!}~_f::JeA~J~t.\JJPI'lii Of' JON LUNOllKR!J AND Eln!~f_}:YflTZllERG
`UNDER37C.F.R.§J.132
`
`\Ve, Jon Lundberg JV!D, PhD. and Eddie \Vc.~itzberg, MD, PhD. declare as follo\:vs.:
`
`L. Jon Lundberg, am a Prot'i:.'.ssor of Pharmacology at the Karohnska Lnstitut{~t (Stockhnhn.,
`l.
`Svveden) and the research grnup leader tbr Pharmacological Nitric Oxide Research at the
`Department of Physiology and Ph:am1acology at the Karolinska Institutet l received my M.D.
`
`from the Karohnska lnstituk, Stockholni, Sweden, in 1993. My interest is in the biology of nitric
`
`oxide (NO) and related nitrogen oxides, ln 1994, me and my colleagues discovered a NO(cid:173)
`
`synthase-independent pathway tbr NO generation in humans. \Vhich results from the redttction nf
`
`inorgunk nitrate and nitrite, l'vfy research is focused on characterizing NO-symhast!-irn:k:pemient
`NO generation from nitrate and nitrite, and clarifying the role of NO in heahh and disease, l'.vf y
`currkulum vitat~ is auw;:hed as Appendix A,
`
`1, Eddie \~/eitiberg, am a Professor of Anesthesiology and lnten.sive Care !\,fodicine at the
`2.
`Departinent of Physiology and Phannacology of the Karnlinska Insti1utet and t±K, research group
`leader for Anesthesiological N'itrk Oxide Research group at the Dq,artment of Physiology and.
`
`Pharmacology at the Karollnska lnstitutet
`
`l am a Senior Consultant fat the Department of
`
`Anes:theskilogy, Surgical Services and Intensive Care t<,,fedicine, Stockholm. During my career, I
`
`have ,vorked on several aspects of nitric oxlde (NO) biology, including cxhakd NO as a marker
`of ainvay inflarnnmtinn. I was a1sn involved in the discovery of the nitrate--nitrite---NO pathway,
`which is n1y current research focu;g. l'V1y curriculum vitae is atwched as Appendix B.
`
`3.
`
`\.Ve are Directi1fS and Shareholders of fkartbeet Ltd. Heartbeet Ltd, has lkensed sorne of
`
`its patents and _patent applications on inorganic nitrates and inorganic nitrites to ThermnUte
`International, LLC ("'T11erm0Lifo").
`
`Human Power of N
`EX1075
`Human Power of N v. Thermolife, PGR2022-00004
`Page 1 of 70
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`DECLARATION OF DR. LUNDBERG AND DR. \VE!TZ:BERG UNDER 37 C.F.R. § L D2
`
`4.
`
`As persons having skill in the relevant field of art and as the main ;,inthors, we have been
`
`engaged by ThermoLifo to provide our opin.ion regarding our puh1katinns on the potential
`hendi.cinl efft.~cts (1f nitrnte. n:m:ndy the foUow"ing articles:
`
`• Larsen et al., "'Effocts of dietary nitrate on blood Pressure in healthv voltmteers," N
`·"
`.
`.,.
`Engl J Afed., 1006, 355(26_}:2791~2793 (hereinatler 'Larsen ::!006"); and
`
`..,.
`
`• Latsen et aL, ''Etlb:.:ts of dietary nitrate on oxygen cost during exerci.'K\" Ada Phrsio/
`
`(Ox__.()., 2007, 191(1):59 .. 66 (hereinafter ''Larsen 2007\
`
`\.Ve understand that our comments should he on the relevant knowledge of the mt up t,:i the time
`
`w·hen the shtdies ,vere published and whl;':ther these studies \.Vould render obvious t(} a person
`
`\.Veil versed in the mt, such as ()Urs.:.+ves, the suitability of nitnW;; supplementation in humans.
`
`The studies of Larsen 2006 and Larsen 2007 only sho,v surprising and novel eftel:ts of
`5.
`ino.rg:a11ic nitrate on blood pressure and aerobic efficiency. !n these articles, ,,.ve do not suggest or
`
`promote to anyone to start ingesting nitrnk in order to achieve better health. The general v:ie,v in
`
`society and srkncc at the time was that ni.tmk: and its generation of nitrite are harmful. by
`
`increasing the risk of cancer and methemoglobt:~nemia in children. This vie,v is still present and
`
`tlK! authorities (including the European Food Safety A.uthority (EFSA) and the \Vorld Health
`
`Organization (WHO)) have not ehanged their recorn1nendations of acceptable daily intake (ADlJ
`
`,md kvds in food and drinking '>Vater.
