on the Akt I pathway for an increase in vaeulai ianOn. 1002271 In su0)mar il SNAG vanofibers i ilAderm) mer e wound h ealing. in par* by stunm tnng angiogcnen (ii sA AG nanofi bars treatmnt of enOtelial edl aCe an Akti I dependent pathwya leading to changes in cell motilty and cytokine secretim: in) Talidern treated wounds show meresed exprsson of defendi 3 in an ART I dependent mam.r, (lvi treatment of Akt1 Unil animals with Talidrr. paiall\ reues the phienot\pe leading o markedly in creased kerimoeyte pro operation nugraion, and n bioIormancs analysis\ iricams that F'TS I is likely an Axe mI jihk 'AG activated r'abnwy leading to increased wound bcah r and cytokme seentior 1002281 al n ctvgehr these fidogs sggei a cenirdi role of the i<t1-Fi. pathw ni mhe rcspilat ion of CL nancous v4011nd healin UN AN MA natn ihers anld .5tpport tie use or thesc nanotbers ,s a noVe and effecti n mthod ror enhancing w found healmg. Wound cl.osure, and1 Um nr Inirect flefensin-Denden I Anti4macterin Effect. 10O22| This example demonstrates that W AG iA nnotbers have a potent antibaeciai effect against Saphr>ocwt waen in vo, which i indirect aO d defensm-dependent Th example also shot vh~a 1t3 sAG nanobeers induce ex press0n of detensiUn~ i. Vtvo in keraioestes and 7(1 ndothelal cens and inkvi n tanos udsdi an AkP dependentinunnt and iireise the kinecaof wound C1 Sureh tl. Materias nd Metids 102301 /),ua (Jul wrc PhmamlVa Mili~nL:LL i--I unman armbdical cord vein F( (Lona) w ere maintaNed at 32 with % C2 i endotheha basal medium 2 (Lonza) Sndothebal Sal medium 2 (FB1M21 w as suppkemented with Bc' grow th m ed iin 2 SingleQuo a des ibd by Lona procedures and mY pentedim/streptomywin (Invitrogen) Serum start on w as perornned at S049 o% co aHencvy in FiM supplmeed with 11 % fetal calf serum (Val ley Biomebt al) f N hours ulwed by nmato hi trified pQleNAc (50pNgI) nanotibns {A\G) i stn(te water (QWas ided by Marine W'Oly.r eehnologe iN Dauners Mas l'SA) The nlcNw ditomdernmcd nanaibers used in thistudy are bor biodc LradabletJlbers denA d rorn a longer Rlm \ \GI, and have an average Weh of 4Qu.i mnd a polymer molecular weight of approsimaa tel 000 . For hbiion using PM09059 5P ) or w rrunmnn (1a W K el, s wc! [ rN p rea e tor 25 mni pror to 3 hour stimaon with sNlAG (50pn mi | 09231) ~Saus ei MUsn l'a qruantutrt e experunen as performed at east m tiplhate at leasi ihee independent tdoes A l statistal ana Laos were perroned using \leroson Laell to calculate means, standard vastion and student t-est [00321 ll .t:&fl //l tr0n \4 isawon diR\A Icntwimr1al construct directed against A4 w iuere imrchased frorn Sian,\ldrich. A\ seramld oLKO.i shR NA v ector w as purchased from Addge ne - Lenreinwe w Nete propagated in ?NST cells maintained in DGMl'M eupplemrented a above. L entiuaI production uas pertorrned umpPAX2 and pMJD~CG pac'kaniro ter purchased from Adrdene using the protocol for produemg ientital ar from Addgene For infection of targi cell ~ 5 \ It 7:ls w ete plated on 100 mm plt a ahow'd to icubate overmght. The nem day ,e ere transduced using a nal concentraton of 1 pgmo' poly brene amd cit her scrambled c ontrol or AkilI shR\ A 1entivmses. A30er transduction, endothelha! cells were secarn starved overnight and estimated w Aith NAl SOft/nil) for 3 hours. Al ifectins svere mIII toed for aproprute knock dotw n by REP('R 1002331 RT!PPR' For scaquantaxive R ' \ A uas extracted with RNsoI (Teitcst. Inc. ttiolow ing nanufacture s instrukonns. (DNA wa ynthesied froin 2 mu total R MNih Suprcrlipt F's trndfand S'nthesis K0i (in liiogenl. uimg OliputodT) following the 71 igmdauiit rer' isutingi PCPR rein ,eita nnddygua~ nmoudst ofDNA and & 3 1 rho propnate primer pairt Nigm Prolino, to L ousM SsA).Al primer seqiences usedaiu Akfl F 5'AOACCCTCAGCCACACTCTOG3 Aiktl R WATGAGCOACGTGGiCTATTOT 3 p DYiia F 33 GTG{UGGTGAAGCUAGCAO 3 Defensind R P15TTTCTTTCTTCGCACATT 3 '.dtasii n 3 CACTCCAOCCAAGACTOA'T3 DfI RX Unsir 53' TCCCTGGCATGCAGGITCT %26 F A CCGCTC TCCCTCCAAC 3G S2 RJ C C G\AAT ACCGTCTTC A 3 [002141 Cyeng condions nwere; 941 for 5 rimn 3%- e o 9C 6rin, 6 ed on prnmer T y tr I mm. 72 7 ' 1or 7 n d cooled to MCY ( ye nuner wa emnirically determined to be Withm the licar range of tha y for each prmer pair used All somun iandta iye PCR n a p formed wat the nooomas protein suitnt 82. prn as iOendl controls Products s r isualired on a BioRad \lui imoagng N stem (Hereulos, CA, USA) Rea thm'ne. P(R a performed uing a di ilh CYIBR green QPCR kit in combination w i an NI 000P Reaim:1ne PCR stem bodh poehased foAm Stratageune. Priners detecting the aibosomaP \uburml S26 w ere used us ime~rnAl ontr"ol> 1002351 2reune' ho dM (Awn! aog W K \ Wild Tyx cS-iM and Ak% -!- 43 were used i all experiments The ALitl null snimals were created usM an insertional strategy at the tram s ' star site that block - \ex iaor the entire Propin woUnmg was p*rformed on anesthetized :adt mtnale imce betw een 84 2 w eeks old. Two fll i thcknens cutuneous wounds were heated using a Imm Lopsy punch (\ ilte: n to create two eemical wounds on each flank. \be were aneshenzed using aru O0: Isdo an sr 1<aporiang anestib'sta mxachme (VetEquip, hne.> Isoiurane w as used at tor induction: 2 fr surfer>' P rior to urgery hair w as remCed by depilation and the area w as w astied and 0k nhwd using Y~0 ethanor. Wounds wore etther treated ;ith A sAG mornbrane moistened wth d'tled w water or lf untreated, On day 3 and 5 animals wore authaiied and ete nOmldS were harvested including the surnding skm asm en 8 n biopsyv punch NWte) Wounds were facod in paraformaidehydc vrnight at 4% onmbedded in paraftin, and secionted to'r analysis 72 1002361 Hemamvihn and EOn S tiin &H& E9 & staining w perfmed in the Histology C'ore Facility at the Medical University of Soth Carol ina, Department of Regenerative Medicine and Cedl Biology. Briefly sectons were cleared in xylene, rehydnaded through a series ofgraded alcohols, placed in HIenatoxy in f bylowed by acid acohoL. Samples were then naced in anmonia water rinsed in etano and e to Fosin before dehydrating through graded aleonols and hearing in xylene, Sections were mounted using CvtosealXY L (Richa rd-A lan Scientifle). H4&F: sections were visualized using an Olympus B1X40 microscope (4x objective cns, 0 13) and captured using an Olympus Camera (NModel DP25 and DP2BSW acquisition SOP Wwc, 1002371 Lae&a if nin, ' . we Swidv C w bit (Quaniot(on: Male mice between -12 weeks w r ounded o descrbed above. Singi colonies of Stallm aanxaras ATRC( 253) t nre pice and cohanred ov ernigh a~t 37* and adj usted to an absorbane of OD c 053. One o f S, awu va spun at 10,00Mrpm re-suspended in sterile P BS, and 19l was used to mnoeniaie each wound. KNAG membranes were appled to the treated group tAty minute post moulaton Mice w ere euthanized on day 3 and 5 post wounding d wounds were harvested sng an 8 mm biopsy punch One wound per animal was fixed overnight in 4prabmaddehs de t 4 C and the other wound was uiltured and platd on. LB media without aoubiotic Sr material quantitation (see below). Wounds for tissue gran sutning w ere embedded mn parHfn and sectioned Sections were leen inyn an ehydrated through a senes of alcohol and were taed using a tisue granstain (Sigma Al ndrieb bw procedures desert i hd by ie ane ter I00 F'or eituring, wound Seton\ were plae in 0,nmI bacterial media an incubated for 30 fmu at 37C while shaking Clon forrmin untts l F were uamitated using a dilution ws plated overnight at 3'C' \be or colomeN per plate/per diluion were counted and (. I. in ere caieulated, 1002391 T dtnmie ('FUnm from NG treated bateria unres, S. auras cultures in solution nr treated u iti trying concentrations ofaAG 0ul and 601 _1100 nmg mn8 sNG Ai for three hour ' i lures weare then plated as ernight ait 37 and CF1 mtI were dcetermmcd. 190240 do Jena ' Peri/ 'pndyIc~ati: Three test conicentraiot (I.(g\L] 25pM, ,0 1 .M) of biologically active human 'dcknm 3 pepde Penpde rinstituet inQ wnere tetaid for their effect on bacterial growth in the infected wound beahn. model described above, 1ac '73 cocentraton nreradc V affected bacterialrwt iho sicceta o honfo layrowtb 50fa m thC T loWyt n B:iRr4t! was chosen 0: analyses. Aer each wound was ne ed whit S aureus ow peptide was applied ?er three d wonds were harested, embedded O Secong andra staring. on aved for E041 qantitation as described above s00241| g-du r mm nt bmdl Bv LS de: \ni~d Tlpe male mic were w ounded and in eered with 1 u. of S rm a descNrbed above. Mtr moeulan on w ound was treated with 0 Jug'mL of fdefensin 3 antihody (Sata Crun w hile the other was treated w ith 0I2 u mL normal coat IgG aonrol antibody (Santa Cru N\( membranes uere apphed to al imee atder amibody treatnem on day 0. Anybody was appie ery 24 hirs Mice wre burnd on day 3 and wounds wn e harvested uoing an 8 mm biopsy punch, Wounds were fixed. oveight in 4% paratormaldehyd at ! !, embedded in paraftim sectioned, and paralyzed usng ussue gram stain I mluutniatio w as performed frnom pounds harvested on day 3 as described above, 100242| /nnm oienat A/cnreopv: Paraffin embedded tisue sections were re hydraltti through 10ln0 and a eries of graded alcohols Sections ere created with 0.01 Into\ 100 and subjected to anugen retrecna using antigen unmasking solutn (\ ector Saborawmres in a prssOne cook r for anm nd allowed to cool tinset n ee laheed wfih bdefensm A goat poiyeonal amib;dy (Santa Crun involucrin rabbIt pol vlonal antibody (Santa Cruz, and TO-PRO -odwde (Molecular Proboe) Seions were incubated in primy antibody ovrMmght at 4' and appropriate secondary onno1iuorct antibodies Inr wtroeen3 or 1 hour at mom temperature, Control sections for ech antibudy' were stained without primary anmibody. Tissue sections w ere visna lied camg an O!vmpus TIuro~iew laer sceangn contocal nireosope (Model X70) and captured atanbit temperateus ng an Olyms amer Model FV5-ZM and Fiurview 51 ac uisition software. lisueseions ere rnaed urg (l0x oil rtflrrmers1rn ens (0] ympus tnetsion 0:d) 1002431 H U\'ECS were either serum sred or treated w ith sA \G for hours i future and stained 1h anibodie directed against -defi n S 4 U5 ( [ F ), siL de (TesA Red x or TOPRO 3 (13BlueV Images were taken using inmmcunoirekect nuceoseory Cell culture defensai expression uas visuali:ed using a lS MM Oven va0bUM onocal microscope aud was captured at aNent temperature, using water as the meodni ulmg LSM 510 camera (/eis Fluor 6h\\ i 2 A objeciye4 74 1002441 Ko re n Blo/ Ano Fidohelial ,is e were sar starved prior to simulauon wth NG (5phnil for a gien Ate course, Gel Is were then lNed and subjected to easternn blot nlss Te dantmodles used tr eern~ bk 0anaL s are as (bios and8-S s0umnt ofP1K and phosphoseete \kt antbody (Cell Signasl t iechnoloies 62.2 Reutdts 6.25I Kt'.rannteseand Endothetial CtIr Express and erete thr dn When Sdmuaated Wft sN AG 1002451 This example dernonan ates that sNAG treatmem modes the expression of defensju small anti-crobud perpude that are par ' the nate imnune response. 