BOTH OF OUR SPEAKERS TODAY
ARE FROM FREDERICK, AND OUR
FIRST SPEAKER IS
DAN McVICAR.
HE GOT HIS PHD FROM THE MEDICAL
COLLEGE OF VIRGINIA, VIRGINIA
COMMONWEALTH UNIVERSITY.
HE'S BEEN AT NCI A LONG TIME,
190, POST DOCTORAL FELLOW AND HE
SUBSEQUENTLY ACCEPTED A POSITION
AT THE LABORATORY OF IMMUNOLOGY
AND HE'S DEPUTY DIRECT OF THE
CANCER INFLAMMATION PROGRAM.
HIS TITLE, THE TWO FACES OF
REDOX AND IMMUNOLOGY.
>> THANK YOU.
WELL, THANK YOU.
WHAT I'M GOING TO TRY TO DO THIS
AFTERNOON IS TELL YOU A LITTLE
BIT ABOUT MY EXPERIMENTS OR MY
EXPERIENCE WITH REDOX BIOLOGY.
I'M RELATIVELY NEW TO BIOLOGY
AND I'M NOT A CHEMIST SO IT WILL
DIFFER DRAMATICALLY FROM WHAT
YOU'LL HEAR FROM THE SPEAKER AT
5:00.
WHAT I'M GOING TO DO IS TELL YOU
ABOUT SOME OF THE INTERFACES
BETWEEN REDOX BIOLOGY AND
IMMUNOLOGY.
THE TITLE ON THE ORIGINAL
PROGRAM SAID IMMUNOLOGY WHICH IS
A LITTLE BIT MORE THAN A ONE
HOUR TALK, SO WE CAN'T COVER ALL
OF IMMUNOLOGY.
SO WHAT I'M GOING TO TRY TO DO
IS SHOW YOU A LITTLE BIT OF
IMMUNOLOGY AND SHOW YOU SOME
REDOX CHEMISTRY AND BIOLOGY AND
HOW IT FITS IN AS WE SEE IT.
I'M GOING TO DO THAT BY MIXING
IT IN A LITTLE BIT WITH SOME OF
OUR ONGOING WORK.
SO WE'LL SHOW YOU SOME OF THE
MOST UP TO DATE STUFF THAT WE
CAN THINK OF, AT LEAST THAT I
THINK OF.
SO, AS WAS MENTIONED, I'M A
MEMBER OF THE CANCER AND
INFLAMMATION PROGRAM.
WE'RE OBLIGATED BY LAW TO SHOW A
SLIDE LIKE THIS WHICH IS THE
CHARACTERISTICS THAT LEAD TO
MALIGNANCY, AND THIS SORT OF
MODEL HAS BEEN UPDATED OF COURSE
OVER THE YEARS, AND THOSE OF US
THAT ARE INTERESTED IN TUMOR
INFLAMMATION ARE PLEASED NOW TO
KNOW THAT THIS HAS BEEN ADDED,
TUMOR PROMOTING INFLAMMATION HAS
BEEN ADDED.
IN OUR PROGRAM WE'RE INTERESTED
IN THE REGULATION OF
INFLAMMATION AND HOW THAT
REGULATION OF INFLAMMATION IS
PROMOTE OR EXACERBATE DISEASE
AND HOW IT CAN BE PLAYED MEDIA
OR TAKEN ADVANTAGE OF IN A
THERAPEUTIC SETTING FOR DISEASE.
AND IT'S KIND OF CONVENIENT FOR
ME THAT RIGHT ACROSS THE DRAWING
HERE IS DEREGULATION OF CELLULAR
ENERGETICS BECAUSE WE'RE GOING
TO TALK A GOOD BIT ABOUT
METABOLISM TODAY AND REDOX
BIOLOGY AND HOW IT RELATES SO
IT'S KIND OF NICE THAT THEY ARE
RIGHT NEXT TO ONE ANOTHER.
SO AGAIN BY JUST WAY OF GIVING
YOU AN IDEA OF WHERE I'M COMING
FROM AND WHERE MY THINKING COMES
FROM, THIS IS KIND OF AN OVERLY
SIMPLISTIC I WOULD ARGUE MODEL
OF WE THINK OF CANCER
DEVELOPMENT AND PROGRESSION.
YOU'LL HAVE A LIGHTNING BOLT
HITTING A CELL AND THEN THERE'S
AN INFLAMMATORY RESPONSE GEM AS
THE IMMUNE SYSTEM TRIES TO
ELIMINATE THAT TARGET.
AND THIS IS KNOW AS
IMMUNEOEDITTING.
THE IDEA HERE IS THE IMMUNE
SYSTEM MAY RECOGNIZE THE CELL
BECAUSE IT'S DAMAGED AND TRY TO
REMOVE IT.
BUT AT THE SAME TIME THERE'S
OFTEN INFLAMMATION ASSOCIATED
WITH THIS EVENT.
IF YOU THINK OF -- FOR THOSE OF
YOU WHAT ARE INVOLVED IN THE
LABORATORIES AS YOU KNOW AND YOU
THINK OF THE CON OSAMA ONTICAL,
AND YOU ADD A CARCINOGEN AND AN
IRRITATE AND IT IRRITATES THE
SKIN AND DRIVES A LOT OF
INFLAMMATION AND THIS IS
NECESSARY FOR THE PROGRESSION OF
DISEASE.
SO I'VE DRAWN THAT IN A LITTLE
BIT.
SOME OF THESE CELLS WILL ESCAPE
AS THE HYPE -- HYPOTHESIS.
THEY WILL DEVELOP INTO LARGER
DISEASE.
SO IN MY LABORATORY AND IN THE
CANCER AND INFLAMMATION PROGRAM,
WE'RE VERY INTERESTED IN HOW THE
IMMUNE CELLS ARE INTERACTING IN
THIS MASS AND HOW THEY ARE
PROPAGATING THE GROWTH OF THIS
TUMOR AND THE ABILITY OF THE
TUMOR TO ESCAPE ULTIMATELY INTO
METASTHESIS.
WE'LL TALK A LITTLE LITTLE BIT
ABOUT A TRADITIONAL VIEW OF
REDOX AND IMMUNOLOGY.
I'LL SHOW YOU WHAT THAT MEANS.
IT'S GENERALLY BASED ON
BACTERIAL ACTIVITY.
AND I'LL SHOW YOU THERE'S A LOT
MORE THAN REDOX IMMUNOLOGY AND
THEN IT'S GOING TO BE CENTERED
ON THE ABILITY OF REDOX TO
REGULATE METABOLIC PROCESSES IN
THE CELLS.
CAN YOU TELL I'M AN OLD GUY.
I HAVEN'T LEARNED BIOCHEMESTRY
IN A LONG TIME.
YOU GUYS HAVE PROBABLY LEARNED
MUCH MORE RECENTLY BUT JUST IN
CASE WE'RE GOING TO OVERVIEW A
LITTLE BIT.
AND THEN WE'LL TALK ABOUT THE
MODEL SYSTEM THAT WE USE WHICH
ARE MACROFASIA SCATTERED SHOWERS
AND HOW IT'S REGULATED BY REDOX
BIOLOGY.
THIS KIND OF INVARY ITSABLY ENDS
UP WITH NOS.
SO SPOILER ALERT, NOS IS
INVOLVED IN THIS.
WE'LL TALK ABOUT SOME OF THE
CHEMISTRY.
HOPEFULLY I'LL BE ABLE TO TIE
THIS BACK TO THE THINGS DAVID
TOLD YOU ABOUT A LITTLE BIT AS
WE GO IN AND THEN WE'LL HAVE A
QUICK REVIEW AT THE END AND
HOPEFULLY WE'LL GET OUT ON TIME.
OKAY.
IMMUNOLOGY IN ONE SLIDE OR LESS
IS RIGHT HERE.
THOSE OF YOU WHO HAD SOME
IMMUNOLOGY WILL RECOGNIZE THIS
IS PAINFULLY OVER SIMPLISTIC.
THIS IS LOOKING FROM A 30,000
FOOT VIEW.
JUST TO MAKE SURE WE'RE ALL ON
THE SAME PAGE.
SITES OF INJURY ARE GOING TO
SENSE A PROBLEM.
THEY ARE GOING TO SENSE A
PATHOGEN, SENSE A DANGER AND
THIS RESOLVES IN MACROPHAGES AND
THESE ARE GOING TO COORDINATE
THE INFLAMMATORY RESPONSE.
THEY ARE NOT ONLY GOING TO THE
ATE INFLAMMATORY RESPONSE BUT
THEY ARE ALSO IMPORTANT IN
MAKING SURE THE SKWAOEPT
ADAPTIVE RESPONSE GOES AS
PLANNED.
FOR EXAMPLE THEY WILL PRODUCE
MULTIPLE SITEO KINE.
IT HELPS TO THE DEVELOPMENT OF
T-CELLS INTO A T-HELPER ONE OR
TH1 STATE AND THIS IS DRIVEN
VERY STRONGLY BY IL-12.
IT'S THESE THAT PRODUCE A KEY
SITOKINE.
INTERFERON GAMMA AND YOU CAN SEE
THAT THIS AUTHOR HAS WRITTEN IN
VERY NICELY REACTIVE OXYGEN
INTERMEDIATES SO IT'S GOING TO
PROMOTE THE REDUCTION OF REDOX
BIOLOGY IN THESE CELLS WHICH IS
GOING TO BE INVOLVED IN THE
ELIMINATION OF THE BUG OUT OF
THESE CELLS AND ULTIMATELY THOSE
ANTOJENS AND THEY ARE KILL
NEWLY-INFECTED CELLS AND THE
SYSTEM AMPLIFIES.
SO DOES THAT MAKE SENSE AS THE
ONE SLIDE FOR IMMUNOLOGY?
AND I THINK I TOUCHED ON EACH OF
THE KEY THINGS I WANTED TO
MENTION THERE.
I WOULD FOLLOW-UP FROM AGAIN A
VERY NAIVE SENSE, IF YOU SAY
WHAT DO YOU KNOW ABOUT REDOX AND
IMMUNOLOGY, YOU'LL SAY I THINK
THAT'S HOW MACROPHAGES KILL
STUFF.
MACROPHAGE STANDS FOR BIG EATER.
THAT'S LITERALLY WHAT IT MEANS.
IT'S A BIG CELL AND IT EATS BIG
STUFF AND KILLS IT.
WHEN IT ZEROS IN IS WHEN YOU SEE
MOST OF THE REDOX BIOLOGY.
FOR EXAMPLE, THIS IS A DRAWING
HERE OF A PHAGOSOME OFF THE
INTERNET.
YOU CAN GET THE ORIGINALS.
WE'LL GIVE YOU SOME INFORMATION.
HERE'S THIS BIG OAT PHALLOW--
PHAGOSITE.
THESE ARE PUMPS.
THEY ARE GOING TO PUMP PRO TONS
INTO THE SRAFPLT -- VACUOLAR.
LOW PH IS ANTI-MICROBIAL BUT IT
ALSO FACILITATES THE CHEMISTRY
OF THE OTHER REDOX PATHWAYS
GOING ON IN THE PHAGOSOMES.
ONE OF THE KEY ONES IS THE
NADPH OXYDASE.
IT'S BUILT THERE.
AND IT'S BUILT THERE
INTENTIONALLY BECAUSE IT IS
GOING TO SHOOT REACTIVE OXYGEN
SPECIES INTO THE LUMEN.
THE WHOLE IDEA IS TO PUT THE
BUGS INTO A ROOM AND DUMP BAD
STUFF ON TOP OF THEM.
THAT'S WHAT IT'S GOING TO DO.
IT'S ASSEMBLED FROM MULTIPLE
COMPONENTS AND THE STIMULATION
IS WHAT CAUSES THEIR ASSEMBLY
INTO THERE MEMBRANE COMPLEX.
AS I MENTION HERE, IT'S GOING TO
DIRECT INTO THE PHAGOSOME.
AN IMMEDIATE CONNECTION TO
METABOLISM RIGHT AWAY.
THE NADPH HAS TO COME SOMEWHERE
MUTATIONS IN THE NADPH OX
NADPH OXIDACE WHEREBY PATIENTS
CANNOT KILL BACTERIA IN THE
PHAGOSOME AND THEY GET CHRONIC
GRANULOMAS.
WHAT HAPPENS HERE, A LITTLE BIT
OF THE CHEMISTRY.
THIS IS AN IRON BASED ENZYME.
IT PRODUCES SUPEROXIDE.
IT HAS A COUPLE OF ROOTS IT CAN
GO THROUGH IN ORDER TO BE TURNED
INTO THE GOOD STUFF THAT'S GOING
TO KILL THIS BUG.
ONE OF THE FIRST ONES IS THAT IN
THIS LOW PH, THE SUPER OXIDE
WILL BE CONVERTED INTO HYDROGEN
PEROXIDE.
HYDROGEN PEROXIDE IN TURN IS
REACTED BY THE MILO, WHICH WE'LL
TALK ABOUT.
HYDROGEN PEROXIDE BY REACTING
WITH IRON IN THE PHAGOSOME
PRODUCTIONS HIGHDROXAL WHICH ARE
VERY GOOD.