`
`Resistance by the ethical comrnittees reflected this at tfa~ time, \Ve \Vere only allowed to
`6.
`perfom1 (Hl!' studies using sodium nitrate since the study was fi.)r a shorH.errn and thC'. dose
`
`administered ,vas not higher than \-Vhat couJd be achicv(."t.1 from a nom)al diet (e,g, beet roots or
`
`spinach). V,/e also had to limit the scope of the studies to determining fhe physiological responses
`to inorganic nitrate as opposed to any t1't~atn:i.em t,f d.tS('.HSe. It 'Was hdpfuf that sodium nitrate
`
`·was a food additive in Sv-:eden. and did not faH under the ruies of a pharmaceutical. In other
`
`cmmtries, including the United King(knn and the US, researchers coi:n.numiy chose to use
`
`beetroot juice, a:s a naturni source of anorgnnic nitrate, in order to he able to explore the effects of
`inorganic nitrate. This is likely a major reason why tlw majority of studies on blood pressure and
`exercise are performed \:Vith beetroot j llke,
`
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`DECLARATlON OF DR. LUNDBERG AND OR. \VElTZBERG UNDER 37 CF.R. § l, l 32
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`7,
`
`Thus in order to extrapolate inferences from the Larsen 2006 and Larsen. 2007 sn.adfos,
`one must reme.rnber that nitrates wen.\ and still are, known to cause safoty fa~ue:s .. As not(!d in the
`paragraph 4, nitrate consumption ls stl11 being Hrnited around the w·orld. 1n the United States, the
`
`Food and Drug Administration Hmits sodium. nitrate com::entration to 500 ppm (Exhibit A).
`
`Similar rules limiting nitrate intake are establisl:wd iu the European Union (Exhibit B). Thus all
`
`nutritional advice wai; aimed at minimizing nitrate ima.ke. Some of the justifications for these
`
`limitations is that mcthcmoglobulinernia, gastrointestina.l dismrbances, reduced steroid hnrrnone
`
`production, thyroid hypertrophy, and can::,inogen.k. nitrosamine forrnati(nt fron:1 reaction v>'ith fr(:e
`
`aminoadds \'.Vere \veU known dekkrious effe\'.ts of nltrntes (See e.g,, Vem1eer et al., "Volatile N(cid:173)
`
`nitrosamine fonnation aflcr intake of nitrate at the AIJI level in combination \Vith an mnirn>rich
`
`diet.'' Enriron Health Perspect., 1998, !06(8): 459-463, attached as Exhibit C, and Panesar and
`
`Chan, ·'Decreased Steroid Hom1one Synthesis from inorganic Nitrite and Nitrate: Stmfa~s in
`Vitro and in Vivo,'' Toxicol Appl Phamwrnl, 2000, l 69(3): 222<H\ attached as Exhibit D).
`
`8.
`
`In addition tt) the resistance fro111 ethical <:ommittee, at the time of the studies, there v,:as
`
`;,;ke1,tlclsrn and crit:kisrn from other t~xperts in the fidd/cotkagues, both regarding tht! safr1ty of
`
`nitrate and for suggesting nitrate might have a henefidal effect. Even years after publishing
`Larsen 2006, for exi:m:1pk, there \.Vas the following pub1kation~:
`
`•
`
`\.V.inter et aL, ·'N-nitrosai:nine gent:Tation fwm ingested nitrate via nit1:ic oxide in subjects
`with and without gastroesophagea! rdfox," Gasu-oemerology. 2007 133(1): 164-74,
`
`attached as Exhibit E;
`
`• Panesar, "Dov,r1sides to the nitmte•nitrit,>nitrk oxide path-way in physiology and
`
`therapeutics?"' l'i/at Rev Dmg Di:si.:vv, 2008. (8):710; m:i:lhor reply 710, attached as Exhibit
`
`Land
`• Carpentier er al., '\\fitrate Supplernentation, Exercise, and Kidney Function: Are There
`
`Detdmental Etfoct-;?" Med Sci ,~)>orts .£:..ere. 2015, 47(7): l 519-22, attm.::hed as Exhibit G.
`
`9.