1h246| To inssUgate the afQet Of 4 SAG ta n dofon'i) esxnretal 1i r urimarv hantm umbdicai rm ndothel cI lsn uat Nere uwd Lodotheltal gIts press both type and 4 p fenyn\ when Stirnuated with s\G. As sown n mFgure 7A endothehal calls treated w 0h sN' A G Nhtow an up-reguuIson of f-deensin and &degensin I mPN\ depression nwitin hour of stinmulaon. SM .r Up-gudaion of twieeusn 4 and hr s\ A\( threaten was also observed (daa not sh0o Y AS.>to= gene arrays cntagtlng over 25 different dekasin aenies were used to confirm tle expres n O the ttp det'nesm in psa inry endotheliaul cells and. the 1tyC densins in knlabno~e es. sNA0 stitulation of endotteliad cells was hown to incrcee the expresion spet 'tilly ofrdefenasins 4 anmd S and p-deftnsim 3.Additional) dv \NA atilnuuation oh'Anman kears~nmo"Nte tinceasded expreson flf'delnsul Ike geoet ww 'al of which are listed in TblN I I ee findnp suggest that t least thre O defensm ceuos arnd pb-defenson 3 are expressed "inc vran endomehal cells and niadntple V defensin genes are expressed in prinnuy keratinocvtes in response to sNAG stimuAtioni Tabe I: Gene array anahy da reral nmerous defensin genes regulated by G IUVEC Gen, Name Keradnoetle Gene Nam Fold Chae I tdefensia .eensn 4 oeuIn 5 A2.6 Vdef nnOB +2.55 P * nemsn 1 +2 9 3defensin2 65 f3-de(fusin 4 06 P~dersin 129 46 75 1002471 To test uhie% the sNA(dpndcn t defenm exprnsin also ocurred on the proCif levet. s\A& stimuhited endotheba! clls were subjected tO tfoi O uoreSCceC usaf antibodies directed ag&At both e and j defense As shown in Figure 7W both b defensm 3 and Q- dQenimi 5 are upregudated upon NAG stimulation mn ths elil type However sirniation of prinmy human ker atinoe(lshat) wih N AG did not cause increased eresion of, defensin but does cause an increase in the exprewion of tdefensin 3 Figure. 7C T aken together, these experiences suggest that s NAG smtdation results in an up reguaton of decusm peptides im both primary keratocptes and primary endothehal Wels, 6>22& $s JS(Griptlem a in keoin spewo Rceirt MIsl 100248 Prevously pubisted data sow that s\ A st amuation of prmary endothehal cells results i increased imtegrin actiation, t I expreson and \iAP inase activaton, (\ournakis JN\ et aL 2008, 3 Vase R ;- 15 ( 222-23 Findings position Akt! pstram of tS I in endotheil clls and in tawrmp (tavenburg. KR, et at, 2003 F ASFt 1 17(1) 27840 1 To begm to dteurinnhe sAgnaliug path way responsible Aor the expresion of defertnm, endothehl coIls wace scrum starol ed and p heated with ttharmaeolo'uco mha.nmr directed agamns P3k inyrtmanni) or MlAP s (P\0 pnir to a AG iauti Qemtative real flme Pt R aayis stows that useernm i mRNA lesl amrerv tildnmished iuer inhbib'twn of either the Pi I3Rt pathw av or the \LAP kine panth' iF mgure SA XI RPCR< analysis of [ defena.3 also shows that leVel are decreased by Whe inhhakiou of thos pathway as w l (Fgure 88) sNAG treatment ofendothehal cells fot a short mn c ourse leads to phosphorya i on oF A kI1, a standard d indicator ot' i m t i an', i iure 80)' To r. on m n that ASt is indeed required fom detenaw exprein, lentvral deineryvof shRNA dir"ete J agamst Akt I was usend. Quantitatin R -P R of serum stand endoihehal cio mfected with scrtabled (CRI) control o1 AktI shRN Ama wed wvrh sA% treatment comr ov that Aki. explres\ion is reqAired for A AG-deCpdent a-defensin expresion (Figure 8D). Smce pWdefe ns are known to bo expressed in opidcliMl cils, iciiviri delivery ofWHONAM iece against .AkI illuedm human keratmnocytes (Hatit) sNA\G treatment ot' serum starved keratinoets infected xvith scrambled (SC 1 oNtroS leads to a sgnificma increase in -defensn expression tbat is abrogated by tAktI knockd u n I gure 8I )Iltese results illustrate tdt sAG treatment at ates AKtI ina endothelial cells and strongly suggest that sNAG-dependent defensin expression reqres AI\k in both endothebia Alds and keratioiytes 76 6~~2t i A Ieaaten of' Cuteaeus ThunuI orese Dfl4nsin rxedoin 002491 To conirn the dependence of Akt f or the ear ion of defenms in v w ild ty pe <md AktI null <mimul& W ereused min e&eSonaI wound healmn model. Although most mamnmahian Icukoecytes erxpress t-dekeusims (h umuan, rabu. r at, and bamster> mouse ienseev te do not express 1defesins1, }'herabre, fdefensin eApresion in these mouse Models aS focused on. reannent ofjteaneou wounds nith a dRed tam of sAM. a bn bRkdeaMadahk mermranes for three (lays results in a stattstieali snileant increase in Vde LeNsI e sples10 in kberatinoevtes of wil type animals (Hggure 9Ai inolaci (Watt, i \ 14 mest Dermaol, 41 i Suph :SA0s s training ( red) as used to mark the ker:,lnocyte cell layers and. show that the expresion of Sdef'ni is confined , the epidermial laye. to asses if \AG~ dpndnt defenuin expr eion s dependent on Akt I a tirmiar assay w's p~eforrmed using an Au i nut. ammal mel \\nudbfrm API null mice treaed with A mea r mek show a markeiv reduced induction of 3def i 3 Sxpresin (Figurt 9A } o bette vm uli e the epidermal layers that are expresg 3ewensi 3 Figure 9$ shows a represented imane oA sNAtI treated ild type wo und harvested ow da i A Vi treatment ofctatneous wounds induced iVdefensin 3 expression ma ir ' the uprnabasal layers of skin Figure 9$> Quantitative anayses show n in Fioure 9C shows no apromnate5 MlJ mdicrease in -defenin expression in sN\AG treated w ild tynpe aninals and that Akt l is required for this irese tuk4+2 iM Troeatent Increas ihe KinScO of' i q eind Closure in WT Animals 1002501 Previous resuls have shown an increased kmetics of wounclosre m iabetic motse models in response to s\AG treatment sA\Os u w x tete for a similar affet in wd type amimals. Exeisional wounds were created in ild type anals which were eiher treated with the membrane form of sNAG or left untreated, ' e setions w ere taken at 1. 3 and 5 days post avounding ad suicted to H&E staining. A shown in Figure 0, sNsA.G treatment of wild type wounds result i complete closure. as visualized by the sohd linc at day 3 post wounding This occurs two days earber than in the control wounds. Akti nul animals display a delay in ound closure: these animals do noi. fIuly close the wond until 7 days poat wounding The delay in wound closure in the AkiD null animals (s not rescued by sNG treatment data not shon). These findings suggest that aN AG not only induces defenusm expression bur also increase wound heui kinetics in wild type mice and may be a novel and effete therapeutic. 77? 6b2l2A. sNG A a Cfjecdv Antnmngrnhie Aydiatq &Aeau 002511 De n pepdes arc known to powess anthnicrobuad properties that arc active against grm-potiR and gram-ntgatiiv hatenam reamen of'ndothoeia cellS ith sNAG mereses defens xpresson (both w and Ptype) and treatment of cutaneous wounds u hNAO dramuically increases V defensin 3 expreson V i enn the antimiembul efficacy of SN AG tmcni t bafl y ineted wounds as assecd 1904252 1'o determine itssiAi decrees bacterial load in cutaneous wounds, wild tJpe and AM null ammals were asubjeted to cutaneous wound hearing, foow ed by iition ith d W , infeed wounds were either treated with s\A4 or left untreated for 3 and 5 days post mtection. As shown by 'he issue gram snmng m F igurc-s 1I and 111, xd type anims treated w ith A AG show a inmfcant reducA ann n Aam pos t e toto in b d u pst wounding as compared wAh untreactd wounds In Comrast gram shamed bisue dern 'd from untreated wu snds in Aktl null dn taM at davs post non d' W %hkm I n aeunmtisti of neutrophI nhih stam ran positie Figure I1R), ictnng a potential lack of baarial clearnec in t'hse animals that is not rescued bt sNAG treatment These flings suggest that Ai I n ul amml lavei U tadyel m nine iradiec limsiluch is nt r'"ucd ay smh A treatment 100231 o puantitate sAG specific bacternal changes in colony forming unit{(CF ), infected wounds 'rom both v id { rnd Aktl null mie oijh. 5 'AG r d or treated wer arrested and cultured, As Sho n n Figure 11tQ at 5 days post wounding buctenal number is nuarke~diy reduced (10~old) n idtype animals treated a ah s' lAG Iow ee, although the uner of bacteria detected mn ue \ktl null animals is redumed m comarmon to wild typc, 0\G treament had a little elect on absolute bacerial number in the Akt] null animals Qt 3 day post-infection (Figure I I there Ais a sminat 1-Old decrease in CU m <NAG treatedI w ild type mice a compared to untreated controls The sNAG treated Akt I null animals show a 1t 'old decrease i CPU as compared to treated Abi null animals in general, the Akts I .ml anmians have a lower bacrial load per w ound which may be reflective of an Akt 1-dependent effect on other processes in addition to deensin depression These findings sugge that s\AG treatment results in a marked reduction in bacterial load i infected couneous wounds in wild type mice but not i At i null ice, suggesting tC poss ability that de'osnsare mediatmg the anublactenlal response. 78 1002541 To show that the a.ntbacteria effct of sNAG treatmem s no due to a direct ffet f the nnofibers on bacterial growth or on their survival S, unreus bacterial cultures were treated in souion with different amouns of aN AG, for 3 hours and colony forming units were determined, As shown in Figure I E sNAG treatment had no direct effect on the growth of S &Wreus, indicating that sNAG is not directly inhibting bacteria! growth and may then be working via the uMUCKMIiatden of defensins. 632A6 A pplication el Dejeasin Peptide .ksier shAy G lndbaauniai ffth 109255f To deterne w whether addton of deesen pertde can block bacterial mn'ctionm similarly to that snon fr sAG treatment, wild type mice were wounded and mocuated wih J! a urn as denRbed abm ho and then treated w ih ti o iall ac tie hunan dini neptds (I 0pm) for three days Tissue biopsies w are stained using i tissue ram stam and CU Was inanptate& Ngare I i F-G show the rests o thee expeoeimms. infected mice treated a ith |-defertsin 3 peptide hive a decreased bacterial ioad, an approximae / fold decrease n viable bacteria (Figure II)Q similar to that shown in ild type ine ,ra ted wib ss\ 100256J One of the mechanisms by which defend expresson is induced though sTmilanon bv bacterial UP> possibly through the activaton of TOl hke reteptors delsted. Mil' and A I. uelette, 2005. Nat munoi. 161551k.) To test v eth r acteral mtetion alone is able to induce 'deLnso e\presinn within the time priods teste expression oft', defense wa messed in infecte d wond rom U wild type aitnmais attr bice days post wounds, As show n Figure 12A. b acIen inf'ecton alone does, not nduce the expression o Jefbnsint within 3 da~ ofl inetnn, as s Nhouf wi th s\AG tremn i r d tb !ype anmmnal s \ tC reatmnen of infected wounds causes appronmateck to 5-Ibld iscrcase it the depression OA edefensin within a similar time period (Figure 12. 