NPO WILL BUILD THE HYPER
CHLORIDE.
LITERALLY BUILDING BLEACH.
SO IT'S GOING TO BUILD BLEACH.
WE ALL KNOW THAT BLEACH IS GOOD
AT KILLING BACTERIA.
SO THAT'S WHAT'S GOING ON WITH
THE NADPH.
YOU CAN GET A LOT OF REACTIVE
OXYGEN SPECIES OUT OF THE
NADPH OXDATES BUT YOU CAN ALSO
GET IT OUT OF THE MITOCHONDRIA.
THE MITOCHONDRIA SHARE A
TREMENDOUS SOURCE OF SPECIES.
THIS IS A HUGE REDOX SYSTEM AS
ELECTRONS ARE TRANSPORTED ACROSS
THE CHAIN AND THEY FORCE
HYDROGEN ATOMS INTO THE SPACE
AND THOSE HYDROGENS RETURNING
ARE GOING TO DRIVE THE
STP CYNTACE.
COMPLEX THREE IS A BIG LEAKER OF
LESSON ELECTRONICS.
AGAIN, THESE WILL BE CONVERTED
BY SOD IN THE MIGHT MITOCHONDRIA
AND
SOME OF THE SAME CHEMISTRY I'VE
SHOWED YOU.
THE OTHER THING I WANTED TO
POINT OUT HERE IS THAT A SECOND
DIRECT CONNECTION.
OF COURSE THE ELECTRON TRANSPORT
CHAIN IS CONNECTED TO
METABOLISM.
I THINK WE ALL KNOW THAT.
IT'S IN MULTIPLE WAYS, THE FIRST
IS NADH, PRODUCED IN THE PCA
INTO METABOLIC PATHWAYS.
THE SECOND IS FADH, WHICH IS THE
RESULT OF TWO.
SO WE CAN FUEL THIS PROCESS FROM
TWO DIFFERENT DIRECTIONS,
COMPLEX TWO OR COMPLEX ONE.
REGARDLESS OF WHICH WAY THAT
FUEL COMES IN, THERE'S THE
OPPORTUNITY FOR LEAKAGE IT THE
COMPLEX THREE.
THIS IS A LITTLE BIT MORE
UNUSUAL BUT A PHENOMENON KNOWN
AS REVERSE ELECTRON TRANSFER CAN
OCCUR.
THIS IS THE WRONG VERSION OF
THIS.
HOLD ON ONE SECOND.
CAN WE TAKE A TEN SECOND BREAK
AND LET ME SWAP OUT THE VERSE
HERE.
SORRY.
I ASSUME ONE OF YOU WAS GOING TO
NOTICE IT DIDN'T WATCH THE
HANDOUT.
LET ME SEE.
>> I APOLOGIZE FOR THAT.
I HAD TOO MANY VERSIONS ON HERE
AND I THINK I GOT THE WRONG ONE
BUT WE'LL MAKE DUE.
OKAY.
SORRY ABOUT THAT, BUT WE'LL MAKE
DUE.
THERE WILL JUST BE A LITTLE BIT
PROBLEMATIC FOR ME.
SO WE HAVE THE NADPH OXI DATES
SO ONE OF THE FUN THINGS THAT'S
COME OUT IN THE LITERATURE
RECENTLY, JUST TO MAKE SURE
YOU'RE ON THE CUTTING EDGE, IS
THESE ARE VERY TIGHTLY
CONNECTED.
THERE WAS A WONDERFUL STORY THAT
CAME OUT VERY RECENTLY THAT
SUGGESTED THAT ROS IS DIRECTLY
INVOLVED IN ELICITING
MITOCHONDRIAL ROS.
IF YOU THINK ABOUT IT, IT MAKES
PERFECT SENSE THAT TO PUSH THE
ROS PRODUCTION, GET EVERYTHING
INTO THE PHAGOSOME SO WE CAN
KILL THE CELLS.
I DON'T KNOW HOW MUCH YOU GUYS
HAVE HEARD ABOUT CHRYOSINE BUT
IT'S THE RECONFIGURATION OF THE
SUPER COMPLEXES.
YOU'RE FAMILIAR WITH THE ONE TO
FIVE OF THE TRANSPORT CHAIN.
WELL, THESE COMPLEXES ARE
CONFIGURED INTO SUPER COMPLEXES
WHERE, FOR EXAMPLE, YOU'LL GO
ONE THREE FOUR OR TWO THREE FOUR
OF THESE TOGETHER.
IN A FASCINATING STORY IT, WHAT
THESE AUTHORS WERE ABLE TO SHOW,
A KEY PROTEIN IN COMPLEX TWO AND
IT WOULD DRIVE A REORGANIZATION
OF THE SUPER COMPLEX TO SEE
FAVOR THE ACTIVITY OF COMPLEX
TWO.
THIS ALLOWED A LOT OF SUCCIIN
ATE AND BLOW OUT A LOT OF ROS IN
THE MITOCHONDRIA.
SO IT'S REALLY A VERY COOL
PATHWAY AND A NEAT MECHANISM BY
WHICH THESE REDOX SYSTEMS ARE
WORKING BY COLLABORATION TO GET
THE JOB DONE.
SO ONE OF THE OTHER REACTIVE
OXYGEN SPECIES OR REACTIVE
NITROGEN SPECIES IS N.O.
I KNOW YOU'VE HEARD A LOT ABOUT
NOS.
YOU'RE GOING TO HEAR EVEN MORE
ABOUT IT AT 5:00 I THINK.
RIGHT.
SO THE SINTHEY TKA*S TACE IS
HELD BY NADPH SHOWING US A
DIRECT METABOLIC CONNECTION.
IT GENERATES N.O.
THROUGH CHEMISTRY I WON'T GO
INTO PRODUCES THE EFFECTER
MONTHLY CULTURE THAT WILL CAUSE
OTHER REDOX LIKE SIGNALING, AND
THAT CHEMISTRY IS SHOWN DOWN
HERE WHERE THEY ARE SHOWING A
VARIATION ON IT WHICH IS THE
REACTION WITH THE N.O. WITH THE
SUPER OXIDE THAT MAY ALREADY BE
IN THE PHAGOSOME RESULTING IN
PROXYNITRITE AND WHETHER IT'S
THE EFFECTER MONTHLY -- MOLECUL.
OKAY.
SO LET'S LOOK A LITTLE BIT AT
METABOLISM.
SOME PEOPLE THINK ABOUT THIS
SORT OF THING OF THEY THINK
ABOUT METABOLISM AND IT IS
HORRIBLY COMPLICATED BUT IT'S
NOT THAT COMPLICATED I THINK.
THE ASPECTS I'M GOING TO SHOW
YOU TODAY ARE WHAT I HOPE I'M
GOING TO BE ABLE TO TIE IN.
HOPEFULLY THEY WILL BE ABLE TO
TIE IN NICELY FOR YOU TO WHAT
WE'VE BEEN DOING AND THE
CONNECTIONS IN REDOX.
I PUT THESE IN HERE SIMPLY SO
THAT YOUR HANDOUT HAS ONE FULL
PAGE.
NOW I KNOW I PRINT IT HAD SMALL
IN THERE.
WHAT WE'RE GOING TO DO IS ZOOM
IN ON THIS OVERALL VIEW AND I'M
GOING TO OUTLINE, FIRST OF ALL,
REFRESH YOUR MEMORY ON A COUPLE
OF THESE IN CASE YOU DON'T
REMEMBER YOUR BIOCHEMESTRY AND
WE'LL ZOOM IN ON A COUPLE OF
THEM IN A FEW STEPS AND HOW
THESE ARE TIED IN TO AN
INFLAMMATORY PROCESS.
SO THE FIRST ONE IS GLYCOLOSIS.
CELLS BECOME GLYCOLITIC.
IT DOESN'T AND AT LACTATE.
EVERYBODY SAYS THEY ARE
GLYCOLITIC.
THEY PROBABLY ARE BUT GLYCOLYSIS
IS PRIOR TO THAT.
IT'S HOW YOU CAN SEPARATE THE
WHEAT FROM THE CHAFF.
IT'S GLUCOSE FUELED.
IT'S VERY FAST.
YOU'VE MAYBE HEARD PEOPLE SAY IT
SWITCHES OVER TO GLYCOLYSIS.
YOU ONLY MAKE A LITTLE BIT OF
ATP.
WELL, OXPHOS IS SLOW.
THEY CAN ALSO KEEP THEIR REDOX
IN CHECK.
SO IF YOU MESS WITH THAT
GLYCOLYSIS, SO A VARIETY OF
CHANGES IN THE GLYCOLITIC SCHEME
WITH THE NADPH RATIOS AND
NECESSITATE OTHER PATHWAYS.
I THINK MAYBE WE'LL TALK A
LITTLE BIT ABOUT THOSE.
IT HAS A NET OF 4 ATP.
YOU START WITH SIX CARBONS, YOU
SPLIT INTO A DOUBLE OF THREE AND
YOU RUN THE BOTTOM PART TWICE
FOR THREE.
AND IT HAS A VALVE AT THE END.
IT HAS A RATE WITHERING STEPS
WHICH IS PKM.
IT STANDS FOR PROTEIN
KINASE MUSCLE AND IT GENERATES
ATP AND THIS IS A RATE
ELIMINATING STEP AND NOW THE
CELL IS GOING TO MAKE A
DECISION.
DO I PUMP THAT INTO THE
MITOCHONDRIA AND USE THOSE
CARBONS FOR OXIDATED
PHOSPHORILATION.
HOPEFULLY YOU'RE THINKING, YES,
THAT SOUNDS FAMILIAR FROM
BIOCHEMESTRY.
THE PENTOSE FOSS PATRIOT AND WE
THINK OF IT HAS A PATH FATE.
I WOULD ARGUE IN IMMUNOLOGY IS
NADPH.
SO THIS IS A RAPID, EFFICIENT
WAY FOR THE CELLS TO GENERATE
NADPH.
THIS IS A BIG SOURCE OF NADPH
FOR THE OXIDACE.
IF YOU LOOK AT THE OXIDATIVE
BURST, THE ABILITY TO GENERATE
OXIDATE SPECIES.
THIS IS DEPENDENT ON SUGAR.
BUT IT'S ALSO VERY IMPORTANT IN
BUILDING BLOCKS, AND SO THE
PENTOSE PHOSPHATE PATHWAY IS FOR
CELLS THAT HAVE TO MAKE OTHER
STUFF.
WHEN YOU THINK ABOUT METABOLISM,
YOU THINK ABOUT APP, RIGHT, THIS
IS THE MOST IMPORTANT.
NO, WHAT'S REALLY IMPORTANT IS
MAKING OTHER CELLS.
AND THE PENTOSE PHOSPHATE
PATHWAY AS WELL AS TRYPTOPHAN,
SISTIDINE AND BUILD OTHER CELLS.
OKAY.
SO THAT'S HOW I'D LIKE YOU TO
THINK ABOUT THE PENTOSE.
THE LAST ONE IN OUR LITTLE
REVIEW HERE IS THE TCA.
THERE'S THE TCA.
IT'S RUNNING CLOCK WAY FOR OX I
OXIDATION.
IT HAS A DESTINATION MAINLY OF
RUNNING INTO THE CYCLE OR IT CAN
BE BUILT IN CITRATE.
A COMPOUND WHICH IS QUITE A COOL
STORY AND WHICH YOU STIMULATE A
MACROPHAGE IS CONTINUES OUT
THAT'S DIRECTLY MICROBIALCITEAL.
I THINK IT'S A REALLY COOL
EXAMPLE OF EVOLUTION, KIND OF
PLAYING TENNIS SO THE BACTERIA
CHANGE THEIR IF I PHYSIOLOGY.
THE MACROPHAGE RESPONDS BY
SAYING IF YOU'RE GOING TO DO
THAT, I'M GOING TO KILL YOU
ANYWAY.
SO IT INCLUDES COMPLEX TWO OF
THE ELECTRON TRANSPORT CHANGE.
THEY TEND TO THINK OF THE
ELECTRON TRANSPORT CHAIN OF
BEING INDEPENDENT AND THE TCA
MAKES FUEL THAT WILL GO INTO THE
CHAIN AND THAT'S TRUE BUT THEY
ARE COMPLETELY INTEGRATED INTO
THE COMPLEX TWO WHICH IS
INVOLVED IN THE CONVERSION THAT
PERMITS THE CYCLE TO RUN.
IT'S FUELED BY PYRUVATE WHICH
CAN ENTER BY TWO DIFFERENT
ROUTES.
THERE'S A SECOND ROUTE THAT I
HAVEN'T DROWN IN HERE WHERE
PYRUVATE WOULD COME DOWN.
I'LL SHOW YOU SOME OF THE
SIGNIFICANCE OF THAT IN A LITTLE
BIT.
IT CAN BE FUELED ALSO BY
GLUTAMINE AND IT COMES IN AND
ENTERS INTO GLUTERATE.
WE'RE GOING TO TALK ABOUT TWO
KEY COMPONENTS IN HERE.