`
`With regard to Larsen 2006, the study did nm make a:ny determinations on the
`
`mechanism of action for ho\v blood pressure \:vas lowered, Neither Larsen 2006 nor Larsen 2{)07
`sh(rwed any effect on ,,,1sodilation. In fa.ct, we explicitly stated in the article: "'Tht.' exact
`
`Page 3 of 6
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`DECLARATION OF DR. LUNDBERG A'ND DR. \VElTZ8ElH., UNDER 37 C.F.R. ~ i .132
`
`mechanism behind the blood-pressure···lovvering effect of 11itraw needs 10 he darified in future
`
`studies." Still today, 10 years later; the rneehanisms behind ihe blood pressure k)\vering effect of
`
`inorganic nitra:te described in Larsen 2006 are not dear, It ·would be erroneous to assume that
`
`just because nitrates .lrnvc a blood pressure lowering e1foct that vasndi!ation is necessarily the
`
`medmnlsm of action that causes the drop in blood pressure.
`
`10.
`
`Din:ct vas,ldilation via NO-mediated ac.:tivation of soluble guanyly! cydas,: in the smo,Ah
`
`musde cells of resistance vessels represents o.nly one possibility. Blood pres~nm,! is determined
`by so many other factors than resistance vessel to.ne, Nitrate and its rnactim1 prnduct:s can directly
`
`affect cardiac function WeHegrino et al.. "Nitrite exerts potent negative inotropy in the isolated
`heart via eN()S-ind1.ipemknt nitric oxide generation and cGl\-:1P--PKG pmh,vay activation,"
`
`Blochim Biophys Acta, 2009, 1787(7): 818-27. ,Jttached as Exhibit H), kidney function (Gao et
`aL -·NADPH oxidase in the nmal mkrovasculatmc is a primary target fix blood pressure(cid:173)
`lo">vering effects by inorganic nitrate ,rnd nitrite," fi.'>penension, 20 l 5, 65( 1 ): 161-70, attached as
`Exhibit I). sympathetic output (unpublished data fr<m1 colleagues in Brazil) from the centraJ
`nervous system and more. Such effect-:; 'INi!l all have a direct impact 011 blood pressure. lt is also
`quite possible that the blood pressure lowering effect was due to the well-knovm nitrate diuretic
`
`action (Se-e Edrmmds, C. W. and Cushny, A. R., Laboratorv Guid1.t __ j_tt_Ji?m.tt!H.!fm.:1l
`Phm:nmf.91~,igy, G. Wahr, 1918 p. 245, attached as Exhihit J; szie tilso Hfatt, "Comparison of the
`Diuretic Effecls of Nitrate Salls \:Vith Other Diuretic,'' Am J PhysioL 1957, l 89: l 73~176,
`
`attached as Exhibit K) .. -'.\.dditiona.Hy. there are previous studies in rats as well as in huma...".ls
`suggesting nitrate to increase blood pressure {siX' Fregly. "Effect of Hypertonic Sodium Nitrate
`Solutions cm Blood Pressuri;.'. of Rats," Proe Soc Exp Biol Med ... l 964, 115; 386·9 L attiched as
`Exhibit L and Porneraz et aJ... "Elevated Sall and Nitrate Levds in Drinkit1g Water Cause an
`Increase of Blood Pressure in Schookhildren;·' A.k.in>J}, Biood Press Res, 2000, 23: 400-403,
`
`attached as Exhibit .tvf),
`
`11.
`
`Additionally, the observed drop in blood pressure in a healthy population al- shmvn in
`
`Larsen 2006 was rather minor. There \Vas no guimi;ntee from Hternture available at the time that
`
`this \vould be replicated on people vvith blood pressme probkm:s. Due to the small mirnber of
`
`patticipants and the homogeneity of the sub\ects (young males), the generalizability of the
`
`Page 4 of 70
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`DECLARAT{ON OF DR. LUNDBERG ANO DR. \VE1TZBf.RG UNDER 37 Cf.R. ii l. 132
`
`findings was fonited, In faz~t there \Vere 110 efft•cts on systolic blood pressure in the study of
`Larsen 2006. Such effoct was reveah:d later using higiK'.f doses and/or larger study populations
`
`(\Vebb t:'.t aL, "Ac:utc BloQd Pressi,lre Lo\vering, Vasoprotective, and i\ntiplatelet Properties c,f
`
`Dielary Nitrate via Bioconversion to Nitrite," lt;pertension, 2008, 51: 784-790, attached as
`
`Exhibit N).
`
`12.
`
`Further studies are still needed to dudd,ite the effect of inorganic nitrate o.n blood
`
`pressure taking in to account age. se~:, underlying disease, etc. These studies need to be
`perfbmied in large cohorts and prderentially \Vith longer duration tJvu \Vha, has been presented
`S{) frir. \\11at is needed to establish nitrate as a robust h!oixi pressure lmvering substance in
`humans is one or preferably several controHed clink .. :ii trials in patients ,vith hypertension. The
`
`studies mtEit have a su.ffident number of patients included, and they must have a nmdn1111zed
`double-blind placebo-controlled design. Moreover, the study dmation slwukl be sufficiently long
`
`to demonstrate sustained effects of nitrate over time. This is important since for exarnpte H1e
`organic r1it1·ates classically develop tolerance over tirne, rneaning that the: t:ffb;;t is diminished.