'These findings suggest that N'AG teatnnm r avidly induce~s the exsion of defensim expression vesiultow in marked bacterial clearance in 5. aun infected w ounds 6.227 Antibodies 0iretted Akaint /P-lcfrtin t KHiatt tie Anribawteria/ Ert* A 3uCG 100257 Since defensins are secreted protein, the in yeors hypothesized that antibodies directed against V deftnsin 3 may he able to block the bacteriai activitme. To test this b3othe~S woumds wvere created, iected w ith £5o rea and treated with s\KA.G as dcscrnbed above, The wounds nwere either treated with a defensin 3 anybody or an isotye control: one 79 appheno0 cacli day for three davs Wound sections were obtamedc and atnied for i am posi iv Oactena M shwI n in gure 1 A. eions deri ed fri woni irated with %ide feom antibody hue re e ran posutie bateria than those related with sl0yne comroi antibodies. aen ection show n was dciedi frno the wound ar ia dly under the scat, Qualti tad1on of C ' in these womds shows that neutraliniont of pdtfsi 3 pir to sNA treatment in ranr infected wounds result in a siiidaet inrease n bacteria., A that were heated w ith Jn It) isotypt controll show an appYniavate 5-ftld reduetion in \viable bacteria (Fijgure 13i. flakcn together these results suSvt dliat sNANG teatnent not only results in the increased kinetics of woind heahng but also pronores an cndowntzious aundueknld repone and sippons the uSE of this nanotiber a novel therapy to enhance wound healmg whie concurrently decreasing wound infttion, 6,23 Crrndusom 0023,8 'he findings presented have demonstrate that a mann datom derived nanotiber, A AG, may be ed US i nov el and etletve method to enhance wound henhi while concurrently decreasing wound inthechon. The data demonstrates that this FDA approved material, whih. h is prseml used for hemnostais stirnua ten thre eapressit of both atytpe and f~ ty pe dtefens in pnpmry endotheha cellA and an up-rgulation of the ypc mn rii keratinoe tes. (0291 Defsins are an essential opponent othe onat imnune syseo These pePtides possess anti duicrodial properties that are active against gramnvposive and negzative bacteria fuig and many ytruses, Defensm arc s 'a 0 (A kiDal cvsteinewrib Otannpeptids found in mraals mnects and plants that are last ed hai de~ fl rnt ies (u. f and t) hased on their pattern of disualide bonding -Jefenin\ r thoughtobpcW to xi to neutrohd are found in vary high _onemrataons (compramn approinnatelv '% of tie total cellular protein ((ian T and R L NO ehrer, IL urr pmn linunol n i0 ' nd are secreted drimy antimicrobial responses (Gan. T 1, 1987, Infect Iimmn 5 36N.71 t has aiso been shown that rabbit allveolar (nacrophages possesscodefen\s s iconarable to rabbit neutrophil (G(ana. T, etal 199. j immunrl 14 H M d I d-defens re fumd in epitheia, cl typesuch as eratmnoevtw mnucosal cpIhdial celLs (Hnade, J, et a, 197, Nature 387(6636:P1, and Far der, J. , t ,'1 , J fbI Chem '% .h) 3) oral cavity nts0ues arid sahm ar- sections (Mathews NL et alp . 1999, Inteet Immun. ~61 )274W~ and kidney where tby can be up related iu esCponsu to m1eious or milamunory stimuli Ganz I and RA, 1ehuer, 9( mr Opin ummunol. N(4:5844l Human fhdefensim 1 i h)FI1 is one of the most iunortnt antrnmerobad peptdes m h e ba tastes De'nsin ex region and secretion could he e~xtrcnh i importantt for cratn w ound therapeu 4 e\ I he antmiembial action bw defense is considered part of mnate immimm and is uon-SPredc and broad spectrum Therefore aepured bacterial resistance as seen wib the oueruse of antNoes. is not an. issue 1O0260 The data presented here also demonstrate that both h a nuro and in v Ait is required for dcfnio expressom sn G eatment decreases Sk: in section ot cutaneous wounds in w ild lype control animals but not in simarly treated Akid I null animis, I is alo important to note that sNAG stimuation of wid typo aneous wounds resuhA i an inetease kinetics of wound closri, Antibody blockade of f defensin results in a reductin in the A\ antihac.i4al data it\. Taken together these indogs sugest a cenital role for Akil in tl regulation of defensin expresion that is responbior the clearance of bacerial infection ad that sNA treatent acti\vates these pathways in wid type anomal 1002611 he data that sunests that NAG treatment of infected nouds could drasticaly decrease bacterial load im patients at least in pant. by 0he induetin ot'deesm pression Sandhv& ccn a rta. is a bacteria frequentl\ found eolotiing the skin and in the nose it is stil a common cause olnosocomid infections often causing postsurgical wownd infeWtons. murer inteetions in hospitals have plagued healthcaxe workers for years and the w idepead usdae of' anthieltcs for treatment has lead to anibiotic resistant strams The dat usented nerem shows that treannnt o Staph infected wounds wAh \ 4G dramatically erased the bactenai load. For cample, the lack of dark purple gram stainmg in the treated W\ mice n Figure's I IA and I I B indicates that the S aureus infxton has ben cleared frm thee wounds oft tle in vilto and w vivo data pvItkie strg e\ dence for the use of TalidiM A inth1e treatment of wounds to decree bacteri infectin and therefore enhance w ud Iedin. 1002621 controll experinnts indicate that the antieno "biso. a direct interaction of th material w ith the bacteria but is Jue to downstreant af'ects such as the relation of defensins by Akt activation it is widely accented ht defensins are imporMt plars in mate immunny and function in antimierubial activItes Most of the evidence for their fiction a the direct i lime of baetwria by W mm ming experimens with pnied defense peptides (Selated, I and A J00uellette. 2005, Natliunol ft j:Mil,) or i smliad iXprmeS iS shOi i Figure II w!nh ireA appheton h o thc pti ed ActA Nc pide The data here snow ha an induction of dcfensin expresmn in wiId type aninis usinir a topical applcatin of sNG results i an annbacterial response i has recent been shown that uangemc mouse models express the human defense gene c restam to Y 9phuii an infection that resuh in death of wild-lype ammals isaleman, NJ A , et al 2003 Naure 422093:1 P5 6) again sugextOng the unportance o deTensins in the regulatton of the ant1)irmrohid rexponxse, 1002631 it has been accepted that the ouhtyp: ostdetesms are spentca v rete m neutrophilsw h. reas the Vty pe defeniins are epithel ial f Origm ftype defensin expression induced in response to xNAG in human kertminocytes both m culture and m tW cM un eous wound hearing model wa detected The is NO data Istrates that idefensin 3 is mainv expressed m the suprahasal ayers after treatment wNh \ i Tis is consisted nth prexus dat whicb locahaed human P-defensoi 2 to the opinous and granular vers of the skin. (Aren A e A., 003 E MI l Pathol, ~4 1M0 "h skin is in constant contact with u vry and mAecton and ftuetions not only as a meehameal barred but also maintains the ability to mount aetve defense agtainsin frtion The exprsm of deensn in the outer layers or skim supports ther role mn eutanous inmate iiimvty However, the data show that sNAG speifically siimulat the expression of three different e defense (1, ed WA n 5 ndothcha cell'. Th' is shown by R'lPC'R, gene arr ay analysis irnrnuuofluorescen. e and L ISAN (d za not show~ a I The iteaction between endothehai ells and leulocytesin usue rypanr N one -f the mui'd and rmost imeant~ tep in uiund heating The procco ~\ fl n as a of Ienkocyte-. from the \vasculature is Liiated by chemotact fat therforefit 4iN inturestingi thait nadefensins are tnidtl by sN AG and may contribute to the necessary weifroph.i. endothelial cellular inmeractns. \oe reemtly. it has come to lintthat slctensius exhilit biuoloveal act iie beyond the inhibition of microbial cellsA including tnen' contibi on to the adaipulle immnune response by eaiitung chemnottc active as' on da'ndrn tc i hert, PR, et iiL 200'. l'ASB I 21(i vTh5- 75) and T cells monoey tes snd macrophages (Garcia, iR., et aL 2001, Cell Tissue R es, 306(2r 257-n) limd keratinocy tes (i ionsaba, hi, et al ,2007. 1 Invest Dcrmatol. I 273 4594604 Previtous eork showsa that human bhta defensing I and 2 havea the ability to cehemnoattract innatnre dendritic cells and T cells through the CC'(~eemokme receptor n (CCR R6) (Yang D. cie aL 1999 Science 286t5439)525 8 and that human beta dcfensin 2 can RT-POR- S :x emoattract NcTNF heated nertrophtl daa the CR6 receptor (Niyonsaba, F . Ogana, and Nagaoka, 2.004, immnunology Jfl " 1 ' , Hua tees 4' an4 eeas enso zo induce chemotaxis by ieractmg w i ( R2. a receptor expressed on macrophages onocvtes. and nenirophis (Rhrl. a, 20of A , i.mno. 2010 ) internyly. the data sow that slAG trament mduces both a' and |Idensmin expresion im endothelid cells. Tken together. the recent data suggest that defensmns max mediate wound beabdog not only fi their properties, but also by bemg chemotacte or oer cell types necesaryior proper heahng. However. applicAtion ot |'defeitns 3 ale did t result mn 3i ate w nound closure (data not show n) anpmg that topwal applicanin of a single defensin toe not sustaim the cluidar ieractions required for increased chemo attraction, cellular recruiment and womnd }Q2641 The. in n data using hoth 'wild type and Ak il knockout animals confirrns the requirement innsi n 3 expression. Since mouse leukocytes do not express er defensins like rnost other rnamnian leukoeve\ Gn Lre T2004. C R Biok, 327039-91in rM ordefensin stains of infiltrating immune cells was not possible. T'reatmnent of airway epthetlal cells n vt w'vth alpha detersins l-3 causes a doe and tnne dependent increased cll migration that requires activtion of P1 K and \IAPK pathw mys, (Aarbiou, . et L 200-4. Am J Respir (Al l Ql )':4-20 1 sNAG stimulaton of endthelial cells has' been shown to musut in the actmtion of vAP1K ('VourJakis J ct aL 2008 2 \Vase Rca 4S3 :22132)und in dat presented acre pharmacological inhibition of MEK also nhibts the expression of the defenssn rm VPTT hese ending suggest that both pathway s impinge on the regulaton of detensin. expression by aNAG;. however, Akt ablation results ina marked reduction of it pN ion both in Virro and in vw. in myeloid eel4, Pdefensmn expression is controlled at the level oVA transrpt. in part, by the E'A(amiy member Pt (ynea. M.. ct atL 2006 J inmunol 1761i i):6906-i7 and \Ma, )E Q Su and P, T enet, 10%. J Biol Chem. 23j15yX7274V ) PU. is a downstream tmget of At :i the B-Cl Ineae. (Rieske, P. and J11. Pongubala. 2001, J iHo Chea. 27t 1 : l60.) in prmrv endothclal CIlls it has been shown that ANt i s upstream of tt both in 'NOV md in vn durown Vos'mphi tracheal development (avnbu KR.et a. 2003. FAS-h 1B 29 N ( rimatioofendohe W ell results a in sed e o tl(probably through At11