I THINK YOU'VE HEARD A FAIR
AMOUNT ABOUT THEM, PERHAPS IN
SOME OF THE OTHER PATHWAYS AND
I'M GOING TO HIT ON THOSE AGAIN
TO MAKE SURE THAT YOU FULLY GET
THAT CONNECTION.
THE LAST ONE.
ARGENINE IT GENERATES N.O., IT
CONSUMES NADPH.
SO AGAIN A FUEL THAT'S BUILT BY
METABOLISM IS CRITICAL FOR THE
PRODUCTION OF THIS REACTIVE
OXYGEN SPECIES THAT'S BEEN
INVOLVED IN THE BACTERIAL
CYTAL ACTIVITIES.
ARGINININE CAN REAGAIN RATE.
IT KAOEPLZ IT COMING WHICH SEATS
THE CYTRULLENE BEING MADE.
IN FACT WE HAVE A MOUSE MODEL
WHERE WE'RE NOW LOOKING AT THIS
IN DETAIL BECAUSE OUR TECHNICIAN
SAID THEY LOOK FINE BUT WHEN YOU
OPEN THEM UP, THEY STINK.
IT HAS MULTIPLE REDOX STEPS.
IT USES NADPH BY NOS.
IT CAN PRODUCE HYDROGEN
PEROXIDE.
THIS IS ANOTHER BIG REDOX STEP.
AS YOU BUILD POLYIAMINES, IF THE
CELL WANTS TO USE THOSE, IT WILL
USE THOSE FOR PROLIFERATION,
AMONG OTHER THINGS.
IF IT DOESN'T, THOSE CAN BE
OXIDIZED OUT AND PUSHED BACK
DOWN AND IN THAT PROCESS THEY
GENERATE HYDROGEN PEROXIDE SO
THERE'S A REDOX COST FOR THAT.
IF YOU DON'T USE THE POLYAMINES,
THERE'S A REDOX COST.
THERE'S AN EVIDENCE YOU MIGHT
WANT TO SPIT THOSE ON TO
SOMEBODY ELSE AND THEY WOULD
HAVE TO DEAL WITH IT AND THEY
WOULD PAY THE REDOX COST.
IN SOME MODELS POLYAMINES ARE
DETRIMENTAL.
YOU'LL SEE THE COMMON MADE
FUMARATE WHICH IS ALSO IN THE
TCA.
WHILE YOU'RE REGENERATING
ARGINONINE, AND IT'S COUNTER
REGULATED.
THERE'S COUNTER REGULATION
BETWEEN ARGINONACE AND NOS.
YOU CAN REGULATE N.O. BY THE
EXPRESSION OF ARGINNINE.
THERE'S A MECHANISM FOR
REPLENISHING THE ARGININE BY'SBY
ASPARTATE.
I'LL POINT OUT SOME OTHER
CHEMISTRY AS WE GO INTO THIS AND
ITS CONNECTIONS.
AS I MENTIONED THROUGHOUT, WE'RE
REALLY LOOKING AT A LOT OF
MYELOID CELLS.
THEY ALSO HAVE SOME VERY
SPECIFIC ACTIVITIES THAT ALLOW
CERTAIN SUB TYPES OF MACROPHAGES
TO FUNCTION A SPECIFIC WAY, AND
THEN THEY HAVE SPECIFIC
METABOLIC PATHWAYS AS I
MENTIONED THAT ARE INVOLVED IN
CYTOLITIC ACTIVITIES.
THERE ARE SUPPRESSIVE
MYELOID CELLS THAT ARE INVOLVED
WITH TUMORS SO THAT MAKES IT
INTERESTING TO GUYS LIKE US IN
THE CANCER INSTITUTE.
THAT'S NOT -- IT DOES INVOLVE
SOME REDOX BUT WE'RE NOT GOING
TO TALK ABOUT THAT TOO MUCH
TODAY.
LASTLY THERE'S IMMUNEO
SUPPRESSION.
THIS ISN'T LASTLY BUT THERE'S
IMMUNEO SUPPRESSION.
INTERESTINGLY THOSE ARE REDOX OR
REDOX RELATED PROTEINS.
OF COURSE THEY ARE DRUGGABLE.
THAT'S ALWAYS FUN.
THAT'S A GOOD THING WHEN YOU'RE
TRYING TO HELP PEOPLE IS TO WORK
OUT PATHWAYS THAT ARE DRUGGABLE,
AND THEN YOU CAN DOUBLE HIT.
SOME OF THEM ARE IN THE PAM WAY
CELLS, SOME ARE IN THE IMMUNEO
CELLS.
AND SO A LOT OF PEOPLE HAVE BEEN
LOOKING FOR THAT SORT OF
ACTIVITY.
AND LASTLY THEY EVEN DO IT, YOU
KNOW, ON THE BIG BANG THEORY.
SO IF THE BIG BANG THEORY IS
DOING METABOLISM, THEN YOU
SHOULD TOO, IS THE WAY WE LOOK
AT IT.
WHAT'S HAPPENING IN THESE
METABOLIC SWITCHES WITHIN THE
IMMUNE SYSTEM.
SO THE TRADITIONAL VIEW OF
METABOLISM IS THAT IN THE
PRESENCE OF OXYGEN AS WE JUST
REVIEWED, GOOD THE ALTERNATIVE
IS IN
THE ABSENCE OF OXYGEN,
GLYCOLYSIS TURNED INTO PYRUVATE
AND MAKES LESS.
CELLS WILL RUN THIS PATHWAY EVEN
IN THE PRESENCE OF OXYGEN.
REMEMBER AT FIRST IT WAS A BIT
OF A CONUNDRUM.
WELL, THE REASON WE DO THAT IS
BECAUSE YOU CAN MAKE IT VERY
FAST AND YOU GENERATE A LOT OF
LACTATE.
LACTATE CAN BE SPIT OUT AND
PEOPLE HAVE NOW BEEN VERY
INTERESTED IN WHY TUMOR CELLS DO
THIS BUT OTHER PROLIFERAITIVE
TISSUES DO THIS.
SO IT WAS A LITTLE BIT OFF ON
THAT BUT IT LOOKS LIKE IT'S
PRETTY IMPORTANT.
OTTO ALSO WORKED ON LEUCO
CYTESIN 1956.
THREE'S THESE MACROPHAGES.
WE'RE GOING TO SWITCH THEM.
WE'LL TAKE THEM OUT OF THE
RESTING STATE AND PUT THEM UNA
RESTING STATE.
AS I MENTIONED BEFORE IS THEY
MOVE TOWARD GLYCOLOSIS.
REMEMBER WHEN LACTATE LEAVES THE
CELL, THE LACTATE TRANSPORTERS
TRANSPORT LACTATE PLUS A PROTON.
AS THE LACTATE GOES OUT, THE
MEDIA GETS ACID D. --
ACIDIIFIED.
THEY PRODUCE A LOT MORE LACTATE.
IF YOU GRAPH IT, IT LOOKS LIKE
THIS.
WE LOOK AT IT THIS WAY WHERE WE
HAVE CELLS IN THESE RESTING
STATE.
WE DON'T REALLY LIKE THOSE TERMS
BUT THIS IS SO-CALLED GLYCOLITIC
COMMITMENT.
WE CALL IT COMMITMENT BECAUSE
THEY ACTUALLY TURN OFF THEIR
OXIDATIVE PHOLITION.
YOU CAN SEE HOW MUCH IS REALLY
CONSUMED BY THE ELECTRON
TRANSPORT CHAIN, THEN YOU CAN
DECOUPLE THE MITOCHONDRIA, AND
SEE IF I REALLY LET THEM RUN
HARD, HOW MUCH OXYGEN WILL THEY
CONSUME, THAT'S THEIR MAXIMUM
RATE AND WE CAN POISE IT ALL THE
WAY TO GET TO A BASE LINE.
AN LPS STIMULATED CELL IS PRETTY
FLAT.
SO IT'S TURNED OFF ALL OF IT
MITOCHONDRIALY DERIVED AND NOW
THIS LITTLE SWITCH MAKES A
LITTLE BIT MORE TENSE TO YOU I
HOPE.
SO THEY'VE SWITCHED FROM AN
OXIDATIVE SITUATION TO A
GLYCOLITIC SITUATION.
YOU ARE COMMITTED TO GLYCOLOSIS.
YOU'RE COMMITTED TO SUGAR.
THERE'S A COUPLE OF KEY
SUBSTRATES THAT I WANT YOU TO
THINK ABOUT.
THIS IS UNBIASED METABOLOMIC.
RED SHOWS WE SET THE RESTING TO
THE WHITE, RED GOING UP, BLUE
IT'S GOING DOWN.
SO THERE'S AN INCREASE IN ALL
THESE SUBSTRATES AND AN INCREASE
IN LACK TICK ACID.
ONE OF THE MOST INTERESTING
THINGS I WANT TO POINT OUT,
WE'RE GOING TO COME BACK TO THAT
IN JUST A MINUTE.
THIS HAD BEEN -- WE'LL COME BACK
TO IT NOW.
THIS HAS BEEN TURNED A BRICK.
AS CARBON RUNS THROUGH THE TCA,
IT SEEMS TO BREAK.
I'LL SHOW YOU A LITTLE BIT MORE
ABOUT WHY THAT'S A BREAK IN A
SECOND.
WE KNOW THE KEY CELLS MAKE N.O.
A RESTING CELL HAS A DRAMATIC
INCREASE IN CYTROLINE DURING THE
PRODUCTION OF N.O. WHERE AS
ORNOTHINE-- SOME OF THAT GETS
CONVERTED OFF TO POLYAMINES.
THE PATHWAYS I JUST SHOWED YOU,
YOU CAN KIND OF SEE WORKING AS
WE STIMULATE THE CELLS.
SO THIS IS WHAT IT LOOKS LIKE
OVERLAID.
WITH US THERE'S AN INCHRIS IN
CITRATE.
IT IS THE ITACONATE LEVEL.
THIS PLAYS A PART IN THAT
OXIDATIVE GOING OFF.
ITACONATE WOULD COME ON AND CAN
SUPPRESS COMPLEX TWO.
I WANT YOU TO PAY ATTENTION TO
THIS LITTLE RATIO.
AL PA -- ALPHA KETOGLUT ARATE.
SO WHY IS IT SO IMPORTANT?
THE REASON IT'S SO IMPORTANT IS
THEY ARE CO-FACTORS FOR A
VARIETY OF ENZYMES WITHIN THE
CELL.
ONE OF THE MOST IMPORTANT ONES
ARE THE PDHs WHICH I KNOW DAVE
IF I THINK TALKED TO YOU ABOUT.
USE OXO GLUTTERATE WITH
MOLECULAR OXYGEN TO PRODUCE HIGH
ANTHRAX -- HIGH DROXILATEED
PROTEIN.
WHAT YOU CAN IMAGINE IS IF THE
SUCCINATE LEVELS GO HIGH, WHAT
WILL HAPPEN TO THIS REACTION, IT
WILL SLOW OR STOP AND THAT MEANS
THAT THE ABILITY OF THE PHDs
TO HIGH DROXILATE GO DOWN.
THEY ARE TKROLD BY REDOX.
THEY ARE AN IRON BASED ENZYME
SYSTEM.
THEIR REDOX STATE IS CRITICAL SO
ROS PRODUCTION CAN SHUT THEM
DOWN.
I MENTION JUST ANOTHER INHIBITER
CAN SHUT THEM DOWN.
HYPOXIA CAN SHUT THEM DOWN
BECAUSE THEY USE MOLECULAR
OXYGEN.
THEY SENSE A LOSS OF OXYGEN BUT
THEY CAN BE REGULATED BY REDOX
AND BY THE SUBSTRATES AS WELL.
SO WHAT HAPPENED WHEN THEY ARE
REGULATED, AND I THINK THIS WAS
TOUCHED ON BY A COUPLE OF PEOPLE
INCLUDING TERRY WHEN HE TALKED
TO YOU.
SO WHAT HAPPENS IS THEY
STABILIZE THE HIP.
AGAIN, THEY ARE CONVERTING ALPHA
KETOGLUTINATE.
SO YOU CAN HAVE IT MADE
CONTINUOUSLY BUT YOU DON'T SEE
IT IN THE CELL BECAUSE IT'S
TURNED OVER VERY QUICKLY.
WHAT HAPPENS WHEN THESE RATIOS
WOULD CHANGE.
THE SUCCINATE IS VERY HIGH AND
THIS IS KNOW AS PSEUDO HYPOXIA.
THE PHD IS GOING OFF EVEN THOUGH
MOLECULAR OXYGEN IS AVAILABLE.
WE'RE THINKING THAT THE PHD GOES
OFF DO YOU SEE THE SUCCINATE
IMBALANCE.
IS THAT WHAT'S HAPPENING?
WE THINK IT IS IT.
I SHOWED YOU ALL THOSE RED
SUBSTRATES WHEN THE CELLS ARE
STIMULATED?
IT TUPBZ OUT ALL OF THESE
ENZYMES, EVERYTHING THAT'S IN
PINK IS A TARGET OF HIF.