`
`Even if \,··e were w accept that the hkKid pressure luwedng effect and the oxygen
`13.
`consumption effect could be unequivocally estahllsht!d (\.vhkh \Ve think it could not be
`
`established based on factors such as the small arnount of st1Kl.ies at the tin1e, small a:moum of
`
`subjects in studies fo.g.., Larsen 2007 study only included 9 suhiects). ,:md differentiation in age
`
`gn.)ups\ there were other options i\n m1yone ">vanting theS('. effects ,,-ithout the safot:y concerns of
`
`nitrate. For example, to gain the sarne benefits of lowering blood pressure and (..)xygen
`
`consumption (enhanced athletic performance), people c.an
`
`limit their sodium intake and
`
`supplement \vith ctt:'.atin('..
`
`14.
`
`In short, at the time of Larsen 2006 and 2007 and prior to these publkatkms, ,vt~ nc:ver
`
`rectlmmended, and to the best of om know-ledge nor did anyone else re,'.ommend, that people
`should supplement their diets with nitrate for any purpose. Larsen 2006 or 2007 never said that
`nitrates vvere safe, \,,11ile Larsen. 2006 and 2007 sho\ved two effects of nitrates, the mcdrnn.ism
`by \\•foch th,)se eft'e,;ts occm'!'ed ,vas not knov,m at the tin:1e the stndks \'1,·e:re published. The
`knov,11 correlation berv.:•een nitrates and nitric oxide and the vasQdi1atory effect of nitric oxide b
`
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`DECLARATION OF DR. LUNDBERG AND DR. WEITZBERG UNDER J7 CFJ<. § l. l 32
`
`only one possibi!lty for a mechanism of actinn of nitrate. But we simply did not kno\v whether
`
`nitrate \vould .have a vasodilatory effect ln vivo in lun:nans and :specifically stated the nt.>ed fi.1r
`more sr,udies in Larsen 2006. Therefbre, a person having (}rdinary skill in the art would not
`interpret the results of Larsen 2006 and 20{)7 to conclude that the blood pr1.;.\ssure !<J.vering effoet
`
`of nit.rates is due to vasodilation since other mechanisms could also underlk~ this effoet.
`
`15. We further declare that all statements made herein of our O\Vn kno\vledge are true, and
`
`that all statements made on infotrnation and bdief are believed to be trm.\ and further, that these
`statem,'.nts \vcre made with the knowledge that willful false statemt'.nts and the like so rnade are
`punish.able by fine or imprisonrnent. or both, under Section i 001 of Title 18 of the l.h1ited States
`
`Code, and that such \vi!lfu! false statement~ tnay jeopardize th~ validity of 1.)S patent:; or pa!em
`
`application where their patentability depends on these declarations,
`
`By:
`
`Dated:
`
`By:
`
`Dated:
`
`Page 6 of6
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`APPENDIX A
`APPENDIX A
`
`Page 7 of 70
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`BIOGRAPHICAL SKETCH - CV
`
`POSITION TITLE
`Professor
`
`NAME
`Jon Lundberg
`
`EDUCATION/TRAINING
`
`INSTITUTION AND LOCATION
`
`DEGREE
`(if applicable)
`
`YEAR(s)
`
`FIELD OF STUDY
`
`Karolinska lnstitutet, Stockholm
`Karolinska lnstitutet, Stockholm
`Karolinska lnstitutet, Stockholm
`
`MD
`PhD
`Professor
`
`1987-1993
`1994-1996
`2004
`
`Medicine
`Pharmacology
`Pharmacology
`
`A. Personal Statement
`Jon Lundberg, MD, PhD is a pharmacologist with extensive experience in translational cardiovascular and
`metabolic research. Dr. Lundberg's research activities include the discovery together with Eddie Weitzberg of
`an alternative NO-synthase independent pathway for the generation of NO and other bioactive nitrogen oxides
`in mammals (Gut 1994 ). In this pathway inorganic nitrate and nitrite from dietary or endogenous sources are
`serially reduced to NO in blood and tissues (Nature Reviews Drug Discov 2008, 2015) with profound effects on
`cardiovascular (NEJM 2006) and metabolic (PNAS 2010, Cell Metab 2011) function. Central to this work is the
`characterization of oral symbiotic bacteria as key mediators of nitrate bioactivation (Free Radie Biol Med 2004,
`Nature Chem Biol 2008). Lundberg has extensive experience in gnotobiotic research and the use of germ free
`animals.