A",

which ii regaired fior the nnratinm ot endotheiai cls (\'ournakis. J.N., et al. ,200ii , \'se R es. 45(3)1232E) 1002651 Thus tar, JAG treatment h resulted m a scenes downstream ainlvtties hemosvsts. eell ngann, cell pwhfetion creased wound closure, and as deseibed here, stmuaion of the Mnate imname response resuming m antkbacteri tions 1002661 Given the dramatic increase ofdiabene patients wtin the population who present uwith chroni womids and complications due to wound in'ection, non clinical threatens are in high demand Here, warime derned pGeNec nanoi~bers are descrdhed th ortonmeawe the kinetics of wound heang nut at to stimulate inate inmmity thus providig anttrial activity. The obvious importance of theso observatons is the apph cann tosocoial infections. Of the nosocomial infections surgical wound iecsons predominate ith statistic. samWum; uip to 8% ot'adl surgiceal patients. The direct. cost of rhmse 've pe of jH ficuions is approximately .5 hdbIon dollars per year. Given that defenses are part of the innate mnnune syStCeu aCtivation of these Pathwavs wi il nrechde the generation of resitmt Catanisms as well a allow for the dant't'otvundependein ciaran ce of bacteria: infection WLe of sNAG in nos,'al setting would dera rmch of the nost and m"arkedly rduce the production of amtbiotic resitant species Taken together, these hndm suggest that these marie deriv ed plle\N nanofThIor -us ill be high.\ beeIcial in th~e cinical arena, lO9267l I. Thexample demonstrates that sNAG nanoahers have an antibacterial effect auins [00268 Weia. and lhds: \il ype TOS 6l i7mate i ice hetweemn o 812 weeks Acl were wounded created using a Imm bps punch ( ilteN to rea te two identical wounds on each flank Mice were anesthet/ed Usin an Otodlrane vapor/msn anesthesia machine Vetiinp, tne,\ totlurane w as used at 1% 'r inducnon: 2% for surgery. Prior to Nurgery hair 55(a5 retateiOd by Prc 4oubomne ng c \wre pied and ultured osenight at 37 and adjusted to an absorhtince of Ojt- (0 w Eh w ound was mnculated nwith: i N E () ef'uwoupd of P, rlegnnA f4er ANY30 utinis post noculaton wounds were either treated wx U Gitb membr~ane moiten'd with distilled water test group. nwt or eft untreated (control group, n () On day 3 animals were enthamred and entire wonds w ere harvesed ineludinr the arroundmi ski usig an 8 mm n s punch (lteN One wound per anmd a as VIxed overn allvt nT 4% paraformaldehyd cit 4'' emabeddeti 'al an and sections or analiS, an the othec n found n cultilred and phlaed on LEB media w iluhout tibiout -Ir btoeal guanaion For ui nn wund sections were plcod in G5ml bacterial media an meubated fi 30 mm at 3TC n hile shakmne Colons fo~guvs( w eeuntnated usirs a dilutun senes plated ov ernmghi at 3C. Number of colones per plateper ilti on were conmed and CPU/mi were caulte 1002691 Ro/: The enicacy of sNAG teatment of wunds infected w ithnuam negative' Nacten was assessed, As shown in Figure 14, at 3 a pot iTctin material nunter ts markedly reduced (moie than 2 fbld) in amnds treated wih sN2A mn compunwn to omreated aimilk Tlese tindkgs sggsest that sNAG tretimen results in a mari-d redcttIon in trial load of gram negative bactera, and specifieada / aeuru in infected cutaneous wOunds in addition to reduction m bacterial load of gum positive bacteria shon n E-ample 2). 6A N4oEnf a Number or flins 'Loll RceorGms 1002701 This example demonstrates that a numoer oT derens ims and Totpke recent r reautated by s\AG treatment o hianum endotbelil cells. [002711 Wr/t and kTh i iR Human Chip probes we re prned on epoy sldex HU\EC cells were culured as descrbed in secton 6.2,. and treated w ith sAG nanofiben ("sNAG t for N5 hours N NA was extracted with RN kVol (Telest, Ie. 0 tollowAg inmmihturcr 4 mti rieens adm'pldtred using Amino \llyt MessaeA'kP *1U arena\ 'mplineation kit ( Nipphed Eatems and labl Id The sldes were prepared tor hy brdinnon w W a NA by soak ig n blokmg Soluon Sgm' nris.-bufred sale pH10A in i0NW dH: 0, 1% BS8 \ \B ) ) (bt at RI' 0/N then rinsed and drA ed Samupics containing lbeled target aRNA from sNAG -treated cells were hlyrndied wr ibh shde\ (MO M N , en e nan'd at 955C s 5 rnh bidied mir 48 hours at 34 in 0.1% SDS and 5 N 8SC and i% BSA rclnsed and dryed. The shdes were scmnoed and hybrid ion. det z ina g PerkinJhmer Setn At ray equpmeni and ScanArray Express softw are 1W 3A updated, to identusip iegtaulate d genes. microarra data waus analyzed usin Aglent Genepr'tspn (A\ v 1 . intormion D Analy. 1002721 Genes of mnteest analyed L CFA AM , SPAGI I, defenusins 'DEA O'-i, defense,. and "DEFB< ihdefensin); Tol -like receptors vR4R'. S IGIR R(Simgle tO IL -1 V rated receptor) and TRA F (TNT reeptor associated Faetor 64 Positive econtrolsI 17 (ITyrosine4onohdroger ptry ptophan 5 monohydrogenase action protein) GA PD {iglycraidehvdes phsledndoeae R P1 AA <I Ribotml proti ' I Aat uvLiW (Ihiqwdin C& M'CTB i Aetm B). 1(02731 Rcsd>i: R esils of die nuiroarray gone chip analyses and QPCR validanon of memarni reSu ts are presented in fables UIA' behw sng a custom Mna dup A as determined that a number of derensms and ToI hke receptors are rregulad by NAG treatment of human endothelial cells 1002741 olllike receptors (lLRs) are highiv conserved receptors that recognize specie molecular patterns of acteril component s leading to aon of umate mmity, lworeti ngl y Ormpjb lack an adoeonmune sy stein but are ill resistant to ondrobial infeetOn m mlrcIL. and A Hoffmnann, 200. Curr Opin Werobol, 3(1 16-22,A 'his host defense is th result of an innate immune system that prov ides protection by synthesizing the animierobial peptdel Coll andn 11wheeler AInch are induced . l. Bt et aL 199'6, Ccl (6:973- and Williams A lN, At al. 1997, 151BO 1 1(20:6120%3) Recent w ork has also linked human deenin xpsSion to T1 actiraijon. HLwnan jdefeNsin 2 was shown to be induced in arway epithelial edlk in a TLR-2 dependent mann (Hertz u C t al 2003, J lnnumo 7(12): p 6520-4) TM4 k receptor 4 has bee shon to mediate homam p defbesin 2 inductions in response to (hbmydia pnmtniai inmnofcytes, (Rmano (arratelli, C. et al., 2000, FEMS imunol Med Mierobiot. 57(2) i 4) importady, the P13K/Akt ptwyis a key cormponemnt i I Rt si gnat transduction, controliinga cell ular responses to pathogens. (\\eibhart. . and U T Saemann, 200, Ann Reun Dis 67 SuppI 3:ii-,4,1 Since it s kon that simulatin of TLRs can lad to increased deleim synthesis thisork suggests the potenual fr sNAG as a stimlulator of innate immunity and bacterial clearance va the aciiSation of Ak. Table I list Af s\Ome gen es up-regu lted htN response to SNAG stiuila li ILI_____Prontiammatorva involved mn munedefence CEACAM'- eladhnsion molecule hch direckips' ceayosis of several bactensi aecies SPAG 1 ~ ---------- - li--olcl-t---k isart iro i bcnln A cedes of defensiris thtahrhuth oi e vl,,,, ___ .... 6vs ", .1 ..................o....t.o............o..c...u..r responses twvard infectio ENEUGAND/FUNCT N FOLD I INDUCTION TLITriamA -0-t Iiopptd.o bace I . 86( --- ----- -- ---- ---- -- -- -- --- -- --- -- --- -- ---- t- -- -- --- -- --- -- -- H-- --- -- --- -- --- -- --- -- --- -- --- -- --- -- --- ---- -- --- -- --- -- --- -- --- -- --- -- 'B---- IT -- th ti----------___ _ __ __ __ __ ----- - ---- --- ----- 1 6 ZICR8- 7~ eetrrlte L ouao (HUINE Repvanse to sNAG 1$ugftni Of hours) Gene Name f'ho WI' iPJVECI10s 48b$I nmntz dFfd DEFA41200065 4.2 (43to 194b) --- .S .12 0 0 5 5 4............................. . 5...T (3.664 to 6 123 ) DEEBI YT8200004191 2. 7 ( 1. to7' DEFR 03AJH3000604, 9 (7.4-- to 12 5) DEFS! 110 3000070$ 62 (4 6 th4) DjEPh 23 M130022 397. 791 tk) ) DEES 126 (8200012496 to 2 (&286 to 0 ACTE UMAN(830006231 &8 (5.603 to 7 2A4' RPLI3A~~~~~~ 1psOTCOOl 9. 731 t12 U1 1U C. [80014214 7T2 §5789 to 9 979)' -437 2UMA op~s t 0400003$j0.6 0.4to 0844 -fable 114 DECBOkroari GcValiitation (All Pria 7{Wi1; VAG (Ioiig/nil). lb ECfr 5 Ii) Sample OEMB 1433z DEFB3 h.Cttreaed - Qn DEFB3 1433 ~C unreaed E-Oafve~ to igureaed MCI 1171426 22±1711uMAtreAte.2dA tratd 0$±i0 l7$407 22 4010 18~2z0 (1,20030 [aide V ThWLk ecpos iror Geme Fxpiezssion GeneR [oNme t4~FWCag StGISR ~ ~ ~ ~ ~ . .. ~VOOOO41 .Z9 .3~ .e ... TtM ~*43OO............f t a... T~~~tt4~!. ..... 0406 ... (I2$tog e T&R~ t42000SM~l3 ... a....... .... ~ TtR7 tH 000.... .... .... .... ) TLRS~~~.. ........ IS .~5 .aanz ThAF6~ ~ ...