IT'S A TARGET OF THE REGULATION
BY THE PRO HYDROXILATE SO THAT
THE SUGAR CAN CONTINUE TO FLOW
IN AND/OR BE PASSED OFF TO PENT
PENTOPHOSPHATE.
WHAT THAT KINASE DOES, THE FIRST
STEP IN THE PRODUCTION OF
CYTRATE.
OKAY.
SO CAN WE SEE THIS PROCESS
ACTUALLY WORK?
IT'S KIND OF FUN BECAUSE YOU
CAN.
YOU CAN TAKE LABEL GLUCOSE AND
DUMP IT INTO CELLS.
YOU CAN TRACK THE CARBON
THROUGH.
YOU CAN SAY IF I ADD GLUCOSE
THAT'S ALL HEAVY, THE PYRUVATE
WILL BE ALL HEAVY.
THOSE TWO CARBONS WILL COMBINE
WITH FOUR AND MAKE A SIX CARBON
CITRATE.
WE CAN ASK IS THE CITRATE TWO
CARBONS OR FOUR HEAVIER THAN IT
SHOULD BE AND THAT ALLOWS US TO
WATCH THE FLUX OF CARBONS.
INITIALLY THERE IS SOME PLUS TWO
CITRATE.
SO THE SUGAR IS BEING MADE INTO
CIT DISRATE AND TO SEE TOO
HEAVY.
COMES ON, SHUTS OFF THIS VALVE.
THE PATHWAY IS NOW SHUT OFF AND
THERE IT IS, YOU SEE IT GOING
ON.
YOU LOOK AROUND THE REST OF THE
CYCLE.
YOU CAN ONLY SEE THE PLUS TWO.
YOU DON'T SEE ANY OF THE PLUS
THREE OR YOU DON'T SEE ANY OF
THE PLUS TWO AFTER INTERFERON
BECAUSE IT'S BEEN SHUT OFF AT
THE SOURCE.
I TOLD YOU THAT THOSE RATIOS
CHANGE TOO.
HERE IT IS.
IF I LOOK AT THE RATIO OF
CITRATE, THERE'S 14 TIMES MORE
LABELED CITRATE AN ALPHA CAN HE
TELLOGLUTTA RATE.
AND WE THINK THAT'S WHY THE
LEVEL FALLS TO START TO SET UP
THAT DIFFERENTIAL THAT WILL TURN
OFF THE PRO HYDROXILATEERS.
THAT BYPASSES PYRUVATE AND
ENTERS ANOTHER WAY.
IT STICKS A CARBON ON THE END SO
THIS IS NOW FOUR LABELED
CARBONS.
BY LOOKING AT THE PLUS THREE
CITRATE, WE CAN SEE THAT THE
SWITCH HAS OCCURRED.
SO THE CELL CHANGES THE ROUTE OF
CARBON GOING IN.
YOU CAN FOLLOW THAT THEN ALL THE
WAY AROUND THE CYCLE SUGGESTING
THE CARBON IS ENTERING A
DIFFERENT WAY.
IT'S GOING OUT TO ASPARTATE AND
IT'S DRIVEN BY PYRUVATE.
OTHER PEOPLE SEE THE SAME THING
AND THEY SAY THIS IS BECAUSE OF
A BREAKDOWN IN ADH, AND THAT'S
WHY YOU GET THIS CHANGE IN RATIO
AND WE THINK THAT'S WRONG AND
I'LL SHOW YOU WHY IN JUST A
SECOND.
SOME QUICK PRIMARY DATA THAT
I'LL GO THROUGH VERY QUICKLY
BECAUSE IT'S NOT THE BIG GOAL OF
WHAT WE WANT TO DO HERE.
WHAT WE FIND IS REGULATORY
CYTOKIN, IF YOU INTERFERE WITH
THE USE OF SUGAR, YOU GET LESS.
IF YOU STIMULATE THEM IN
ELIMINATING SUGAR, WE CAN MAP
OUT THE MECHANISM AND SHOW THAT
AS ATP SYNTHESIS THAT FALLS,
COMES ON, SUPPRESSES TORQUE AND
PRESENTS IL-10.
SO WE BUILT OURSELVES A LITTLE
MODEL BECAUSE IT'S ALWAYS FUN TO
BUILD MODELS, THAT WHEN YOU
STIMULATE YOU MAKE 10s, IT
REGULATES BACK AND THAT SHOULD
REGULATE THE COMMITMENT AND SO
IT DOES.
THEY OVERSHOOT.
THEY GO SO FAR INTO FLY --
GLYCOLOSIS THEY ARE ALMOST GOING
OFF THE CLIFF.
SO WHAT WE NOTICE IS WHEN YOU
STIMULATE CELLS AND YOU CAN LOOK
AT THEIR OXIDATIVE GOING OFF,
SOME THING WORK LIKE THIS, SOME
THINGS DON'T, AND IT TURNS OUT
THAT THAT'S IN DIRECT CORE PLACE
TO THIS ABILITY TO PRODUCE N.O.
IF YOU CAN PRODUCE N.O., YOU
CRUSH THE OCR.
IS IT N.O., LOOKS LIKE IT IS?
YOU CAN ADD IT DIRECTLY AND
CRUSH THE OCR.
THAT WHOLE COMMIT WE THINK IS
DUE TO THE REDOX ACTIVITY OF THE
N.O.
HERE IT IS, IS NO, HITTING IN
COMPLEX ONE HAVE BEEN SHOWN IN A
MECHANISM.
BECAUSE THEY CAN COMPETE WITH
SITES, THEY CAN INTERFERE WITH
COMPLEX FOUR.
BOTH OF THOSE WOULD ALLOW THE
N.O. TO SHUT IT OFF.
LET'S SEE.
IF WE ADD THE OXPHOSE GOES OFF,
THAT'S GOOD.
SO IT'S ENTIRELY DEPENDENT ON
THE N.O., AND IN FACT WE CAN
SHOW THAT THE IL-10 REGULATES
THAT BECAUSE IL-10 IN THOSE
CELLS MAKE MORE N.O.
SO THE CELL, AND I THINK I
SUMMARIZE IT HERE, SO THE CELL
IS REGULATING GLYCOLAITIVE
COMMITMENT.
OKAY.
SO THEN IF WE KNOCK OUT N.O.
THROUGH INOS.
THEY CAN STILL REGULATE
GLYCOLOSIS.
THE LEVELS DON'T FALL.
THERE'S STILL A DIFFERENCE HERE
BUT IT'S NOT AS SHARP AS IT IS
HERE.
WE KNOW OUR EXPERIMENT WORKS
BECAUSE THE CITRULLINE DOESN'T
WORK.
WE THANK GOODNESS WE PUT THE
RIGHT MICE IN THERE.
IF WE LOOK AT THE CARBONS, WHICH
ARE IN THE WARM COLORS HERE.
WHAT HAPPENS IS YOU DON'T SEE
THE M2 GO OFF, YOU DON'T SEE THE
VALVE CLOSE AND DON'T SEE THE M3
COME ON.
SURE ENOUGH FORECLOSURE LOOK AT
THE GENES, YOU'LL SEE HERE, THIS
IS THE UP REGULATION THAT SHUTS
IT OFF, DOESN'T COME ON.
HERE'S THE UP REGULATION OF THE
ENZYME THAT BYPASSES, DOESN'T
COME ON IN THE NOS.
HERE IS THE HIF TARGET.
IT'S TELLING US THEY ARE NOT
FUNCTIONING PROPERLY TO TURN
PDK1 ON.
SO IT WAS A BIG CLUE TO US.
IF THEY ARE NOT FUNCTIONING
PROPERLY AND IF THEY ARE N.O.
DEPENDENT, HOW IS THE
OXFOS GOING OFF.
YOU LOOK AT IDH1 AND YOU THINK
ABOUT REDOX AND DAVE SHOWED YOU
A BUNCH OF DIFFERENT WAY THAT'S
N.O., FOR EXAMPLE, CAN HIT.
BUT THAT DOESN'T WORK VERY WELL
ON IDH1.
THERE'S NO INDICATION IT SHOULD
WORK.
WHOSE WRITING A PAPER SAYING IT
CONTROLS THE TCA IN
MITOCHONDRIA.
I DON'T THINK THEY WERE THINKING
ABOUT REDOX BIOLOGY.
WE WERE.
WE SAID THERE'S THIS OTHER
ENZYME AND IT'S INVOLVED IN THAT
SAME SORT OF STEP.
SO IF N.O. IS REQUIRED FOR THIS
LITTLE BREAK TO OCCUR, RIGHT,
THEN N.O., IF IT'S IDH1, N.O.
SHOULD BE REQUIRED FOR THAT.
IT GOES DOWN EVEN WHEN YOU BLOCK
N.O.
THEY GOT IT WRONG.
THEY DON'T KNOW IT YET BECAUSE
WE'RE JUST WRITING IT UP BUT
THEY ARE GOING TO FIND OUT SOON
IF THEY ARE WRONG.
SO -- I'LL EVEN SKIP THAT
BECAUSE I DON'T WANT TO RUN OUT
OF TIME.
WE'RE ALREADY RUNNING A LITTLE
BEHIND.
A MUCH BETTER TARGET.
I THINK DAVE SHOWED YOU SOME OF
THE CHEMISTRY AS TO HOW N.O. AND
OXYGEN RADICALS WOULD HIT
ACONOTASE WORKS.
IF YOU REMEMBER, THIS IS THE
THING THAT'S CONVERTED INTO
ITOCONNICK ACID, IT SUCKS IT OUT
OF THE MIDDLE.
IT'S AN IRON SULFUR HUGE, SO
IT'S A CUBE STRUCTURE AND IT'S
VERY SUBJECT TO REDOX ATTACK AND
ACTUALLY THERE'S OLD STUFF IN
THE LITERATURE THAT SHOWS THAT
N.O.
OXIDIZE ONE OF THESE IRONS AND
KICK IT OUT OF THERE AND DESTROY
THE CUBE.
IN FACT YOU CAN GO BACK, ADD IN
IRON AND REDUCING AGENTS AND FIX
IT AND PUT THE IRON BACK IN.
SO IT WAS A REALLY GOOD TARGET
AND IN FACT WHEN WE LOOKED, WE
FIND THAT THE STIMULATION TURNS
OFF ACTIVITY AND THAT REQUIRES
N.O.
SO IF WE BLOCK IT, IT ALL GOES
AWAY.
SO IDH1 IS NOT THE TARGET.
THIS IS REDOX BIOLOGY HITTING
ACONIIT'S A.
THE BLACK LINE HERE, CAN DO OX.
IT CAN STATE THAT CITRATE,
GENERATE COMPLEX ONE AND I'LL
GET OXYGEN CONSUMPTION.
THERE'S A PLUMMET SO THEY CAN'T
USE CITRATE.
THERE'S THIS BREAK BETWEEN
CITRATE AND ALPHA KETO
KETOGLUTTERATE.
THE ONLY ENZYME PATHWAY THAT I'M
SKIPPING IS ACONITASE.
SO IF I BYPASS ACONITASE BY
GIVING THE BYPRODUCT OF
ACONITASE, THEN THEY WORK FINE.
IT DIDN'T MESS WITH THOSE.
WHAT IT DID IS IT STOPPED
ACONITASE AND THAT'S HOW IT'S
TURNING OFF THE PATHWAY.
WE CAN LOOK AT THAT VERY
SPECIFICALLY AND OF COURSE IN
THE NOS IT DOESN'T WORK.
THEY WILL ACCUMULATE LIPIDS TOO.
I SHOULDN'T HAVE PUT THAT IN
THERE SO WE'LL SKIP IT.
I WANTED TO COME BACK HERE TO
REMIND YOU THAT MITOCHONDRIA
DERIFED ROS IS ABSOLUTELY A BIG
DEAL HERE AND CAN STILL CONTROL
THE METABOLIC ACTIVITY BECAUSE
THIS OROS STARTS TO HURT THE
MITOCHONDRIA, AND ONE OF THE
LAST THINGS I'LL SHOW YOU HERE
IN THE ACTUAL REDOX BIOLOGY IS
THAT THE INFLAMMATORY CYTOKIN
CYTOKINE 1, ONCE WE MESS WITH
THE FUNCTION, ROS PRODUCTION OUT
OF THE MITOCHONDRIA LEADS TO
OXIDATION OF MITOCHONDRIAL DNA.
THIS ACTIVATES A MULTI-PROTEIN
COMPLEX CALLED THE INFLAMASOME.
THERE'S A PRO CYTOKINE MADE.
AND IT'S CLEAVED INTO MATURE
IL-1 WHICH IS SECRETED.
IT'S TIED DIRECTLY TO THE REDOX
BIOLOGY AND THE CONTROL OF
MITOCHONDRIAL FUNCTION.
THE LAST THING I SHOULD SHOW
YOU, I SHOULD SKIP OVER THIS
BECAUSE I'M LATE BUT GENE
REGULATION IS NOT SO AFFECTED
BECAUSE OF IL-1.
I APOLOGIZE FOR PUTTING THE
WRONG TALK ON HERE.
I HAD SOME GREAT SUMMARY THINGS
AT THE END.
I'LL LEAVE IT WITH THIS, WHICH
IS A LITTLE BUSY.