`
`In 2010 Dr Lundberg was appointed Distinguished Professor and elected for the Nobel Assembly at
`Karolinska lnstitutet.
`
`B. Positions and Honors
`
`Postdoctoral Fellow, Department of Physiology and Pharmacology, Karolinska lnstitutet (Kl).
`Associate Professor of Pharmacology. Dept. Pharmacology, Kl.
`Co-Founder of Aerocrine AB
`Professor of Pharmacology, Dept. Physiology and Pharmacology, Kl
`Iron Bolt Award, Gordon Research Conference on Oxygen Radicals
`Editor, Br J Pharmacology
`Elected for the Nobel Assembly at Karolinska lnstitutet
`Appointed Distinguished professor at Karolinska lnstitutet
`Chair, Gordon Research Conference: Nitric Oxide.
`Ranked No 3 out of >105 000 researchers in the field of nitric oxide by Expertscape.
`(http://expertscape.com/ex/nitric+oxide)
`
`1996-98
`1998
`1999
`2004-
`2009
`2010-
`2010
`2010
`2013
`2016
`
`May 2011
`
`Page 8 of 70
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`

`

`C. Selected recent publications (5 latest years, 15 selected from >70 peer-reviewed publications)
`
`1. Omar SA, Webb AJ, Lundberg JO, Weitzberg E. Therapeutic effects of inorganic nitrate and nitrite in
`cardiovascular and metabolic diseases. Journal of Internal Medicine. 2016 Apr;279(4):315 36.
`2. Yang T, Peleli M, Zollbrecht C, Giulietti A, Terrando N, Lundberg JO, et al. Inorganic nitrite attenuates
`NADPH oxidase derived superoxide generation in activated macrophages via a nitric oxide dependent
`mechanism. Free Radical Biology and Medicine. 2015 Jun;83:159 66.
`3. Petersson J, Jadert C, Phillipson M, Borniquel S, Lundberg JO, Holm L. Physiological recycling of
`endogenous nitrate by oral bacteria regulates gastric mucus thickness. Free Radical Biology and Medicine.
`2015 Dec;89:241 7.
`4. Lundberg JO, Gladwin MT, Weitzberg E. Strategies to increase nitric oxide signalling in cardiovascular
`disease. Nature Reviews Drug Discovery. 2015 Sep;14(9):623 41.
`5. Gao X, Yang T, Liu M, Peleli M, Zollbrecht C, Weitzberg E, et al. NADPH Oxidase in the Renal
`Microvasculature Is a Primary Target for Blood Pressure Lowering Effects by Inorganic Nitrate and Nitrite.
`Hypertension. 2015 Jan;65(1):161.
`6. Carlstrom M, Liu M, Yang T, Zollbrecht C, Huang L, Peleli M, et al. Cross talk Between Nitrate Nitrite NO
`and NO Synthase Pathways in Control of Vascular NO Homeostasis. Antioxidants & redox signaling. 2015 Aug
`1;23(4):295 306.
`7. Larsen FJ, Schiffer TA, Ekblom B, Mattsson MP, Checa A, Wheelock CE, et al. Dietary nitrate reduces resting
`metabolic rate: a randomized, crossover study in humans. American Journal of Clinical Nutrition. 2014
`Apr;99(4):843 50.15.
`8. Weitzberg E, Lundberg JO. Novel Aspects of Dietary Nitrate and Human Health. In: Cousins RJ, editor.
`Annual Review of Nutrition, Vol 33. Annual Review of Nutrition. 332013. p. 129 59.
`9. Lundberg JO, Weitzberg E. Biology of nitrogen oxides in the gastrointestinal tract. Gut. 2013 Apr;62(4):616
`29.
`10. Kap ii V, Haydar SMA, Pearl V, Lundberg JO, Weitzberg E, Ahluwalia A. Physiological role for nitrate
`reducing oral bacteria in blood pressure control. Free Radical Biology and Medicine. 2013 Feb;55:93 100.
`11. Lundberg JO. Nitrate transport in salivary glands with implications for NO homeostasis. Proceedings of the
`National Academy of Sciences of the United States of America. 2012 Aug 14;109(33):13144 5.
`12. Larsen FJ, Schiffer TA, Weitzberg E, Lundberg JO. Regulation of mitochondrial function and energetics by
`reactive nitrogen oxides. Free Radical Biology and Medicine. 2012 Nov 15;53(10):1919 28.