O.O4S .... .... t .7.

-14.33ZQ.yUMAN 0VOTOOOS 0373 (OA4 to e.8R4) -~ 45 *-~" N CN~ 'N CC "N' Cs ~N N' s"N N".' s" -V v'-' N' N Cs 0 " & N N' "Cr N '11 0 " -~ -C 'N NC' N' -, 'N. Nt N 4 A Cs 44 ~4 C N ~ U 1 - 'N U g SC' 'Cs N N ' " 4 ""C ~ 'r Sc a S.' ~N' 2 V NQ ~t~" ?'N C'N CS' OCC CM '4 a 45 ?- 'C N 'C ~N' ~0 (4 c-NC --N - 'N Ct' (V '4 "C.. N N' S." b-C' 4'N ~ ~ C' SN S C' , -; 'C' 0' 4SC~ "." N '4 u N 'N 4 4) ~ C' NC' '4 .5 SC '4 "N N'; -; ~N N 4) 7 N' N' - ' 5 '4 '4 - N St 5' N' 'N "N (N 90 6A Fxanwle56ANAGand Lnny er NAG &llkrk h TrGae xresion fies 02751 'This examp demo siaies that sNAG nanotibers dier fmm long polkNAe fibers An their dect e expresso, ad specinia in hei eidoeression o ome of the: defensms and "Th like receptors 1002751 iaulab and VThi:i Human Defenmin Chp probes (conentration 20uM quantity 1-2, soh ent SSC hased .p o bu e e prmted on p d .S tCehI es. H IV/C and Ha( eel u ere cultured as described n section ! 2, andi traded wit eniher long fibers C'I \N\G') orQNA( nanotibers ("sNAG> fio 2 hours or 20 hours. RNA vas extracted with RN Vo I Teirsi, Inc) Slow, n manufuer instructions, and amihied usg AnKno Ally) \es! d Al sRNA amphtiealon i '\ mh d Bls)steNns). 0r , RNIA amliiadon, aRNA from cell st td wuh L NAG and aRN Ai frm cells treated with sNAGC wats dttteretially labeked wx oh Cy s or C). r tlrescrtemIC ye r slides w ere prepared 0a hybridiation w ith a R M by soaig in bockmg soiuton (Sigmy Tris-bueired saline p O ,n 1 000m dKI:(, I %BSAW Y. Na, to 005%) ia RT ( N then rnsed and dryd. Samples contammng equal amounts of differential labeled target aRNA from LN,\G and aN WACreated celswere ixed, bybriderrd wh d the shdes (6u1/ide; denatured at MY' fbr 5 mmin; hybridized for 48 hms at h'C in 0.1% SDS amd 5 N SSC and 1" B.AX rised and dryed. The iSlowing exemplary graphs in Table \ I illustrate experimental set up: ... ..... .. . . . . . ... .. .... . . . . . . ...... ..... ) L C .. .r .' .~ ... . *+...... ...... N r* t-4 ' t...........' .... ......... .N ........ ... .........

'

4 ........... . .. ... . .. .... .... ... ...... 54 N> ~ ~ tCN........ .. C' 'a ~ ~........... - ................. ............. N ' .l N tC C 1002771 The 70 des wtare senamed anid hVIbrdion detected usini Perkm-Flmer Sean Anav cg iqpruent and SecaA rran Epress snoa- are \ i.0, updated. For each slide, Ci (13 and cormpntem fluorescence was vsuaied, To idAnt un-regulated and down-eu lated genes nluert'td\ dall sd analed cutn Aeet (vvneSpril GX \.11 Bl ioUfornictOn Data ,\nal a s Genesl interest anayzed DEF A, DFI \B DEFAL DFE4, DEPAN DEFBI, DPfBOlIA D IB104 A DFFIF OSBDEF[IOD, DEP81 11 D11F31 B, D 3 18. OIE 119, DFIZSI PD ltB124, DEFEii 5 PED126 DEEB122 DC 28N DF.-B129, PEEB[i 1. and DEE}34 (DEP \ ' ode'ensm. and 'DEF Ff"bdetenn 'F lL RI, 'FoR 0. 12, T1.kJ, Ti R. f1R5. T R 6, TEl R 7n an TL R C TI R'' Tell receptor); SIIR R (Single IG 11l -relanedt re eptort) RAK2 (l I r ceptorassocruce kmase 1 R'E6 FT N receptor associated factor 6), D 106A (pVdefenm 0)t DIIA (D-fensin 10 ) Negat\ve cotros thee random sequences u 2'3) Posn tC co nI s, 41/ (T yro nme YrnOflty ndOg-Ona$ Ctryono pyophan mnoInoyro gena. aetubon puroten CAIN) (glyceraldexd .smpbosp ite deiw drogenrase 3 RPE LIA (Ribosomal protein LI 1 at 11W0 L0bquitn C), A('TB (AL in I 1002781 Rwi: Results of the mcouany gene chip analyes are p nt 'Tbles \IIl and IX b1lw v Table VII shows ene expression m hnan umilial vi endothehial cel ('MI EC') ater 2h or 24 h exposure to either a)NAG Tbers or ' nanoileXrs '1able L\ shows gene expression mn human keraitine e cell lme (MaCi) a 1der 7h or 24 h exposure to eter [NAG fibers or sN\G nanofibe - I o results denrate that gene expression profile indued bv tone polyNacey giueosumme ic t-es(LN AG' diff'ers from the gcrne cxpresson profile imduced by aN AG nanotibers N %\( '. Specificall) 1> AAG and sN\G differ on ther effet on expression of det'ensm. cenes and I ol receptor genes. nit Q N( - 1 NA(NA-'tj sNNAt4 N33 HUAN0 W4 013 7~132HMN 00h A1 413UMN0140S '1 401 4CR? HUMAN A05 M 1' DF FA 2,S0 3" -1FF\R OA2C OADFA3I 2035 151S186 OS bEFAS- 0.287 ~TAN DER 115 44 MEfII£3 &A 11 DEF9103A 4Th A6 D EFM0I3A 0 628 1348 DE904A 1 2% 2. 61 P)EEOI0A 0 344 9.605 B 0616 DOW1 DUMBIOS UM 3 DEI08 2,Z10j0:41 0616088 U5 1 SO95 Dr1IP4 0000 03367 0616114 A1862 11993 D£f~I 0 14 03331 06F118 1456 U77 ................ ..... .6 .... b...... 3 % 0611242. 14 or? 066619 1M1B 1530 06825IRLA6122$'39 1 0/ DEBI25 1169 ~ 0 F2 0628 05 3