THIS IS KIND OF AN N.O.
CENTRIC VIEW, IT CAN SUE DRESS
ACONITASE TWO WHICH IS
CONTROLLING THE CONVERSIONS
BETWEEN CITRATE AND ALPHA KETO
TKPWHR*S GLUTTER ATE SO THEY
FALL.
THE PHDs GO OFF, HIF ACTIVITY
GOES UP.
IT DRIVES, AS I SHOWED YOU, THE
KINASE, WHICH TURN OUT -- THE
ARGININE SO IT RUNS IN A NICE
CLEAN CIRCLE.
AT THE SAME TIME BEFORE THIS
ACONITASE GOES OFF, THAT CITRATE
THAT GOES OUT IS SEQUESTERED
INTO THE PRODUCTION OF ITA
CONATE AND THE WHOLE THING IS
REGULATED BY IL-10.
THERE'S OTHER METABOLIC CONTROLS
IN IMMUNITY OBVIOUSLY, AND CAN
YOU READ ABOUT THOSE.
THE REGULATION OF GLUCOSE
REGULAR ORGANIZATION.
IT IS MORE CLEARLY A REDOX
ACTIVITY, THAT'S WHY I PICKED ON
IT.
I WOULD RECOMMEND THAT YOU GRAB
UP ONE OF THESE REVIEWS AND
YOU'LL SEE A LITTLE BIT MORE IN
THE CONNECTIONS THERE.
JUST A COUPLE OF FINAL THOUGHTS
THEN, THE REDOX BIOLOGY TOUCHES
LOTS OF ACTIVITY IN THE CELLS.
THE ACTIVITY SHOULD BE MATCHED
TO A SPECIFIC MIX.
LOTS OF CELL BIOLOGY IS DONE THE
WRONG WAY.
RIGHT?
I REMEMBER GOING INTO A TALK
SEVERAL YEARS AGO.
A GUY WAS LIKE WHY ARE WE
STUDYING HUMAN CANCER IN A DISH
WITH 21% OXYGEN, RIGHT, TONS OF
SUGAR AND COW SERUM.
HOW FAR FROM REALITY CAN YOU
GET?
SO WHEN YOU THINK OF MET
METABOLISM,
YOU HAVE TO THINK OF WHAT'S
AVAILABLE TO THE CELLS.
EFFORTS ARE UNDER WAY TO MAKE A
MATCH, THOSE NICHES ARE FIXED,
AND MANY OF THOSE WILL INDUCE
REDOX MOLECULES LIKE N.O. OR THE
REGULATED PYRUVATE.
I'LL ALSO POINT OUT THAT MOST
THERAPIES THAT WORK UTILIZE
INTERFERON AND IT IS ALSO A GOOD
DRIVER OF MITOCHONDRIAL ROS.
YOU MAYBE HEARD ABOUT THIS, SOME
OTHER SPEAKERS ALREADY IN THE
COURSE.
BUT N.O. PRODUCTION IS
DRAMATICALLY DIFFERENT BETWEEN
MOUSE AND HUMAN.
AND IF YOU GO AND YOU LOOK AT
HOW CANCER THERAPIES ARE TESTED,
THEY ARE TESTED IN MICE.
SO SHOULD WE NOT BE SURPRISED
THIS THEY DON'T WORK WHEN WE GO
TO HUMAN BEINGS?
I THINK WHAT WE'RE FINDING IS
THAT ALL OF THESE METABOLIC
CHANGES THAT ARE BASED ON THAT
REDOX PATHWAY ARE GOING TO BE
DIFFERENT IN MICE AND HUMANS.
WE NEED TO DO IT.
I TAKE CREDIT FOR LOTS OF FUN
PEOPLE UP INLY LAB.
I SHOWED YOU WHAT'S DONE BY
ERIKA.
LUKE HAS DONE A LOT OF OUR
METABOLIC WORK AS WELL.
SOME OF THE IL-10 STUFF WAS DONE
BY WOLF BUT WE FIRED HIM BECAUSE
HE DRINKS TOO MUCH AND THIS IS
THE CURRENT GROUP.
I APOLOGIZE FOR GOING OVER A
LITTLE BIT AND WORST OF ALL
HAVING THE WRONG SLIDES FOR YOU.
I'M HAPPY TO CLARIFY ANYTHING I
CAN FOR YOU.
THANKS.
[INAUDIBLE]
>> IT'S HIGHLY INFUSIBLE.
SO ONE EIGHT OR TEN HOURS YOU'LL
SEE VERY HIGH LEVELS.
WE'VE ACTUALLY BEEN TRYING TO
LOOK NOW TO SAY IS THIS
PHYSIOLOGICALLY RELEVANT.
IN THE DISH, THERE'S A LOT OF
CELLS IN THE DISH.
WE CAN CONTROL THEIR DENSITY WE
THINK.
HAVE EVIDENCE IN DIVO, EVEN IN
HUMAN BEINGS THAT THERE'S
EVIDENCE OF N.O. PRODUCTION.
SO YOU CAN SEE EFFECTS OF N.O.
YOU CAN SEE THE CHEMICAL
BYPRODUCTS AND THE TISSUE SO YOU
KNOW THAT THE LEVELS HAVE GOTTEN
PRETTY HIGH BUT IT COMES ON
EXTRAORDINARILY FAST.
SOME OF THE EFFECTS ARE CELL
DENSITY DEPENDENT AND WE THINK
THAT'S BECAUSE AS AGAIN YOU'VE
HEARD FROM DAVE ALREADY, THERE'S
DIFFERENT CHEMISTRY FOR N.O.
GIVEN THE CONCENTRATION.
WHEN YOU PUT THEM TOGETHER IN
HIGH DENSITIES, YOU GET HIGH
LEVELS OF N.O. AND CERTAIN
CHEMISTRIES YOU WOULDN'T GET AT
LOW LEVELS, IF THAT MAKES SENSE.
THERE'S LITTLE KNOWN ABOUT HOW
IT GOES OFF.
WE'RE NOW LOOKING AT HOW THE
MESSAGE GOES OFF AND HOW
TRANSIENT THAT MAY BE NOW THAT
WE KNOW THAT A ACONITASE WILL BE
HIF.
WE FIND IT HAS TO BE REPLACED.
IT HAS TO BUILD NEW ACONITASE.
SO THE MECHANISM BY WHICH IT HIT
AND THE CONTINUING PRODUCTION IS
GOING TO BE IMPORTANT I THINK.
OKAY.
>> WE HAVE ONE ANNOUNCEMENT,
THAT BEING NEXT WEEK WE HAVE A
FINAL CLASS.
I'M WORKING ON A FINAL
EXAMINATION.
IT WILL BE POSTED ON THE WEBSITE
AND THERE WILL BE ONE QUESTION
FROM EACH LECTURE, AND IF YOU
GET 70%, YOU PASS AND THEN YOU
GET YOUR CERTIFICATE.
SO IT WILL BE -- THE EXAM WILL
BE POSTED FOR ABOUT A MONTH AND
A HALF.
SO YOU'LL HAVE PLENTY OF TIME TO
TAKE IT.
AND IT'S OPEN BOOK, OPEN NOTES.
IF YOU HAVE YOUR HAND-OUTS, YOU
SHOULD DO JUST FINE.
OKAY.
SO OUR LAST SPEAKER TODAY GOT
HER PHD AT THE UNIVERSITY OF
ARIZONA, GENTLY SHE WAS A POST
DOC AT THE UNIVERSITY OF SAN
FRANCISCO AND SHE'S BEEN AT
MCI NOW FOR TWO YEARS.
>> I'M A RESEARCH FELLOW IN
DR. DAVID BANKS' LAB.
I'M GOING TO TALK ABOUT THE
BIOLOGY OF TWO SMALL MOLECULES.
I'M SURE YOU HAVE HEARD A LOT
ABOUT NITRIC OXIDE.
IT IS DISCOVERED BY JOHN PRESLEY
IN 1-- 1972 BUT INITIALLY IT WAS
STUDIED IN TERMS OF IN
TOXICOLOGY BECAUSE AT THAT TIME
MIGHT TRICK OXIDE WAS WERE WERE
FOUND AS AIR POLLUTANTS.
AS A CARCINOGENIC COMPOUND IN
STORED FOOD.
SO MOST OF THAT PEOPLE STUDIED
THE HEALTH EFFECTS OF THOSE
TOXIC COMPOUNDS.
IT WAS NOT UNTIL 1998 NOBLE
PRIZE WAS GIVEN IN THE FIELD OF
NITRIC OXIDE, SO IT WAS SHOWN IN
LATE 1980s THAT NITRIC OXIDE
WAS COMBINED WITH -- SO IT'S
TWO-STEP REACTION WHERE FIRST
CONVERTED INTO --
THERE ARE THREE DIFFERENT IO
FORMS.
NO.
STWO IS THE INDUCIBLE ICEO FORM.
CAN ALSO BE PRODUCED IN A WAY
FROM NITRATE.
AS YOU CAN SEE HERE, NOs
ENZYME -- NITRATE CAN BECOME THE
MAIN SOURCE OF PRODUCTION OF
NITRITE.
SO THESE ARE -- THESE ARE SOME
OF THE REACTIONS OF NITRITES.
SO THE REACTION WHICH -- SO
THESE ARE SOME OF THE DIRECT
REACTION FROM NITRIC OXIDE
REACTS DIRECTLY.
APART FROM THAT NITRIC OXIDE CAN
ALSO REACT WITH OXYGEN TO
PRODUCE SOME OF THESE MACHINES
WHICH DAN ALREADY TALKED ABOUT.
WHILE N203 CAN LEAD TO NITRIC
ATION.
IS ONE OF THE MONTHLY HE CAN
HRAR OF THIS STEP AND THEN TWO
AMINO ACIDS TEND TO GO -- SO AS
OF THE DIRECT REACTION, ONE OF
THE IMPORTANT TARGETS OF NITRIC
OXIDE ALSO KNOWN AS FGC.
SO FTC IS A PROTEIN WITH AN IRON
TWO OXIDATION STATE WITH A
HISTAMINE RESIDUE AT THE BOTTOM.
SO WHEN NITRIC OXIDE BIND, THES
ARE DUE COMES OFF AND THIS
CAUSES CHANGE IN THE STRUCTURE
OF THE ENZYME.
SO THIS CHANGE INCREASES THE
ACTIVITY BY LIKE 200-FOLD.
AND THIS ENZYME THEN CONVERTS
GTP INTO A SECOND MESSENGER
MONTHLY CULTURE WHICH IS
RESPONSIBLE FOR MANY OF THE
PROPERTIES OF NITRIC OXIDE SUCH
AS MANY OF THE CARDIOVASCULAR
FUNCTION, AND MUSCLE RELAXATION
AND LIKE MANY OF THE OTHER
PROPERTIES UNDER PHYSIOLOGICAL
CONDITION.
NITRIC OXIDE ROLE IN
CARDIOVASCULAR FUNCTION HAS BEEN
SHOWN FOR A LONG TIME, BUT SINCE
IT'S A RADICAL MONTHLY CULTURE,
IT CAN ALSO REACT WITH OTHER
RADICALS LEADING TO CHAIN
REACTION, FORMING OUR N.O.
SPECIES.
IT ALSO PLAYS AN IMPORTANT ROLE
IN THE SYSTEM UNDER DIFFERENT
PATHOLOGICAL OR MICROBIAL
ATTACKS.
IN OUR LABS WE ARE MAINLY
INTERESTED IN THE CANCER BIOLOGY
OF NITRIC OXIDE WHICH I'M SURE
LISA TALKED QUIT A LOT IN THE
LAST CLASS.
SO THEN THE OTHER MONTHLY
CULTURE THIS I'M GOING TO TALK
ABOUT IS HNO.
SO HNO IS A REDUCED FORM OF
NITRIC OXIDE.
HOWEVER, IT HAS NOT BEEN
CONVINCINGLY SHOWN TO BE
PRODUCED UNDER PHYSIOLOGICAL
CONDITIONS.
THERE ARE MANY REPORTS WHERE
PEOPLE SAY THIS THEY HAVE
EVIDENCE FOR FORMATION OF UNDER
PHYSIOLOGICAL CONDITIONS.
ONE OF THE LIMITATIONS FOR
DETECTION IS IT HAS A REALLY
FAST SELF -- RATE.
THESE ARE SOME OF THE REACTIONS
WHICH ARE REPORTED IN THE LIT
ARE TOUR FOR EVIDENCE OF
FORMATION OF HNO.
SO ONE OF THE KEY PARTS BY WHICH
HNO CAN FORM IS FROM THIS
INTERMEET OF NOS REACTION.
ALSO IT CAN FORM A TRUE NOS WHEN
THERE'S A CO-FACTOR FOR THIS
ENZYME.
OTHER THAN THAT, HYDROXYREA CAN
ALSO UNDERGO OX IIDATION
REACTION
TO FORM HNO.
STUDIES HAVE SHOWN A SMALL
ETHANOL MONTHLY CULTURE WHICH IS
HSNO CAN REACT WITH H2 S WHICH
IS TRANSMITTER.
HNO MAINLY REACTS WITH
THIOL SPECIES.