`13. Hendgen Cotta UB, Luedike P, Totzeck M, Kropp M, Schicho A, Stock P, et al. Dietary Nitrate
`Supplementation Improves Revascularization in Chronic lschemia. Circulation. 2012 Oct 16;126(16):1983 +.
`14. Lundberg JO, Carlstrom M, Larsen FJ, Weitzberg E. Roles of dietary inorganic nitrate in cardiovascular
`health and disease. Cardiovascular Research. 2011 Feb;89(3):525 32.
`15. Larsen FJ, Schiffer TA, Borniquel S, Sahlin K, Ekblom B, Lundberg JO, et al. Dietary Inorganic Nitrate
`Improves Mitochondrial Efficiency in Humans. Cell Metabolism. 2011 Feb 2;13(2):149 59.
`
`D. Entrepreneurship and patents
`
`Named inventor on > 10 granted patent and 8 pending applications. Co-Founder of Aerocrine AB, a biotech
`company that capitalizes on NO as a marker of inflammation. Aerocrine is one of the extremely few Kl-based
`research companies that has made it to the regular Stockholm Exchange Market (June 2007).
`Co-Director of Heartbeet Ltd, a UK based company that owns patents and patent applications related to the
`medical uses of inorganic nitrate and nitrite.
`
`PHS 398/2590 (Rev. 11/07)
`
`Page_
`
`Continuation Format Page
`
`Page 9 of 70
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`

`APPENDIXB
`APPENDIX B
`
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`BIOGRAPHICAL SKETCH
`Prov de the fo ow ng nformat on for the proposed network coord nators and members
`Fo ow th s format for each person.
`
`NAME
`Weitzberg, Eddie
`
`POSITION TITLE
`Professor/Senior Consultant
`Department of Anesthesiology and Intensive Care
`Karolinska lnstitutet, Stockholm, Sweden
`
`EDUCATION/TRAINING (Begin with baccalaureate or other initial professional education and include postdoctoral training)
`
`INSTITUTION AND LOCATION
`
`Medical School, Uppsala University
`
`DEGREE
`(if applicable)
`MD
`
`YEAR(s)
`
`FIELD OF STUDY
`
`1976-1982 Medicine
`
`Karolinska University Hospital
`Resident Anesthesiology and Intensive Care
`Karolinska lnstitutet, Stockholm
`
`Specialist
`
`1987-1993 Medicine
`
`Ph.D.
`
`1989-1993 Medicine
`
`A. Positions and Honors (chronological order)
`1987-1993
`Residency, Anesthesiology and Intensive Care, Karolinska Hospital, Stockholm
`Associate Professor in Anaesthesiology and Intensive Care
`1996-2003
`1997
`International expert committee-American Thoracic Society
`1997
`Founder of Aerocrine AB (Medical diagnostics)
`1997-1998
`CEO Aerocrine AB
`1997-2005
`Member of the Board Aerocrine AB
`1999-present
`Senior Consultant in Anaesthesiology and Intensive Care
`2001-present
`Head of Reasearch Education (PhD programme)
`2004-present
`Professor in Anaesthesiology and Intensive Care
`2005
`International expert committee-American Thoracic Society
`Member of the Board of Stockholm School of Entrepreneurship (SSES)
`2009-present
`Board member Swedish Society of Anesthesiology and Intensive care, Research and
`2012-present
`Education
`
`B: Publications:
`
`H-index: 54, total 12787, citations 2014; 1310
`
`10 highest-impact publications:
`
`1. Weitzberg E, Lundberg JM, Rudehill A. Elevated plasma levels of endothelin in patients with sepsis
`syndrome. Gire Shock. 1991 Apr;33(4):222-7.(Cited 175)
`2. Alving K, Weitzberg E, Lundberg JM. Increased amount of nitric oxide in exhaled air of asthmatics.