DLF$

1 9 2.26 0DE061129 19 00os IPAEt? 003 1, 11)6 064? 04 .. l3 .. 7 1. 32. .... 3 0. .. ... 0.. 0<3 aw A78 190A R PL, 061 1069 1.82.RM 01 " '1.6 R, 0Y5 1 81 SKA2. 134 A4CO -D ~1 P it2 R I1 ------- I4 ARS 1.333 Q528 TLR6 n -246 -A.482 mg O1 &SO VI9M R B C 0 RF3 09 AS T, TAW ITe I.Jneorm EMM Ata one 1Human ) Lim Fo6d C 0234 0 A OAZ71ACT282 [ 1433Z00 00 + NgaP 0 0041-01 N) 0)00 0.000 PL 13 0~ 1532 0.698 RPL1IA 00900 r-l S87 ... ..... -.. 6 -S --- 3 ------------- \t7IB 13 0A447 [ CS 0330.98 at000 0.000 DEPB 1 26 0348 137 DE 000 04 D iF 9 -0382 4 OEFA2 [0 2 93 6. Examk 6: Fifct of Iradlation ks NG gembranes 09279t I m m'od 'Ppr ntfas AdG \dr ie I. \G nienbrano dA PSed form mkroalM pGY\ t t ers Produteedz iv Previousl described ie >kournakis er '4 C S .Patext 1o. 5$205? and 5M.9 the content of each of whih a incorpotd heiemh3 reence in ts entirtyx Priethntmroatgae n er cultured mn tumque bWorvtot conditumsn usmsh o deined s F. t £olnk'wmg the har'vat ot' rPcoalga fron high-denSt tuhtem fi notedc ih i a \ste\u thecearutEon bi punticaton procin rewultig m batches of pure fibers supended ;41nr for jeons 0iz Fibes . formulated iiQ ptches b concern 95 and owen drymng and wore packtgd and trieed by pomuaruradottnon, Fiber dimensons daierg 20lo 2noim \ 110 ol batches of fers were i itall\ quali\ eonroled usy chmcladpu a etprmeters and each batch met sutret punty onteria pdor to release inal bates were remcd to be substantial free of proteas me ions and other components. The fibers ere then shortened btrdton to produce sNM membranes. Briefl, the starts maternal conaed ot g f pUleN c slurry at a conentation of 1 mg/mt., The concenmA-ion ofth pbleh Ac sirra was confirmed by filtedns $ mL into a 0.2 m fiter. I S 1 o pGlrk~e slonoammg i~ &: it N c was filtered anti lorimanon of a wt kene The w Mae ake as hon nmsferred io a foil pouch, which is a gamma radianon compnble dotainer and sv te2t0d o "Ci Vt gamma. radiaio, theriddiation conditons were tested forthireuecs on poCkNAc compositions asrefl ected iuFigare 15A. j~li~tOI 1 Y t} O/Th(ZW&U }$H fl ^)frNA .t{ahnas. Whil rl'SdilidOn rteduices C A mnolecubr wecght of p~leN Ae, iradition did not disoturb the microstrueture of the fibers ~leiNiAc was irradiated under diferent conditions; as a dry, Iophilised ma ak as a dry m~embiru: as a corientrated slurry M. 70 andight by volume): and as a dilte shnrry (5 nut A suitble molemcr v eigt reduction (to a molecuar w igt of 500,0- 1,0 000 da tons) wa achieved at ao irradiation dos o1000 y ftr dry polymeramd 200 kgy for m eP olymer (Figure 15A) 00281 The chenal And physical structure of the fibers was mintained throughout irradioia veriied by fared (R sperm (Figure 15H9e)eental assay, and scane ag elcron microscopes (E a aaysis.ctroscopic observation of radiated fiers showed derease in the parte lngh (FiguresC1 and I SI. The majoy of thefberarc ess than about 5 po rdength with an average lengih of about 4 un 67 xamk 7; sNAG Nanofibers and Lona Form p-GtcNA Fibers Differ in lTheir Effects on Metabolic Rate ind Serum Denrivation of tImbilical Cord Vein Endoteilat CeAl 1002821 A/arer a' nd 1 s Pooled, mtultiple:-donor human umbilical cord vein endothelial cels C)E(C amnrcx were maintained at 37T wih 5 (O: ml endot belial basal medium 2 RCambres supplemented with EQ grwth medium 2 SingdeQuots as described by Camnbrex prmeeduire ram .strion w as pertored <at D00% contuenev in RP\-1640 supplemented with 01% fetal calf srUm (Gibe BR L- ) W or 24 h followed by stimulation with V.GF I 5 (20 uiml. R&D S estms) or wath highly purited n~leN \ naiotibers or sWAG nanoiners in sterlde nawor (provided b Mir Polymer bnologi ncs No, Danvers Ns., WA \n0th the arnounts m ied n the tigre descripin For 'nllular prolibraionvinbilitty asessment different assau wene used try pan blue excluFoT bi dtet cell county usig a hemiacv tomet andi an MirTT [3(t5din Lt-Jmeth hurol-2yi)dipThomy hetrazolimm brormdedl Iasy procedures described by the manufacturer (Paronega}. |002831 Relh t 14TAN \ 102841 ;KGlt\ i dJ oect rneo a: Mava. As show n : 1 Figure Adid nt result in a higher metabolic rte as measured by MI1 assays. indic ating that this poly meric miateral nd a no auu marked iereases ;n eclhda pronoi mt 00285\ p \vkc V'rCII9C H fsna ( Ai D ath iLAhW n'' Sr 'enum Sarvmao 0 test it pGicNAe fibers ha a dmrcv effect on R% scruratars ed EC ek n ere treated with V'F or with different enneentralons of GleN c ibrs. A shown m igure 17 at 48 h and 72 h after serum starvation, as compared MuIh the total number of ces plated (controit there w2a about fold r :duCtio . the number el rl aller 4o hi or 72h. A t 0 h, tbi decrta5i o cell numberC aa rescued by the addition of VEF or by the addition of prGlcNAc fibers at either 50 or 100 pg'ml At 72 h, ine dcrease in CC number was rescued by the addion of VEGf or ascud by the akbton of pGleNA fibers at (100 pg/mL These results nidicated that ike \'FGIP pGlNAe fiber treatment prevented cell death induced by serum deprivation. 00~286\ R at: Us G' [002871 W \ i inred 11SK; /nurvee haiMln ' A5 r eaured by \N,1' asways sN\G at 50, 100 or 200 hgl resulted in a higher metabobeic of EC than \EGF (Figure 18), 1002 .\ dd .no prt 1:7 T ur cc!! death indw y "asrum deproviont To test i NA(\ fibers had a direct clbet ono R , serunstarvdEiT cell were treated with \E fG or with different oneentratons ofsNAG tigers. As shown n figure 19, at 4 h after srum starvtion, as compared wih the motad number of cells plated ciontrolk thre w as cout 2-foh reducing . the number of OecP', 1 a decrease in cell number was rncued by the addition of \EGP but not by the addition of sNAG fibers at 50, 100 or 200 pg ml These r t minted that not ike VET suAG fiber tr atment did riot prev ciUeJ death induced by serum deprivation, 97 100)2891 CNchskw Tho above resuts deoistrane ilt ANA()' uokiorqg frrp(ocV inc9ease t0empaboli rate of seinImrved LE 6'. as MTd asC an does nt rescue mamoatost osruNo bioledog in a yp la e l c A bsnon tee 1002901 A test article. oTheg sdNAG clduced as prevoenl dribe itiontra S~lVt. asmal ze.The' tes ' aril was OMplie mcrd by MUMin Polymer T'ec01oloies, nc. 642 ioomtihiiitvl esfing - IL-9Q2'* MEM. -Elusion ITt 1 SO) 10.993-5 1002911 iioomnaihliy of the' testm atcewas tested in o sefbols lSa-nm o cels.Noioi~ialreatiit (Gamde 0) waoseed in Lthe 199clsa $Pu exposur . oh test article Th bere elua rsos obtaied Flont the nuSAMvc conrOl. article rimde A and negative contm article IGrade 0) confirmed the suitabihty of the test system aed n the cratera. of the motocol the test article i considered non-toxic and meetn the reqluirement of the Blution "est. Inmernaional Or'azaion for Standardization (ISO) 10993 -5 guidelines, SeeTa N below 98 Table X: ____________REACTImTY GR4DES 6.8.3.1. Maeral andMetod A S C A C A I' S Intramuscular...... Implntaton.est .. IS - -- Wee Imlataio ("ntamscla Imlntto .... 7 : I t ..... 6.8.3 Intramuscular Implantation Test - ISO -4 Week Implantation 6.8.3.1. Materials and Methods [00292] To evaluate the potential of the test article to induce local toxic effects, the Intramuscular Implantation Test - ISO - 4 Week Implantation ("Intramuscular Implantation Test") was used. Briefly, the test article was implanted in the paravertebral muscle tissue of New Zealand White rabbits for a period of 4 weeks. The test article was then evaluated separately using two control articles: positive control Surgicel (Johnson and Johnson, NJ) and negative control High Density Polyethylene (Negative Control Plastic). [00293] Preparation of Test and Control Articles. The test article measured approximately 1 mm to in width and 10 mm in length. The two control articles were prepared. The positive control, Surgicel (C1), measured approximately 1 mm in width by 10 mm in length and was received sterile. Negative Control Plastic (C2), measured approximately 1 mm in width by 10 mm in length and was sterilized by dipping in 70% ethanol. 99 1002941 P-D e cu Each animal was weighed prior to inpatation. On the day of the test, the dorsal sides of the animals were clipped free of fur and loose hair was removed b means of a vacuum Each animal was appropriately anesthetized Prior to implantation, the area was swabbed with a surgical preparation solution. |00295j DttJdinarin Four test article stips nere surgisaly tmpimed into each of the parauertebral muscles of eah rabht, approximately 2 cm horn the idie a parHael to the spinal coltn and a ppromiavn 2 5 em Ham ach other. lhe test arile strips were implanted on one side of theme In a similar ash ion, positie comrol arnele strips (Srge were implanted in the 1ontrakter' muce of each "aimal wo negaive control strips legat em n Qait were implaned adal.tward the ai to he test arilek and to control 'pmtsite on eiher side of the spe (ot of toutrp A totlof alast eight test rtCe W p and bigf o each contl acle stris areged for evaluation, 0021 P v vDu oidum The anials were ma tn ad 1K a perod of a weeks The m ere observed daily for this perio to ensure proper healing of the implmt ites and for clinical S of toxicity Observations ic lded all clinical nmanifatations At the end of he oberrvation prod, the amnmals were weiged Each ammal wa saenficed by an mjectable barbiturate. Sufficient time was alo d to elase for the tssuc to be ct w without bleeding (02971 Gm O naam. BTh paraertebral muses m which the est or conol articles were impl'anted were excised W too trom each animal k'te musce nisue was renioted k carefully slicing arotd the imnpamn Ptes sith a scalpel and hl g out the tVsue he excised implant tissues were examined cgrowlyhu bwit hout using exc essve mn vasa ei pmneedures that might have disrupted the imegrit of tdIk Vi for htto dthoiogical evaluation. The tissues we re placed in properly labeicled cont aina conta mn 1 0% neutal hu fter ed ai ihn 190291 Whabn:Wzay Follow ingiui m forma hin. enab of the implan sites wNas erised fromm the ass of tisse. 10 Onane O >,. patdmteil a larger f The implat containsg the M ea exaimned mraer oscopea. Each sgit-te w as exannned fbt' wuns of iflammaton, eucapsidatron, hemorrhaging. nxcroi and discoloration using the following scale: 2~ Moderute After nlacroco pc oAseroW on rA) I ni Noia Wi$'L"' ein- i1ju. alda SAC oee oUtsmc coinnuingthe ~mfl4A s've wf trose F(o1 og'. w id; thenxh ai c osti 5 - inipginh - 1 tanacr 1.s"",ea x MManphge 0. Neccs 1032up to 0.5 mm. Ver ini aht 2 11 2. tun Mderat 2 ' 0ma Markd [32The In in dimusu hp edition Test was conducted based upon the ibilowing 1 IS) 1093, 194. ihooga I'aluation oF\edhcal Devices - Part 6: T ests tbr ioa F Tlets After implanaon. 2. Of 1 09&2 20. Bieo1al E\valuatior of Medi v \ices -art 12 Sample Preparaton and Referenece RaterN. 3. ASTy P981 K4, 2.004 nndr Practice for Assesment of (Compatbility of Bomatenjals for srgucl Implants w0h. Respect to f ect of' Marials on Msele and one. 4 ASTM F76344. 2004 Standard Pimaedt for Short Term crecmng of plant Materials. 5 JSC1 2005 ener Reqiremn t Conetence of Teaing and Calibration aborator es j10303] The resis of the t uscar Implnt t T reaua a based uon to lon criei. 1. Cal1culated. idi: Fo-r eamclh imlnesia to tal sore is detrmne. heavrage scom of A tew siteS fOr e-~ nima i.,compared to the averair scr tto owolstst ht anmO avera d rence between tet and control stes for A nis is calcunld and the OWNit llvrioitviq. Rating is assumed a1 sbow: 1.5 MikNo Raction > 9 Moderate Reacion 6.0 Marked ResAction '~ A egatve caculaio, is rerte-d a,,e, (1 2. Modiaon of the Rating sal osver re se calculated ece of bioreactivitv Based on the observation of all factomreg. relative size, patern of response. inflammatory vs resoumtionT the patholog obsmerr may revise the Bioreactivity Rating Justfication for the modification to the ratmg pneented m the narrate report (A desrirve 102 narrative report regarding the biocompatibility of the test material is provided by the pathology observer). 6.8.3.2. Results [00304] The results indicated that the test article was non-reactive when implanted for 4 weeks (Bioreactivity Rating of 0.2) when compared to positive control Surgicel; and non reactive (Bioreactivity Rating of 0.0) when compared to negative control High Density Polyethylene (Negative Control Plastic). [00305] Clinical observation. Table XI below shows results of the macroscopic evaluation of the test article and control implant sites indicated no significant signs of inflammation, encapsulation, hemorrhage, necrosis, or discoloration at the 4 week time period. Some test sites and the majority of the positive control, Surgicel, were not seen macroscopically and serial sections were submitted for microscopic evaluation. Table XI: - w -------- __ _ __ _ _ 1_ _ ---------- itesaea*rn4 .. 7A k7 . ----- ---- ---- .. .. .. .. ... .. .. .. ------ - N' '~\~~A~S N'k M ---- Q .A . ........... .... ... 10 [00306] Implantation Site Observations (Microscopic). Table XII below shows results of the microscopic evaluation of the test article implant sites indicated no significant signs of inflammation, fibrosis, hemorrhage, necrosis, or degeneration as compared to each of the control article sites. The Bioreactivity Rating for the 4 week time period (average of three animals) was 0.2, (C1 - Surgicel) and 0.0 (C2 - Negative Control Plastic) indicating no reaction as compared to either of the control implant sites. The pathologist noted there was a moderate polymorphic and histiocytic (macrophages) infiltrate around the in situ test article that was not unexpected given the nature of the test material. Table XII: 10 *w"'o R"a ~ $* -- - -- - -- - -- - ~ ~ -t--------------K ....... .... ... k,'kw 104 Table XII: 4 WevkLti & u &k~ ~ _ $ --- - - ----- 3 77 L7 0:*, le a0 ____ __ a 105 Table XII: 6.8.4---- Inrctaeu Inecio Tes --ISO-10993-10 .. ~~ ..... A-zs' N 6.. ntauanosInekonT - ISO 10931 [00307] USP 0.9% Sodium Chloride for Injection (NaCl) and Cottonseed Oil (CSO) extracts of the test article were evaluated for their potential to produce irritation after intracutaneous injection in New Zealand White rabbits. The test article sites did not show a significantly greater biological reaction than the sites injected with the control article. Based on the criteria of the protocol, the test article is considered a negligible irritant and meets the requirements of the ISO 10993 -10 guidelines. Results are shown below in Table XIII. 106 iracu tancoOs Tst Skin RCctwoO Sctres M~CI £&tract s-- -- -- IX-- ---.-- - - -------------- eOppio Nm 1 0 W e1 nileed T0 j W INW AI an- A O tt * ww - - ' t - --- --- - - -- - - - - - -- - I - - - - - - - -- -- -- -- -- -- - -- ---- N - -- - ------------ B -- --- I e n-s&K-t{1 ;t0 B4-1: &- p d 0C.0 $j -4 - -- -g- --------- .. .... ~4.e-t) c- t f ----- - ------- SO 119 3-t Lveth un 4 00Q I"o-ko~ TA h and a AM <KWM"= of en na rao hedng 10%een e pi:c anhackn an Tel MKedo thel win Amc o fhmmi t h 0 So nal ( lot ane for icyen M al ndCtoeA (SKere t w0 Jd the letateem~sthe euieent of th!SO 099-10 guide- imes Results arev I'l,'JO s , Table XIV Skin Esamhulation Data -'itrce t a "1 J .u{ -2~ x~ -- ( ---- - - AAA Iden "i yi 7. INIcRPORATION flX RR 1 TCE 91 Al l dtRIllS d-ld Nutatll App 'Cuttb cited hU pPuCiflcciT pow is Af :j Ct axe DS" icorpomtd hy erene ach indidua puba On or pa Ap! or ere r y and indviduN\ inK i atod b e- inHO oated 5l refute cC- the C !r: hian been (enbed min wme de1 by way o idhrann nod 5amp e 1o0C r purposes Of emty of undnrstainol it vl be lead&ly a-arn to h- dnary Akll in the ar n mbhgt of the -Li aiv aparn to! ohos of. Fn teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims. [00310] In the claims which follow and in the preceding description of the invention, except where the context requires otherwise due to express language or necessary implication, the word "comprise" or variations such as "comprises" or "comprising" is used in an inclusive sense, i.e. to specify the presence of the stated feature but not to preclude the presence or addition of further features in various embodiments of the invention. [00311] It is to be understood that, if any prior art publication is referred to herein, such reference does not constitute an admission that the publication forms a part of the common general knowledge in the art, in Australia or any other country. 109