SO THE REACTION OF HNO IS KNOWN
AS NIGHT ROWSYLATION.
THIS REACTION WHICH IS REACTION
OF HNO.
AS I MENTIONED EARLIER, HNO CAN
ALSO UNDERGO SELF -- TO BE USED
FOR DETECTION OF HNO.
SO DETECTION OF N2O OF
CONFIRMATION OF PRODUCTION OF
HNO.
OTHER THAN THAT, HNO CAN ALSO
FORM, CAN REACT WITH OXYGEN TO
FORM HNO OXYGEN INTERMEDIATE.
THIS MONTHLY CULTURE HAS NOT
BEEN CONVINCED.
SOME PEOPLE BELIEVE THAT IT IS
SIMILAR TO -- SPECIES.
THERE ARE SOME DIFFERENCES
BETWEEN THESE SPECIES.
CAN ONLY CAUSE SINGLE STRAND DNA
DAMAGE WHILE THIS MONTHLY
CULTURE CAN CAUSE DOUBLE
STRANDED DNA DAMAGE.
THIS INTERMEDIATE CAN LEAD TO
PROTEIN NI'M TRATION.
HAS BEEN SHOWN IN TERMS OF
ALCOHOLISM, CARDIOVASCULAR
AFFECT WHERE IT'S USED AS A
PRECONDITIONING AGENT AND WE
HAVE SHOWN THE EFFECT OF HNO IN
CANCER BIOLOGY AND INFLAMMATION
WHICH I WILL SHOW IN MY LATER
SLIDES.
SO UNDER PHYSIOLOGICAL
CONDITION, THESE ARE THE MOST
RELEVANT REACTIONS OF HNO.
AS I MENTIONED, IT REACT WITH
CERATYNE.
IT CAN ALSO REACT TO FORM METH,
HEMOGLOBIN SORT OF SPECIES.
IT CAN REACT TO FORM A GSSG .
THESE ARE SOME OF THE RELEVANT
RATE CONSTANTS AND THESE ARE NOT
ONLY IMPORTANT IN BUY OL JOE,
BUT THEY HAVE BEEN UTILIZED FOR
DEVELOPMENT OF METHOD FOR
DETECTION OF HNO.
SO ONE OF THE KEY REACTIONS
UNDER PIE LOGICAL COPY WAS
REACT, TO FORM RSNO WHICH CAN BE
CONVERTED TO PRODUCE HNO IN THE
PRESENCE OF GSH.
WHICH CAN FURTHER REACT WITH
THIOHS.
THIS SPECIES CAN LEAD TO
FORMATION OF GSSG SORT OF
SPECIES.
AS I MENTIONED, ONE OF THE
LIMITING FACTORS IS THE
DETECTION.
SO THESE ARE SOME OF THE METHODS
CHORE USED FOR DETENTION.
ANGELI'S SALT WHAT BEEN USED AS
AN HNO DONOR FOR A LONG TIME.
IT ALSO PRODUCES NITRITE.
WHICH IS SHOWN HERE IN THE RED.
IN THE PRESENCE OF HNO, IT
COMPLETELY CONVERTS TO MBNO TO
FORM SPEAKS AT 530 AND 56O
NANOMETER WHILE DNO DOES NOT
SHOW THESE PEAKS.
IT'S BASICALLY SAVED AS METHMYO
GLOBBEN.
IN THE PRESENCE OF GSH, WE DO
NOT SEE FORMATION OF THE
REDUCTIVE -- THE OTHER METHOD
WHICH IS COMMONLY USED IS THIS
ELECTRO CHEMICAL METHOD.
THIS IS AGAIN ELECTRODE FOR
NITRIC OXIDE DETENTION.
AS YOU CAN SEE, THE RED BARS
SHOW THE N.O. FORMATION AT
DIFFERENT PH IN PDS.
IN THE PRESENCE AT THE HIGHER
PH, WE SEE THAT THERE'S HNO
FORMATION WHICH IS HE HAVE DEN
FROM THE BLUE BARS.
THE DIFFERENCE SHOWS THE AMOUNT
OF HNO FORMATION.
AS YOU CAN SEE HERE THAT AS THE
PH OF THE -- AS THE PH OF THE
MEDIA INCREASES, AT THE LOWER
PH THERE IS NO HNO FORMATION.
RECENTLY THERE HAS BEEN A LOT OF
RESEARCH GOING ON AND THESE ARE
SOME OF THE PROBES THAT HAVE
BEEN GENERATED OVER THE YEARS.
SO THIS FLURRY HE -- FLUORESCENT
PROBE, SO THIS CAN DISTINGUISH
WITHOUT INTERFERENCE FROM N.O.
AND HIDE HID HIDE SPECIES.
BUT ONE OF THE DISADVANTAGES OF
THIS PROBE IS THIS CAN -- THIS
CAN BE REDUCED ALSO BY TILES.
SO WILL ACTUALLY SHOW
FLUORESCENCE WITH THE PROBE.
SO THERE HAS BEEN A LOT OF
RESEARCH GOING ON IN THE FIELD,
AND THIS COMPOUND WHICH WAS
RECENTLY PUBLISHED, THIS ONE
DOES NOT SHOW REACTIVE TILES AND
SHOWS MUCH MORE SPECIFIC --
SPECIFICITY FOR ETHANOL.
THESE FLUORESCENT PROBLEMS WHICH
WERE DEVELOPED IN WAKE FOREST
UNIVERSITY.
AND, AS YOU CAN SEE, HNO CAN
REACT WITH THE SPECIES WHICH
THEN -- WHICH THEN REACT AT THE
OX SITE AND THE FLURO IS
RELEASED.
ON REACTION WITH HNO IS FORMED
THIS SPECIES WHICH SHOWS A
STRONG FLUORESCENCE.
SO, AS I MENTIONED BEFORE, HNO
OF CHEMISTRY IS LIMITED BY
ITSELF A DIMERIZATION RATE, WE
NEED DONOR MOLECULES.
SO THESE ARE SOME OF THE DONOR
MOLECULES FOR N.O. WHICH ARE
COMMONLY USED AND THESE ARE SOME
OF THE DONOR MOLECULES FOR HNO.
SO THE MOST COMMON DONOR MONTHLY
-- MOLECULE HAS BEEN USED WHICH
IS USED FOR ANGINA PAIN.
THIS MONTHLY CULTURE WAS FIRST
DISCOVERED BY ALFRED NOBLE AND
THIS WAS BASICALLY THE PRECURSOR
FOR -- BUT WHEN WORKERS WERE
WORKING IN HIS FACTORY, HE
NOTICED THAT PEOPLE WOULD GET
HEADACHE BECAUSE OF EXPOSURE TO
VAPORS OF THESE MOLECULES.
THIS WAS BECAUSE THERE WAS
DILATION OF THEIR VEINS IN THEIR
HEADS WHICH WAS LEADING TO THIS
HEADACHE.
SO THIS HAS BEEN USED FOR A LONG
TIME FOR HEART RELATED ANGINA
PAIN.
THE OTHER MOLECULES, AND BOTH OF
THESE MOLECULES ARE -- SO TO
RELIEVE NITRIC OXIDE.
SO INVOLVED IS SETTLEMENT
EP-TWO, AND SEP-3 A 4.
THESE ARE MOSTLY USED DURING
HYPERTENSIVE.
HAS BEEN USED FOR INHIBITION OF
AGGREGATION.
SO WE ARE MOSTLY INTERESTED IN
MOLECULES BECAUSE THEY HAVE BEEN
USED UNDER SPONTANEOUS CONDITION
AND IT DOES NOT REQUIRE ANY
REACTION.
BUT LET ME GO OVER SOME OF THE
HNO DONORS FIRST.
THIS MONTHLY CULTURE AND THIS
MONTHLY CULTURE WAS SIN --
SINTHESIZED.
THEY ARE USED FOR STUDYING
MECHANICAL BIOLOGY OF HNO.
UNDER PHYSICAL CONDITION AND
SUFFICIENT THEN AN THEN IT
UNDERGOES A SMALL REACTION TO
TRANSFER THIS PROTON TO ON THE
NITROGEN.
WHILE SHOWS HNO PRODUCTION ONLY
AT A HIGHER PH.
SO THIS MOLECULE IS USED UNDER
CONDITIONS.
MAINLY FOR TREATMENT OF
ALCOHOLISM IS CYANAMIDE.
SO FOR TREATMENT OF ALCOHOLISM
EVEN THOUGH THIS IS APPROVED IN
JAPAN AND EUROPEAN COUNTRY, IN
U.S., IT'S NOT APPROVED.
THE OTHER HNO DOPER IS THIS
CLASS OF MOLECULES AND RECENTLY
THERE HAS BEEN COMPOUNDS WHICH
ARE ACTIVATED TO PRODUCE HNO.
SO WE LIKE TO WORK -- BECAUSE
THESE CAN BE EASILY BY DIFFERENT
SUBSTITUTIONS TO RELEASE NHO
VERSUS HNO.
SO WHEN -- IT'S A PURE HNO DONOR
AND IT -- PURE N.O DOPER IS A
PRIMARY AMINE.
AND THIS RATIO ACTUALLY DEPENDS
ON THE PH OF THE SOLUTION.
SO WE CAN CHANGE THE PH AND MAKE
IT REALLY COMPLETELY PURE --
VERSUS HNO.
THIS CLASS OF COMPOUND WAS
EXTENSIVELY USED AND DEVELOPED
BY DR. KIEFER IN NCI AS A RICH
TOOL AS WELL AS TO -- AS WELL AS
FOR DIFFERENT PATHOLOGICAL
PROCESSES.
SO ONE OF THE MAIN -- ONE OF THE
IMPORTANT THINGS ABOUT THESE
MOLECULES IS THEY CAN BE AT THE
LOCATION.
SO THESE ARE SOME OF THE
MOLECULES WHICH WERE DEVELOPED
IN DR. KIEFER'S GROUP, AND AS
YOU CAN SEE HERE, AS THE IMMUNE
SECOND CHANGES THE LIFE CHANGES.
IT HAS A HALF LIFE OF ONLY TWO
SECOND WHILE THE OTHER HAS A
HALF LIFE OF 20 HOURS.
THE HALF LIFE CAN INCREASE TO UP
TO SIX DAYS OR MORE.
THE PRIMARY CAN, AS I MENTIONED
RELIEVE BOTH NOR THIS PROTON
HERE CAN UNDERGO REACTION TO
ASSIST THIS AND THEN THIS
MONTHLY -- MOLECULE.
IT BASICALLY UNDERGOES THIS
PROCESS TO RELIEVE TWO MOLECULES
OF N.O.
THIS REACTION CANNOT FOR
SECONDARY.
250 MANO NANOMETER, THE REASON
WE BASICALLY USE THIS ABSORBENT
BAN IS BECAUSE THESE CAN HAVE
NIGHT RIGHT AND OTHER IMPURE
TEASE.
YOU MAY NOT GET THE RIGHT
CONCENTRATION.
SO WE WERE ALSO INTERESTED IN
EXPANDING THE LIBRARY OF PRIMARY
BASED SO WE COULD GET MORE HNO
DONATING COMPOUND.
AND THIS REFERS THE PSYCH LICK
PRIMARY AMINE SO WE MADE CPNO OR
CH AND CYCLO BASED AND AS YOU
CAN SEE HERE, THE HALF LIFE
CHANGES INTEREST LIKE 3.7 MINUTE
TO A MAXIMUM OF 6.4 MINUTE.
SO THE HALF LIFE DID NOT CHANGE
MUCH.
SO FOR THE SECOND KNOWN, THE
HALF LIFE DEVIATION WAS MUCH
MORE.
SO THEN WE LOOKED AT HOW MUCH
NOMH IS BEING RELEASED FROM
THESE MOLECULES AND WE FOUND
RELEASED THE MAXIMUM AMOUNT OF
HNO AT THE PHYSIOLOGICAL PH.
THESE MOLECULES WHICH DID NOT
SHOW THEMSELVES, AND AS
EXPECTED, THE ONLY WOULD BE --
SO AT THE LOW CONCENTRATION,
THEY ARE NOT BY THEMSELVES.
SO SINCE THE HALF LIFE DIDN'T
CHANGE MUCH, WE SYNTHESIZE BY
USING THIS ACOM.
SO UNDER IN THE PRESENCE, THIS
MOLECULE WHICH HAS A PEAK AT 240
NANOMETER SHIFT TO 250 WHICH IS
THE PEAK AND THEN IT DECOMPOSES
WITH THE HALF LIFE OF 5.8 MINUTE
WHICH IS BASICALLY THE SAME FOR
THE PARENT MOLECULES.
BUT IN THE PRESENT WHEN THERE IS
NO PRESENT, THIS 240 PEAK
DEPOSES WITH A CLEAN POINT.
SO IT IS NOT GOING THROUGH THE
PARENT MOLECULE.
SO THE FURTHER INVESTIGATED THE
MECHANISM OF DECOMPOSITION AND
WE FOUND THIS THIS PROTON IS
GOING TO BE EASILY AND THEN IT
CAN TPOPL THIS INTER MOLECULAR
RING WHICH CAN THEN FORM THESE
MIGHT TRICK WHICH DEKPOES TO
FORM HNO.