`EurRespirJ. 1993 Oct;6(9):1368-70. (Cited 1208)
`3. Lundberg JO, Farkas-Szallasi T, Weitzberg E, Rinder J, Lidholm J, Anggaard A, Hokfelt T, Lundberg
`JM, Alving K. High nitric oxide production in human paranasal sinuses. Nat Med. 1995 Apr;1(4):370-3
`(Cited 550)
`4. Lundberg JO, Weitzberg E, Lundberg JM, Alving K. lntragastric nitric oxide production in humans:
`measurements in expelled air. Gut. 1994 Nov;35(11 ):1543-6 (Cited 426)
`5. Larsen FJ, Ekblom B, Sahlin K, Lundberg JO, Weitzberg E. Effects of dietary nitrate on blood pressure
`in healthy volunteers. N Engl J Med. 2006 Dec 28;355(26):2792-3 (Cited 286)
`6. Lundberg JO, Weitzberg E, Gladwin MT.The nitrate-nitrite-nitric oxide pathway in physiology and
`therapeutics. Nature Rev Drug Discov. 2008 Feb;7(2): 156-167 (Cited 916)
`
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`

`7. Jansson EA, Huang L, Malkey R, Govoni M, Nihlen C, Olsson A, Stensdotter M, Petersson J, Holm L,
`Weitzberg E, Lundberg JO. A mammalian functional nitrate reductase that regulates nitrite and nitric
`oxide homeostasis. Nat Chem Biol. 2008 Jul;4(7):411-7.(Cited 169)
`8. Larsen FJ, Schiffer TA, Borniquel S, Sahlin K, Ekblom B, Lundberg JO, Weitzberg E. Dietary inorganic
`nitrate improves mitochondrial efficiency in humans. Ce// Metab. 2011 Feb 2;13(2):149-59. (Cited 255)
`9. Carlstrom M, Larsen FJ, Nystrom T, Hezel M, Borniquel S, Weitzberg E, Lundberg JO. Dietary
`inorganic nitrate reverses features of metabolic syndrome in endothelial nitric oxide synthase-deficient
`mice. Proc Natl Acad Sci US A. 2010 Oct 12;107(41 ):17716-20. (Cited 130)
`10. Lundberg JO, Gladwin MT, Weitzberg E. Strategies to increase nitric oxide signalling in cardiovascular
`disease. Nat Rev Drug Discov. 2015 Sep;14(9):623-641. (Cited-)
`
`Publications related to the proposed network program:
`See ref 4-10.
`
`C: Current and anticipated grant support
`Swedish Research Council (Nitrogen oxides in critical illness)
`Swedish Heart and Lung Foundation (The Nitrate-Nitrite-Nitric Oxide Pathway:
`Role in Cardiovascular Physiology, Therapeutics and Nutrition)
`Jochnik Foundation
`(Diet, Nitrate and Blood Pressure)
`Stockholm City Council
`(Nitrogen oxides in anesthesiology and intensive care)
`Karolinska lnstitutet
`
`2014-2016
`
`2014-2016
`2015-2017
`2014-2015
`Yearly
`
`D: Previous experience in collaborative research
`
`Dr Weitzberg's research has a strong translational focus and includes studies on the roles of
`endothelin and nitric oxide in various inflammatory conditions; endotoxin shock, urinary tract
`infections, pulmonary and systemic hypertension. In relation to his clinical position at the Neuro-lCU
`he is also studying different aspects of brain injury. His current focus is to explore the biochemical
`relationships between nitrate-nitrite-NO and how this impacts on normal physiology and disease
`states. This represents an important alternative source of NO especially during hypoxia when the
`classical L-arginine/NO synthase pathway may be dysfunctional. An expanding number of studies
`suggest a therapeutic potential for nitrate and nitrite in diseases such as myocardial infarction, stroke,
`subarachnoid hemorrhage, systemic and pulmonary hypertension and gastric ulceration. Special
`attention is given to the role of this pathway in the critical care setting. Also, he and Dr. Lundberg
`made seminal discoveries related to the detection and quantification of NO in exhaled gas. This is
`now a broadly-accepted measure of pulmonary inflammation in conditions such as asthma and
`ARDS.
`
`Dr. Weitzberg maintains an active ongoing collaboration with a key colleague at the Karolinska
`lnstitutet, Jon Lundberg, MD, PhD. Together, they invented the first device for the clinical detection of
`exhaled NO that is now in widespread clinical use. He has been a partner in the recently completed
`EU consortium investigating the biochemical and physiological interactions between eicosanoids and
`nitric oxide (Eicosanox). At present he is also a partner in EU collaborative project Gasotransmitters
`(EU Network on Gaostransmitters). He also has ongoing collaborations with the present co-applicants
`(Freeman, Gladwin, lschiropoulos). Dr. Weitzberg has co-authored several publications with the
`proposed network members Lundberg, Gladwin and Freeman and they have together organized
`several international meetings on nitrate, nitrite and reactive nitrogen species (see meeting report
`below).
`Nitrate and nitrite in biology, nutrition and therapeutics. Lundberg JO, Gladwin MT, Ahluwalia A,
`Benjamin N, Bryan NS, Butler A, Cabrales P, Fago A, Feelisch M, Ford PC, Freeman BA, Frenneaux
`M, Friedman J, Kelm M, Kevil CG, Kim-Shapiro DB, Kozlov AV, Lancaster JR Jr, Lefer DJ, McColl K,
`Mccurry K, Patel RP, Petersson J, Rassaf T, Reutov VP, Richter-Addo GB, Schechter A, Shiva S,
`Tsuchiya K, van Faassen EE, Webb AJ, Zuckerbraun BS, Zweier JL, Weitzberg E. Nat Chem Biol.