Claims (49)

1. A method for treating a bacterial infection in a subject, comprising: topically administering a composition comprising sNAG nanofibers to a subject diagnosed with the bacterial infection or displaying one or more symptoms of the bacterial infection, wherein (i) the sNAG nanofibers have the average length of less than about 15 pim, or (ii) the length of more than 50% of the sNAG nanofibers is no greater than about 15 pim, and wherein the sNAG nanofibers do not have an effect on bacterial growth or survival of Staphylococcus aureus bacterial cultures in vitro.

2. Use of a composition comprising sNAG nanofibers in the manufacture of a medicament for treating a bacterial infection in a subject, said treating comprising topically administering the medicament to a subject diagnosed with the bacterial infection or displaying one or more symptoms of the bacterial infection, wherein (i) the sNAG nanofibers have the average length of less than about 15 pim, or (ii) the length of more than 50% of the sNAG nanofibers is no greater than about 15 rm, and wherein the sNAG nanofibers do not have an effect on bacterial growth or survival of Staphylococcus aureus bacterial cultures in vitro.

3. The method of claim 1 or use of claim 2, wherein the infection is a skin infection, a gastrointestinal infection, a respiratory infection, a urinary tract infection, or a reproductive tract infection.

4. The method or use of any one of claims 1-3, wherein the sNAG nanofibers are in an amount effective to to achieve one or more of the following: (i) reduce the severity of the bacterial infection or one or more symptoms of the bacterial infection, (ii) reduce the duration of the bacterial infection or one or more symptoms of the bacterial infection, and (iii) eradicate the bacterial infection or one or more symptoms of the bacterial infection.

5. A method for treating a disease associated with a bacterial infection or a bacterial imbalance in a subject, comprising: topically administering a composition comprising sNAG nanofibers to a subject diagnosed with the disease or displaying one or more symptoms of the disease, wherein (i) the sNAG nanofibers have the average length of less than about 15 pim, or (ii) the length of more than 50% of the sNAG nanofibers is no greater than about 15 pim, and 110 wherein the sNAG nanofibers do not have an effect on bacterial growth or survival of Staphylococcus aureus bacterial cultures in vitro.

6. Use of a composition comprising sNAG nanofibers in the manufacture of a medicament for treating a disease associated with a bacterial infection or a bacterial imbalance in a subject, said treating comprising topically administering the medicament to a subject diagnosed with the disease or displaying one or more symptoms of the disease, wherein (i) the sNAG nanofibers have the average length of less than about 15 pim, or (ii) the length of more than 50% of the sNAG nanofibers is no greater than about 15 rm, and wherein the sNAG nanofibers do not have an effect on bacterial growth or survival of Staphylococcus aureus bacterial cultures in vitro.

7. The method of claim 5 or use of claim 6, wherein the disease is associated with a bacterial imbalance.8. The method of claim 5 or use of claim 6, wherein the disease is associated with a bacterial infection.9. The method or use of any one of claims 5-8, wherein the disease is a skin disease or a gastrointestinal disease.