SO THEM WE HAVE THIS AUTO
PROTECTION AND THEN THERE IS NO
-- THESE MOLECULES ARE PURE HNO
DONOR.
SO WE QUANTIFIED THE AMOUNT OF
HNO PRODUCTION AND IT WAS 77%.
SO IN OTHER LABS, WE ARE MAINLY
INTERESTED IN LOOKING INTO THE
EFFECT OF VARIOUS HNO AND HNO
DONATING MOLECULES IN BREAST
CANCER.
SO THESE ARE SOME OF THE BREAST
CANCER STATISTICS AND I WOULD
NOT -- I WOULD SKIP TO IT.
I JUST WANT TO MENTION THAT WE
MAINLY WORK ON TREUL -L NEGATIVE
BREAST CANCER WHICH ARE ESTROGEN
RESIT ENTER NEGATIVE.
SO TRIPLE NEGATIVE BREAST CANCER
ACCOUNT FOR ONLY 15% OF BREAST
CANCER PATIENTS BUT THESE
PATIENTS HAVE A POOR SURVIVAL
RATE.
SO INFLAMMATION IS A KEY DRIVER
OF TUMOR PROGRESSION IN BREAST
CANCER ENOUGH TO ENZYMES ARE TWO
OF THE KEY ENZYMES INVOLVED IN
THIS PROCESS.
IN CANCER THE CHRONIC
INFLAMMATION CAN REVERT FROM
LIFESTYLE, OBESITY, INFECTION OR
AUTO IMMUNE DISEASE WHICH CAN
THEN LEAD TO INCREASED LEVEL OF
NITROGEN SPECIES, PRO
INFLAMMATORY PROTEIN
REARRANGEMENTS.
IN THE PHYSIOLOGICAL SYSTEM WE
HAVE IMMUNEO SURVEILLANCE.
MOST OF THE TIME THESE WHICH
ALER OUR IMMUNE SYSTEM IS ABLE
TO TAKE CARE OF IT.
UNDER SOME CIRCUMSTANCE, SOME
CIRCUMSTANCES, THIS CHRONIC
INFLAMMATION CONDITION CAN LEAD
TO FORMATION.
THESE ARE SOME OF THE COMMON
THERAPIES WHICH ARE USED FOR
BREAST CANCERS.
ONE OF THE COMMON THERAPIES IS
RADIATION, THEN WE HAVE HORMONE
THERAPY LIKE USE OF TAMOXIFEN,
THEN CHEMOTHERAPY IS USED AND
IMMUNEO THERAPY CAN ALSO BE USED
EVEN THOUGH IT IS NOT THIS
POPULAR IN BREAST CANCER.
BUT DRUG RESISTANCE IS ONE OF
THE KEY POB FOR TRIPLE NEGATIVE
BREAST CANCER PATIENTS.
SO OUR GOAL IS TO DEVELOP NEW
THERAPIES FOR THIS TYPE -- THE
TRIPLE NEGATIVE BREAST CANCER
PATIENT.
AS I MENTIONED, NOS-2 IS
INVOLVED IN PROGRESSION OF
VARIOUS CANCER, AND THE TYPE OF
CANCER WHICH ARE SHOWN HERE IN
GREEN SHOW POOR PROGNOSIS
RELATED TO NOS-2.
AND AS YOU CAN SEE, BREAST
CANCER IS ONE OF THEM.
IN CANCER BIOLOGY, MIGHT TRICK
OXIDE SHOWS A CONCENTRATION AS A
LOW CONCENTRATION.
IT CAN LEAD TO WHICH CAN LEAD TO
MUTATION IN THE DNA.
IT CAN ALSO LEAD TO INCREASEED
ANTI-GENESIS.
BUT AT THE HIGHER CONCENTRATION
OF MIGHT TRICK OXIDE, IT LEAD TO
INHEUBITION WHICH CAN LEAD TO
TUMOR CELL DEPOSITIONTHS.
SO IN OUR LAB WE USE AND WE HAVE
FOUND THAT DEPENDING ON THE TIME
AND FLUX OF NITRIC OXIDE IT CAN
LEAD TO ACTIVATION.
FOR EXAMPLE, AT THE LOW NITRIC
OXIDE THIS LEVEL BASICALLY CAN
BE CORRELATED TO THE AMOUNT
REDUCED BY NOT -- NOS THREE.
AT THE HIGHER CONSERVATIVE
TRAYING, B-53 IS FOSSILLATED AND
THEN THERE CAN BE SIGNALING SUCH
AS MODIFICATION OF RESIDUE.
SO THIS CONCENTRATION CAN BE
ACHIEVED BY NOS TWO.
SO BASED ON THE FLUX OF NITRIC
OXIDE IN THE CELLS, THE OUTCOME
OF THE DISEASE CAN BE MODIFIED.
SO FROM A THERAPEUTIC POINT OF
VIEW THERE CAN BE PRO
APPROACHES.
OR WE CAN INCREASE THE AMOUNT OF
NOS TWO.
SO ONE OF THE PREVIOUS IN
COLLABORATION WITH SHOWED IN A
PATIENT COHORT THAT 73% OF
BREAST CANCER PATIENT HAVE HIGH
NOS TWO EXPRESSION AND THEM
BASED ON THEIR -- FOUND THAT
ER-NEGATIVE PATIENT SHOW POORER
SURVIVAL.
SO 92% OF DECEASED ER-NEGATIVE
HAD HIGH NOS-2.
A RECENT PATIENT COHORT DONE IN
IRELAND IT IS FOUND 100% OF THE
DECEASED PATIENT HAD HIGH NOS 2.
SO THE OTHER ENZYME WHICH PLACES
INFORMATION IS COX-2.
THIS IS THE MECHANISM OF ACTION
SO CAN BE CONVERTED BY FOSSILIZE
ARE AN ACID WHICH IS THE
SUBSTRATE FOR COX ENZYMES WHICH
CAN LEAD TO FORMATION OF
DIFFERENT -- SO IN -- COX-2 HAS
PEN FOUND TO BE CORRELATED TO
DIFFERENT TYPES OF CANCER WHICH
ARE LISTED HERE AND BREAST
CANCER, AS WE CAN SEE HERE,
SHOWS OR LAYINGS WITH INCREASED
COX-2 LEVEL.
SO COX-2 -- EXIST IN TWO
DIFFERENT ICEO FORMS, COX 1 AND
COX-2.
THEN WE LOOK AT THE EXPRESSION
OF COX-2, WE FOUND THAT THE
PATIENT WHICH HAD HIGH COX-2 AND
HIGH NOS 2, THEY HAD A HAZARD
RATIO OF 20.
SO COMPARED TO THIS IS HIGH
NOS-2 ONLY, THE HAZARD RATIO
INCREASES FURTHER COMPARED TO
PATIENTS WITH ONLY HIGH NOS 2.
SO IN OUR LAB WE ARE INTERESTED
IN LOOKING AT THIS GROUP OF
PATIENTS, SO WE HAVE BEEN
STUDYING THE EFFECT OF THIS
GROUP BUT I AM NOT GOING TO GO
MUCH INTO THAT TODAY, BUT I'M
GOING TO TALK ABOUT SOME OF THE
MOLECULES WHICH WE HAVE
SYNTHESIZED IN OUR LAB.
THESE ARE MOLECULES -- TO
DIFFERENT ENDSTAGE -- DRUGS.
DRUGS WHICH WE HAVE USED WHICH
IS A REVERSIBLE INHIBITER OF COX
1 AND COX-2, AND WE HAVE ALSO
USED A REVERSIBLE INHIBITER OF
ONLY COX-2.
AND AS FOR THE DILATESES WE
COMPARED N.O. DONOR TO I PANEL
WHICH IS ONLY THE HNO DONOR.
SO HAVE BEEN ARE COMMONLY USED
TO ALLEVIATE PAIN AND
INFLAMMATION AND THESE ARE SOME
OF THE COMMON NSAID MOLECULES.
THIS IS RESPONSIBLE FOR
CONVERSION OF ACID -- SO ONE OF
THE MAIN SIDE-EFFECTS OF END
STAGE -- COX 1 AND COX-2, THIS
LEADS TO GASTRIC ULCERATION
RESPONSIBLE FOR MAINTENANCE OF
STOMACH LINING.
ANOTHER CLASS OF MOLECULES WHICH
WERE NOPE AS COX WHICH WERE
SPECIFIC COX-2 INHIBITORS WERE
SYNTHESIZED.
SINCE THESE MOLECULES ARE LARGER
IN SIZE, THEY FIT IN THE ACTIVE
SITE OF COX-2 WITHOUT COX 1.
SO EVEN THOUGH IT'S COX-2
SPECIFICALLY ON LONG-TERM USAGE,
IT'S LED TO CARDIOVASCULAR
INCREASE IN CARDIOVASCULAR
EVENTS.
SO FOR THIS REASON, WHICH WERE
TAKEN OUT OF THE MARKET AND
AMERICA -- MERCK HAD TO FACE A
HUGE LAWSUIT.
THE APPROACH OF N.O. DONORS AND
HNO DONOR BECAUSE AS I MENTIONED
EARLIER, N.O. CAN INCREASE, N.O.
CAN PREDICT THE STOMACH LINING
AS WELL.
WE THOUGHT THAT THESE CONJUGATED
MOLECULES CAN PREDICT, PROTECT
AGAINST THE SIDE-EFFECTS OF
ASPIRIN.
WE WANTED TO MAKE SURE SO WE
STUDIED AFFECTS OF THESE
MOLECULES.
AS WE CAN SEE HERE, THIS IS THE
CONTROL GROUP AND THIS IS THE
ASPIRIN GROUP.
WE BASICALLY GAVE THEM A HIGH
DOSE OF ASPIRIN AND THE QUESTION
LENT DRUG AND THEN THE STOMACH
LEADINGS WERE COUNTED AFTER SIX
HOURS.
AND ON TREATMENT OF ASPIRIN,
THERE WAS A HUGE INCREASE IN
ULCERATION INDEX.
THIS WAS THE FIRST TIME WE
SHOWED THAT ASPIRIN CAN PROTECT
AGAINST ULCER GENESIS.
THE OTHER SIDE-EFFECTS OF THIS
CLASS OF COX-2 CLASS OF
MOLECULES WAS HEART-RELATED
PROBLEMS.
SO WE STUDIED THE AFFECT OF
ASPIRIN ON CARDIOVASCULAR
MYOCYTES.
IT BASICALLY INCREASES -- IT'S
BASICALLY PRECONDITIONS THE
HEART AGAINST HEART ATTACKS SO
WHEN -- IN ANY EVENT OF A STROKE
THAT DAMAGE IS MUCH LESS.
WE ALSO WANTED TO MAKE SURE IT
REMAINS THE PROPERTY.
AS YOU CAN SEE, IT STILL
MAINTAINS THE PROPERTY COMPARED
TO ASPIRIN.
SO SIMILAR ANTI-INFLAMMATION
PROPERTIES.
INVESTIGATED THE EFFECT OF THESE
MOLECULES ON DIFFERENT BREAST
CANCER WHICH IS ER-NEGATIVE
BREAST CANCER AND WE USED MB-7
WHICH IS ANTI-POSITIVE BREAST
CANCER AND A NON-TUMOR ORIGIN.
AS YOU CAN SEE HERE, ASPIRIN
SHOWED THE MOST TOXICITY.
IT ALSO -- THE DATA WAS SIMILAR
IN BEFORE.
BUT DNO ASPIRIN MOLECULE BUT IT
WAS STILL PRE EXHAUSTED.
THE INTERESTING THING ABOUT
THE'S PRUP OR RELEASING ASPIRIN
IS IT DID NOT SHOW ANY TOXICITY
ON NON-TUMOR -- LOOKED INTO THE
EFFECTS OF THESE MOLECULES USING
A BRANCHING ASPECT AND THIS IS
THE CONTROL AND AS YOU CAN SEE
IT SHOWS A BRANCHING AND THIS
BRANCHING WAS INHIBITED BY LOW
DOSAGE DNO MOLECULE.
THEM WE STUDIED THE DNA DAMAGE,
AND SO THIS IS THE CONTROL, THIS
IS THE REPAUS -- POSIACTIVE
CONTROL.
SHOWED DNA DAMAGE BUT N.O.
RELEASING DN.O.'S PRUPB DID NOT
SHOW DNA DAMAGE.
THE OTHER THING WHICH WE
INVESTIGATED WAS THE ABILITY OF
THESE MOLECULES TO INHIBIT
COX-2.
SO USE -- AND CAME IN THIS
PURPOSE.
AND WE FOUND THAT ASPIRIN
INHIBITED LEVELS IN A
CONCENTRATION DEPENDENT WAY
WHILE DNO ASPIRIN DID NOT HAVE
MUCH EFFECT.
WHILE PART OF THE EFFECT IS FOR
SURE FROM THE ASPIRIN, WE HAVE
ALSO LIFTED THE AFFECT OF A PURE
HNO DONOR AND WE HAVE SHOWN THAT
IT CAN INHIBIT COX-2.