`2009 Dec;5(12):865-9.
`
`Dr Weitzberg is not an applicant, member or coordinator of any other Leducq Foundation-supported
`activity.
`PHS 398/2590 (Rev. 11/07)
`
`Page_
`
`Continuation Format Page
`
`Page 12 of 70
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`

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`FDA Home·3 Medical Devices4 Databases5
`
`The information on this page is current as of April 1 2015.
`For the most up-to-date version of CFR Title 21, go to the Electronic Code of Federal Regulations
`(eCFR). 6
`
`Exhibit A
`
`New Searct1
`[Code of Federal Regulations]
`[Title 21, Volume 3]
`[Revised as of April 1, 2015]
`[~l~~: 21CFR172.170]
`
`Help' I More About 2·1 CFR 8
`
`TITLE 21--FOOD }\.ND DRUGS
`CHAPTER I----FCCD AND DRUG ADMINISTRA.TION
`DEPA:RTMENT CF HEALTH AND HUMAN SE:RVICES
`SUBCEAY:.'ER B--FOOD FOR
`l-JUMAN CON~oUMPTICJN
`f.,A.RT 172 ----
`.FOOD P,.DDITIVES PEIZ:.\/IITTED FCil? DIRECT .? .. DDITION TO YOCD FOP.
`
`(CONTINUED)
`
`Subpart 8--Food Preservatives
`
`Sec. 172.170 Sodium nitrate.
`
`The food additive sodium nitrate may be safely used in or on specified
`foods in accordance with the following prescribed conditions:
`
`(al It is used or intended for use as follows:
`
`a preservative and color fixative, with or without sodium
`( l)
`ni t:r:L te, :i.n smoked, cu:ced ::,ableJ::Lsh, smo.ked, cured sa.l.mc:.n, and smoked,
`cured shad, so that: the level cf sodium nitrat:e does not exceed 500
`parts per million and the level of sodium nitrite does not exceed 200
`parts per million in the fini.shed product.
`
`(2) As a preservative and color fixative, with or without sodium
`nit:r:Lte,
`:i.n meat:--·cur:l1v::; preparat::Lons tor t:he home cur:i.ng oJ: meat and
`meat: products (i_nclud:Lng poult:ry and w:l.1d gt,me), w:Lt.h d:i.rect:i_ct1s for
`use which limit the amount of sodium nitrate to not more than 500 parts
`per million in the finished meat product and the amount of sodium
`nitrite t:o not more than 200 parts per million in the finished meat
`product.
`
`(b) To assure safe use of the additive, in addition to the ct:her
`information required by the Act:
`
`(1) The label of the additive or ot a mixtu:ce containit1g the additive
`shall bear:
`
`(ii The name of the additive.
`
`(ii) A st:at:ement. of the cot1centration cf the additive it1 any mixture.
`
`(2) If in a retail package intended for househcld use, the label and
`labeling of the additive, or of a mixture containing the additive,
`shall bear adequate directions for use to provide a final food product
`that complies with the lindtations prescribed in paragraph (a) of this
`
`Page 13 of 70
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`

`

`sect.i.on ~
`
`(3) If in a retail package intended tor household use, the label of the
`additive or oE a mixture containing the additive, shall bear the
`staterr:.ent Hl<eep out of the reach of children H
`~
`
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`
`3. http://www.fda.gov/default.htm
`
`4, http://www.fda.gov/MedicalDevices/default.htm
`
`5. http://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/Databases/default.htrn
`
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`SID=3ee286332416f26a91d9e6d786a604ab&,mc::::true&,tpl::::/ecfrbrov,1se/Title21/21tab_02.tpl
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`Page 15 of 70
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`Page 15 of 70
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`Journal of the Science of Food and Agriculture
`
`J Sci FoodAgric 86:10-17 (2006)
`DOI: 10.1002/jsfa.2351
`
`Review
`Nitrate in vegetables: toxicity, content, intake
`and EC regulation
`Pietro Santamaria*
`Dipartimento di Scienze de/le Produzioni Vegetali, University of Bari, Via Amendola 165/A, 70126 Bari, Italy
`
`Exhibit B
`
`Abstract: Nitrate content is an important quality characteristic of vegetables. Vegetable nitrate content is
`of interest to governments and regulators owing to the possible implications for health and to check that
`controls on the content are effective. Nitrate itself is relatively non-toxic but its metabolites may produce
`a number of health effec