10. The method or use of claim 8, wherein the bacterial infection is a nosocomial infection.

11. The method or use of any one of claims 5-10, wherein the sNAG nanofibers are in an amount effective to to achieve one or more of the following: (i) reduce the severity of the disease or one or more symptoms of the disease, (ii) reduce the duration of the disease or one or more symptoms of the disease, and (iii) prevent the progression of the disease or one or more symptoms of the disease. 12. A method for (i) inhibiting the development or onset of a disease associated with a bacterial infection in a subject in need thereof, or (ii) inhibiting the recurrence of a disease associated with a bacterial infection in a subject in need thereof, comprising: topically administering a composition comprising sNAG nanofibers to a site of the bacterial infection or to a site at high risk of the bacterial infection in the subject, wherein (i) the sNAG nanofibers have the average length of less than about 15 rm, or (ii) the length of more than 50% of the sNAG nanofibers is no greater than about 15 rm, and wherein the sNAG nanofibers do not have an effect on bacterial growth or survival of Staphylococcus aureus bacterial cultures in vitro. 111

13. Use of a composition comprising sNAG nanofibers in the manufacture of a medicament for (i) inhibiting the development or onset of a disease associated with a bacterial infection in a subject in need thereof, or (ii) inhibiting the recurrence of a disease associated with a bacterial infection in a subject in need thereof, said inhibiting comprising topically administering the medicament to a site of the bacterial infection or to a site at high risk of the bacterial infection in the subject, wherein (i) the sNAG nanofibers have the average length of less than about 15 pim, or (ii) the length of more than 50% of the sNAG nanofibers is no greater than about 15 pim, and wherein the sNAG nanofibers do not have an effect on bacterial growth or survival of Staphylococcus aureus bacterial cultures in vitro.

14. The method of claim 12 or use of claim 13, wherein the site of the bacterial infection or the site at high risk of the bacterial infection is a wound, a site of a surgical incision, a site of excised tissue, or a site of surgical stitches or sutures.

15. The method or use of any one of claims 8-14, wherein the disease associated with the bacterial infection is bacterimia or septicemia.

16. The method or use of any one of claims 1-6 and 8-13, wherein the bacterial infection is not at the site of a wound, and/or is not associated with or caused by a wound.

17. The method or use of any one of claims 1-16, wherein the composition comprises one or more additional anti-bacterial agents, and/or wherein the composition is administered in conjunction with one or more anti-bacterial agents.

18. The method or use of any one of claims 1-16, wherein the composition does not comprise an additional anti-bacterial agent.

19. A method for treating a bacterially infected wound in a subject, comprising: topically administering a composition comprising sNAG nanofibers to the wound in a subject diagnosed with the bacterial infection or displaying one or more symptoms of the bacterial infection, wherein (i) the sNAG nanofibers have the average length of less than about 15 pim, or (ii) the length of more than 50% of the sNAG nanofibers is no greater than about 15 rm, wherein the sNAG nanofibers do not have an effect on bacterial growth or survival of Staphylococcus aureus bacterial cultures in vitro, and wherein the composition does not comprise an additional anti-bacterial agent.

20. Use of a composition comprising sNAG nanofibers in the manufacture of a medicament for treating a bacterially infected wound in a subject, 112 said treating comprising topically administering the medicament to the wound in a subject diagnosed with the bacterial infection or displaying one or more symptoms of the bacterial infection, wherein (i) the sNAG nanofibers have the average length of less than about 15 pim, or (ii) the length of more than 50% of the sNAG nanofibers is no greater than about 15 rm, wherein the sNAG nanofibers do not have an effect on bacterial growth or survival of Staphylococcus aureus bacterial cultures in vitro, and wherein the medicament does not comprise an additional anti-bacterial agent.

21. The method of claim 19 or use of claim 20, wherein the wound is an open wound.

22. The method or use of claim 21 wherein the open wound is a gunshot wound, a puncture wound, a laceration wound, a cut, an abrasion, a penetration wound or a surgical wound.

23. The method or use of claim 22, wherein the wound is a puncture wound, wherein the puncture wound is caused by a hemodialysis procedure or a catheterization procedure, and wherein the subject has been diagnosed with hemodialysis-related or catheterization-related infection.

24. The method or use of any one of claims 1-4 and 19-23, wherein the sNAG nanofibers are in an amount effective to achieve one or more of the following: (i) reduce the severity of the bacterial infection or one or more symptoms of the bacterial infection, (ii) reduce the duration of the bacterial infection or one or more symptoms of the bacterial infection, and (iii) eradicate the bacterial infection or one or more symptoms of the bacterial infection.

25. The method or use of any one of claims 1-24, wherein the composition is not administered with an immunomodulator, and/or wherein the composition does not comprise an additional therapy which is encapsulated, immobilized or formulated in the sNAG nanofibers.

26. The method or use of any one of claims 1-25, wherein the composition is not administered in conjunction with an anti-bacterial agent.

27. The method or use of any one of claims 1-26, wherein the composition does not comprise an additional active ingredient.

28. The method or use of any one of claims 1-27, wherein the composition is not administered in conjunction with another active ingredient or therapy.

29. The method or use of any one of claims 1-28, wherein the bacterial infection is an infection with a bacteria of at least one of the following species: Bacillus anthracis, 113 Bordetella pertussis, Borrelia burgdorferi, Brucella abortus, Brucella canis, Brucella melitensis, Brucella suis, Campylobacterjejuni, Chlamydia pneumonia, Chlamydia trachomatis, Clamidophila psittaci, Clostridium botulinum, Clostridium difficule, Clostridium perfringens, Clostridium tetani, Corynebacterium diphtheriae, Enterococcus faecalis, Enterococcus faecium, Escherichia coli, Francisella tularensis, Haemophilus influenae, Helicobacter pylori, Legionella pneumphila, Leptospira pneumophila, Leptospira interrogans, Listeria monocytogenes, Mycobacterium leprae, Mycobacterium tuberculosis, Mycoplasma pneumoniae, Neisseria gonorrhoeae, Neisseria meningitides, Pseudomonas aeruginosa, Proteus mirabilis, Rickettsia rickettsii, Salmonella typhi, Salmonella typhimurium, Shigella sonnei, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Streptococcus agalactiae, Streptococcus pneumonia, Streptococcus pyogenes, Treponema pallidum, Vibria cholerae, and Yersinia pestis.

30. The method or use of any one of claims 1-29, wherein the bacteria is resistant to a standard antibiotic therapy.

31. The method or use of any one of claims 1-29, wherein the infection is an MRSA infection, a Pseudomonas infection, or a C. dificule infection.

32. The method or use of any one of claims 1-31, wherein the subject is a human.

33. The method or use of any one of claims 1-32, wherein the composition is formulated as a cream, a gel, an ointment, a membrane, a powder, a spray, or a suppository.

34. The method or use of any one of claims 1-33, wherein more than 50% of the sNAG nanofibers are between about 1 to 12 pm in length, or between about 1 to 8 pm in length.

35. The method or use of any one of claims 1-33, wherein the average length of the sNAG nanofibers is from 4 to 7 ptm.

36. The method or use of any one of claims 1-33, wherein more than 50% of the sNAG nanofibers are no greater than about 10 pm in length.

37. The method or use of any one of claims 1-36, wherein the length of the sNAG nanofibers is determined by scanning electron microscopic (SEM) analysis.

38. The method or use of any one of claims 1-37, wherein the sNAG nanofibers were produced by irradiation of poly-N-acetylglucosamine or a derivative thereof with a dose of irradiation that reduces (i) the average length of the sNAG nanofibers to less than about 15 pm in length, or (ii) the length of more than 50% of the sNAG nanofibers to no greater than about 15 pm in length.

39. The method or use of any one of claims 1-38, wherein the sNAG nanofibers were produced by irradiation of poly-N-acetylglucosamine or a derivative thereof, andwherein (i) 114 the poly-N-acetylglucosamine or a derivative thereof was irradiated in the form of dry fibers, a dry fiber membrane or a dry lyophilized material at the dose of irradiation of 500-2,000 kgy, or (ii) the poly--N-acetylglucosamine or a derivative thereof was irradiated in the form of a suspension, a slurry or a wet cake at the dose of irradiation of 100-500 kgy.

40. The method or use of any one of claims 1-38, wherein the sNAG nanofibers were produced by irradiation of poly-N-acetylglucosamine or a derivative thereof, and wherein (i) the poly-N-acetylglucosamine or a derivative thereof was irradiated in the form of dry fibers, a dry fiber membrane or a dry lyophilized material at the dose of irradiation of 750-1,250 kgy, or (ii) the poly--N-acetylglucosamine or a derivative thereof was irradiated in the form of a suspension, a slurry or a wet cake at the dose of irradiation of 150-250 kgy.

41. The method or use of any one of claims 38-40, wherein the irradiation is gamma irradiation.

42. The method or use of any one of claims 1-41, wherein the sNAG nanofibers were produced from a microalgal poly-N-acetylglucosamine.

43. The method or use of any one of claims 1-42, wherein the sNAG nanofibers comprise N-acetylglucosamine monosaccharides and/or glucosamine monosaccharides, and wherein more than 70% of the monosaccharides of the sNAG nanofibers are N-acetylglucosamine monosachharides.

44. The method or use of any one of claims 1-42, wherein the sNAG nanofibers comprise N-acetylglucosamine monosaccharides and/or glucosamine monosaccharides, and wherein more than 95% of the monosaccharides of the sNAG nanofibers are N-acetylglucosamine monosachharides.

45. The method or use of any one of claims 1-44, wherein the sNAG nanofibers have Grade 0 test score when tested in an elution test, and/or wherein the sNAG nanofibers are non-reactive in an intramuscular implantation test.

46. The method or use of any one of claims 1-45, wherein the sNAG nanofibers do not elicit a detectable foreign body reaction.

47. The method or use of any one of claims 1-46, wherein the sNAG nanofibers increase the metabolic rate of serum-starved human umbilical cord vein endothelial cells in a MTT (3 (4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, and/or do not rescue apoptosis of serum-starved human umbilical cord vein endothelial cells in a trypan blue exclusion test. 115

48. The method or use of any one of claims 1-47, wherein the infrared spectrum ("IR") of the sNAG nanofibers is about the same as or equivalent to that of non-irradiated microalgal poly-N-acetylglucosamine.

49. The method or use of any one of claims 1-48, wherein the sNAG nanofibers have the microstructure of the fibers, and/or wherein the sNAG nanofibers have the chemical and physical structure of the fibers as determined by infrared (IR) spectrum, elemental assay and scanning electron microscopic (SEM) analyses.

50. The method or use of any one of claims 1-49, wherein the infection is a Pseudomonas aeruginosa infection or a Staphylococcus aureus infection.

51. The method or use of any one of claims 1-11 and 19-24, wherein the subject is the subject diagnosed with the bacterial infection.

52. The method or use of any one of claims 1-11 and 19-24, wherein the subject is the subject displaying one or more symptoms of the bacterial infection. 116