WE BELIEVE THE REASON FOR THIS
CAN BE SO THERE IS A VERY KEEN
ADAPTIVE SITE OF COX-2 WHICH CAN
UNDERGO REDUCTIVE IN THE
PRESENCE OF HNO OR IT MAY BE
DEPENDENT PROCESS.
SO WE HAVEN'T INVESTIGATED THE
MECHANISM BUT THESE ARE THE MOST
TWO COMMON PATHWAYS WHICH HAVE
BEEN SHOWN FOR DIFFERENT
DONATING MOLECULES.
WE ALSO INVESTIGATED THE EFFECT
OF HNO RELEASING ASPIRIN AND DNO
ASPIRIN BY GAPDHIN A
CONCENTRATION DEPENDENT MANNER.
WHILE DNO ASPIRIN DID NOT SHOW
ANY EFFECTS.
IT HAS BEEN SHOWN DUE TO THE
INHEUBITION OF THE RESIDUE AT
THE ACTIVE SITES.
AND THIS IS AN IRREVERSIBLE
NOTED FICTION.
THEN WE INVESTIGATED THE EFFECT
OF THESE MOLECULES ON OUR ANIMAL
POD -LZ AND WE USED NB 38 CELLS
LABELED WITH GFP.
THIS IS OF A CONTROLLED GROUP.
THE ANIMALS WERE INJECTED WITH
BMSO.
SIGNIFICANTLY REDUCED THE
PRIMARY TUMOR SITES.
WE MEASURED THE TUMOR SITE AS
WELL AND THIS IS THE QUANTIFIED
DATA FROM THE TPHRAOUR
FLUORESCENCE AND WE LIFTED THE
AFFECT OF THESE COMPOUND ON
METASTASIZED BY THE BRAIN BUT WE
DID NOT SEE A SIGNIFICANT
EFFECT.
SO THE OTHER ASPECT, AS I
MENTIONED, IS TO COMPLETELY
INHIBIT NOS 2.
STUDIED THE AFFECTS OF IMMUNEO
WHICH IS A KNOX 2 INHEUB TER ON
USING THE SAME MODEL, AND SHE
FOUND THAT IMMUNEO DECREASES THE
TUMOR VOLUME AS WELL.
SHE ALSO FOUND THAT IMMUNEO
SIGNIFICANTLY DECREASED THE
METASTASIZE TO THE BRAIN.
WE THOUGHT THIS WAS REALLY
INTERESTING BECAUSE WE DID NOT
SEE THAT WITH OUR DONEATING
MOLECULE.
RECENTLY MB ANDERSON HAS ALSO
RARIFIED THESE STUDIES AND SHE
ALSO FOUND SIMILAR RESULTS AND
THEY ARE CURRENTLY PLANNING TO
TAKE THESE CLINICAL TRIALS SO
THEY ARE GOING TO START A
CLINICAL TRIAL ON THESE.
SO APART FROM THE MOLECULES
DISCOVERED IN OUR GROUP, I'M
GOING TO GO OVER A FEW OTHER
MOLECULES WHICH HAVE BEEN
STUDIED BY DIFFERENT GROUPS.
SO THIS MOLECULE WAS AGAIN
DEVELOPED AT NCI AND
DR. KIEFER'S GROUP SHOWED
SIGNIFICANT DECREASE OF TUMOR
VOLUME IN DIFFERENT CANCER CELL
LINES.
ISSTUDIED IN BREAST CANCER,
MELANOMA AND IN ALL OF THEM
THOUGHT DECREASE IN TUMOR
VOLUME.
THIS MOLECULE RELEASES NITRIC
OXIDE IN THE PRESENCE OF GLUTTA
-- WHICH CLEVES TO CLEAVAGE OF
THIS BOND HERE AND THEN THIS
PART RELEASES TWO EQUAL LENGTHS
OF NITRIC OXIDE.
SOME OF THE OTHER MOLECULES
WHICH HAVE BEEN DEVELOPED ARE IN
IN NORTH CAROLINA.
HE USED THESE POLY -- BASED
MOLECULES AND SHOWED THIS THESE
CAN ACT AS ANTI-BACTERIAL
AGENTS.
ANOTHER GROUP SHOWED THAT WHEN A
STENT IS COATED -- DILATE, IT
DOES NOT FORM A SCAR TISSUE
AROUND IT AND DOES NOT BLOCK THE
STENT.
SO THESE WERE MUCH MORE
EFFECTIVE AND IT DID NOT GET
BLOCKED PAIRED TO THE CONTROL,
THE NONE QUOTED VERSION.
THEN RECENTLY ANOTHER GROUP
SYNTHESIZED THIS MOLECULE, SO
THIS IS A MACRO MOLECULE WHERE
IT'S LOADED WITH THEM OF CALCIUM
PHOSPHATE COATED SURFACE AND THE
SURFACE IS QUOTED BY HYDROGEN
AGAIN RATER.
THIS ACTIVATES PRODUCTION OF PRO
TONS WHICH REACTS WITH THIS
CALCIUM PHOSPHATE AND ON THIS
REACTION, CALCIUM PHOSPHATE WILL
FORM CALCIUM HIGH ANTHRAX ITE
WHICH WILL RESOLVE IN THE
CONDITION AND DILATES WILL BE
RELEASED.
THEN DECOMPOSES TO FORM NITRIC
OXIDES.
THEY USED -- THEY STUDIED THIS
MOLECULE LOOKING AT THE ONE
HEALING IN CORONA AND THEY FOUND
THAT COMPARED TO THE CONTROL
GROUP THESE QUOTED PARTICLES LED
TO AN INCREASE WOUND HEALING IN
A SHORT TIME AS SHORT OF TWO
DAYS.
SO I MENTIONED THIS ROUTINE
COMPLEX EARLIER, SO THIS STUDY
WAS DONE BY ONE OF OUR
COLLABORATOR IN COLLABORATION
WITH OUR GROUP.
AND THIS ROUTINE MOLECULE IS
ACTIVATED BY TILES TO RELEASE
HNO.
WE IN OUR LAB LOOKED AT THE
ANTIGENIC AND LOOKED AT
INHIBITION, N.O. HIP
STABILIZATION AND WHEN THIS
MOLECULE WAS ADDED, IT LEAD TO
HIP-1 ALPHA.
SO THE OTHER CLASS OF MOLECULE
WHICH HAS BEEN USED AS HNO
DOPERS, SO THIS IS ALSO A RECENT
STUDY IN AN GEO -- ANGIOTAM.
HNO RISKING MOLECULES IS ALREADY
IN THE CLINIC IN SOME PARTS OF
THE WORD.
AND ABUSED IN CLINIC AS AN
ALCOHOL DETERRENT.
USED FOR TREATMENT OF SICKLE
CELL AMEAN I CAN'T.
ALSO I WANTED TO POINT OUT THAT
THERE ARE MANY STARTUP COMPANIES
WHICH HAVE BEEN BUILT AROUND
RESEARCH AND I'M NOT JUST SAYING
THIS THESE MOLECULES ARE
IMPORTANT BECAUSE WE HAVE PATENT
ON IT, AND SO I CAME ACROSS THIS
COMPANY WHICH IS A REALLY NEW
COMPANY AND THEY WORK ON N.O.
DOPER BASED ON SOLID SUPPORT AND
THEY CAN CONTROL THE RATE FROM
FAST DONATING TO SLOW DONATING
MOLECULES AND THEY USED THESE
MOLECULES FOR DIFFERENT
MICROBIAL DISEASES AND THEY HAVE
MANY MOLECULES IN PHASE TWO AND
PHASE THREE TRIALS FOR TREATMENT
OF HERPES AND OTHER DISEASES.
THIS COMPANY WAS BASED ON HNO
RESEARCH, AND THIS -- THEY
STUDIED THE EFFECT OF HNO IN
HEART RELATED AILMENTS AND THIS
COMPANY WAS LAST YEAR BOUGHT BY
BRISTOL MYER AND THEY ARE
FURTHER DEVELOPING THESE
MOLECULES AND RIGHT NOW IT IS IN
THE PHASE THREE CLINICAL TRIAL.
AND BOTH OF THESE ARE
PHARMACEUTICAL COMPANY AND
DOJINDO I THINK.
THEY MAINLY WORK ON THE
BLUEPRINT OF RESEARCH
TECHNOLOGY, SO THEY ARE
DEVELOPING THE METHOD FOR
DETECTION OF ETHANOL COMPOUNDS
AND OTHER RESEARCH DEVICES FOR
STUDYING AND CHARACTERIZATION OF
DIFFERENT PROTEIN MODIFICATIONS
BY NONH.
SO IN CONCLUSION I WOULD LIKE TO
POINT OUT THAT DONOR COMPOUND
ARE REALLY IMPORTANT IN THIS
FIELD BECAUSE FOR BOTH N.O. AND
HNO BECAUSE NOT ONLY FROM
RESEARCH POINT OF VIEW BUT ALSO
FROM THERAPEUTIC POINT OF VIEW
BECAUSE WE WANT TO CONTROL THE
LEVELS OF THESE BIO MOLECULES
RELEASE, AS FOR N.O., THE MAIN
CHEMISTRY IS BASED ON MEDIATED
CHEMISTRY.
AND FOR N.O. IT IS MAINLY
RADICAL CHEMISTRY AND THESE LEAD
TO DIFFERENT PIE LOGICAL AFFECT
OF N.O. AND HNO.
AND AT THE END I WOULD LIKE TO
THANK MY ADVISERS DR. DAVID WINK
FORGIVING ME THIS COUNTRY AND
DR. TERRY MOODY FOR INVITING ME
TO GIVE THIS TALK.
I WOULD LIKE TO THANK LISA
RIDNOUR AND ROBERT CHENG.
HE HELPED ME WITH MANY OF THE
ANIMAL EXPERIMENT.
MADISON GREER IS A CURRENT POST
DOC STUDENT WORK WITH ME AND
VEENA SOMASUNDARAM WORKED ON
OTHER PROJECTS I DID NOT TALK
ABOUT TODAY.
THESE COMPOUND THAT I TALKED
ABOUT WERE SYNTHESIZED IN
UNIVERSITY OF ARIZONA DURING MY
PHD, AND DURING THIS TIME I WAS
A PRE DOCTORATE FELLOW AT NIH,
AND I DID THE BIOLOGICAL STUDIES
HERE.
AND I WOULD LIKE TO THANK ONE OF
HER COLLABORATORS SHARON WHO WAS
INSTRUMENTAL IN ANALYZING THE
PATIENT DATA.
AND THANK YOU ALL FOR YOUR TIME
AND THANK YOU FOR LISTENING AND
I'M OPEN SO MY QUESTIONS.
[ APPLAUSE ]
[INAUDIBLE]
>> YES.
>> SO IT'S REALLY A COMBINED
APPROACH OR CAN YOU -- ARE YOU
ABLE TO DETECT OUT WHICH IS
ACTUALLY ASYNERGISTIC EFFECT --
>> SO MINIMALLY WE WOULD QUENCH
THE HNO OR THE TILES AND SEE HOW
MUCH IT'S PRODUCING -- WHAT'S
THE RATIO OF N.O. AND HNO.
WE HAVE FOUND THE RATIO VARIES.
I SHOWED YOU MOLECULES SO THEN
IT'S PROTECTED, IT COMPLETELY
PRODUCES HNO.
SO IT IS A PURE HNO DOPER AT
THAT POINT EVEN THOUGH THAT HAS
A SHOCKER HALF LIFE.
BUT SOME OF THESE LIKE IPANEL
ASPIRIN HAS A MUCH LONGER HALF
LIFE.
WHEN WE DID THE EXPERIMENT, THEY
DID NOT FALL APART.
SO THEY ARE STILL PRODUCING HNO
IN THE MEDIA, PURE HNO.
IN THE PRESENCE OF THE SERUM, IT
WILL BE CLEAVED AND IT WILL
PRODUCE BOTH, SO IT REALLY
DEPENDS ON THE CONDITION.
IN THE CELL I'M GUESSING IF WE
DON'T REALLY HAVE -- WE DOPE
HAVE THE METHODOLOGY TO SAY HOW
MUCH PORTION OF LIKE N.O., WE DO
KNOW IT IS PRODUCING N.O.
BECAUSE WE HAVE LOOKED AND IT
SHOWS A MUCH HIGHER FLUORESCENCE
FOR HNO.
IT'S FIVE TIME AS HIGH AS HNO.
SO WE DO MOW IT IS PRODUCING HNO
IN THE CELL BUT WE DO NOT KNOW
HOW MUCH IS THE RATIO.
AND THE DECOMPOSITION PROCESS IS
BASED ON SPECIFIC -- SO WE
HAVEN'T LOOKED INTO THAT AS
INSPECT AS WELL.
[INAUDIBLE]
>> BUT I MEAN IT'S STILL REALLY
COOL BECAUSE FOR SOME REASON IT
IS NOT TOXIC TO NONE -- IT IS
NOT TOXIC.
IT IS NOT TOXIC TO CELLS.
LIKE BOTH DNO.
>> THANK YOU.
>> UM-HUM.
