SO IT'S A REAL PLEASURE TO INTRODUCE DR. AYATA, WHO IS HERE TODAY AS A CANDIDATE FOR THE CHIEF OF THE STROKE BRANCH. HE RECEIVED HIS M.D. AND Ph.D. IN TURKEY AND THEN DID HIS NEUROLOGY AT TUFTS AND THEN JOINED MASS GENERAL AS A STROKE FELLOW WHERE HE SAYS HE KNEW WALTER CORSHETTS VERY WELL. AND NOW IS CURRENTLY ASSOCIATE PROFESSOR IN THE DEPARTMENT OF NEUROLOGY AND RADIOLOGY. HE HAS DONE EXTENSIVE WORK WITH VARIOUS KINDS OF STROKE MODELS AND CURRENTLY HIS WORK REVOLVES AROUND SPREADING DEPRESSION AND STROKE AND OVERLAP WITH MYGRAIM. -- MIGRAINE. I HAD AN EXCELLENT CONVERSATION THIS MORNING ABOUT CLINICAL AND BASIC SCIENCE INTEREST. SO I'M EAGER TO HEAR HIS TALK. THANK YOU VERY MUCH. >> OKAY. THANK YOU VERY MUCH FOR THE KIND INTRODUCTION. AND INVITING ME TO COME AND SEE FIRSTHAND INTRAMURAL NIH PROGRAM IS ABOUT I HAVE TO ADMIT THAT WAS A COMPLETE UNKNOWN TO ME AND STILL IS AND LOOKING FOR THE REST OF THE DAY TO TALK WITH EACH OF YOU AND FIND OUT ALL ABOUT IT. IN PICKING THE TOPIC FOR TODAY, I HAD QUITE A FEW CHOICES. BUT I DECIDED TO GO WITH WHAT IS CLOSEST TO MY HEART, THAT IS DEPRESSION AND INJURY DELOCALIZATIONS. IF YOU CAN DIM THE LIGHT A LITTLE BIT. GREAT. SO I HAVE BEEN WORKING ON SPIN DEPRESSION FOR ALMOST 15 YEARS NOW AND PUBLISHED QUITE A FEW PAPERS AND I'M STILL AMAZED HOW UNDERAPPRECIATE AND UNDERRECOGNIZED THEY R. THEY HAVE CLINICAL TRANSLATIONAL RELEVANCE AND SHOWN IN HUMANS RECENTLY AS WELL WHICH I'M GOING TO SHOW PART OF THE DATA. AND SO I HOPE TO CHANGE THAT NOTION AND SEND YOU HOME A LITTLE BIT MORE THOUGHT ON SPIN DEPRESSION. SO LET ME STAWS, BEFORE I START I DON'T LIKE LEAVING PEOPLE BEHIND SO IF THERE'S ANYTHING YOU DON'T UNDERSTAND INTERRUPT AND WE ARE ASK, IT'S A SMALL GROUP AND WE CAN MAKE IT MORE INFORMAL. SO SPREADING DEPOLARIZATIONS ARE -- JUST GRAB ME WEAPON. ARE IN WORDS, SLOWLY PROPAGATING WAVES AT 2 TO 4-MILLIMETERS PER MINUTE, VERY SLOW PROPAGATION RATE, COMPLETE NEURAL AND GLIAL DEPOLARIZATION, AKIN TO SPREADING DEPRESSION. IF YOU KNOW WHAT SPIN DEPRESSION IS YOU KNOW WHAT INJURE DEPOLARIZATION IS BECAUSE THEY'RE IDENTICAL. THEY ORIGINATE FROM INJURED BRAIN, DOESN'T MATTER WHAT TYPE OF INJURY IT IS, COULD BE STROKE, INTRACELLULAR HEMORRHAGE, OR IT COULD BE A TRAUMA. AND ONCE IT ORIGINATE FROM THE NORMAL -- THE INJURED BRAIN TISSUE THEY PROPAGATE INTO NORMAL BRAIN TISSUE. QUITE A BIT. THEY SAY A PICTURE IS WORTS A THOUSAND WORDS, -- WORTH A THOUSAND WORDS AND I SAY IT'S WORTH A THOUSAND PICTURE, TECHNICALLY A THOUSAND PICTURES IN RAPID SEQUENCE. YOU GET THE IDEA. I HAVE A MOVIE OF BLOOD FLOW CHANGES DURING SOME OF THOSE INJURY DEPOLARIZATIONS IN A MOUSE STROKE MODEL. THIS IS A LATER SPECKLE METRIC MOVIE, IT'S AN OPTICAL IMAGING TECHNIQUE THROUGH IMPACT SKULL IN THE MOUSE BECAUSE IT'S THIN ENOUGH WE DON'T BREECH THE DOOR, WE JUST LOOK RIGHT THROUGH IT. HERE THE MODEL IS DISTAL MIDDLE FUSION. THE WAY WE DO IT IS TINY BURR HOLE OVER TO MCA AS IT GOES UNDER THE TEMPORAL BONE T SQUAMOUS PORTION OF TEMPORAL BONE AND WE PLACE A MICROVASCULAR CLIP AND JUST CLIP IT. ANDrq– THE RESULTING ISCHEMIA IS JUST ABOUT THIS. AND WHAT YOU WILL BE SEEING HOPEFULLY IN THE SCREEN IS PROPAGATING WAVE, I'LL POINT AS THEY COME. EVERYTHING IS COLOR QOADED THE MORE BLUE -- CODED. THE MORE BLUE THE MORE SEVERE ISCHEMIA SSM THIS IS A BLOOD FLOW MAP, FOUR MINUTES AFTER THE MCA ISK COLLUDED SO THIS DOESN'T -- IS OCCLUDED SO IT DOESN'T CAPTURE THE OCCLUSION POINT. HERE IS WHAT HAPPENS. HOPEFULLY AS SOON AS IT STARTS. HERE IS -- THIS WAS THE FIRST WAVE. HERE IS ANOTHER ONE. THIS IS FAST FORWARDED SO ALREADY 12 MINUTES, RIGHT? HERE IS ANOTHER ONE, A BIG ONE. MORE H COME. HERE IS -- MORE WILL COME. HERE IS ANOTHER ONE SPREAD BACK THIS WAY. THESE ARE EXAMPLES OF INJURY DEPOLARIZATIONSCH THESE FAST FORWARD THEY PROPAGATE 3-MILLIMETERS PER MINUTE. THIS IS SLOW PROPAGATION. HERE SAN EXAMPLE FROM THE RAT, THIS MODEL AND THIS IS AN EXAMPLE OF A CIRCLING INJURY DEPOLARIZATION. YOU WILL SEE THE DEPOLARIZATION THIS WAY. S THAT RAISER SPECKLED BLOOD FLOW MOVIE. AGAIN THIS IS A FAST FORWARDED MOVIE, FASTER THAN THE REAL TIME. , THIS IS ANOTHER INJURY DEPOLARIZATION FOR YOU. IT DOES HAVE IN ENCEPHLOPPIC BRAIN AS WELL. THIS IS A CAT OCCLUSION MODEL FROM TONY STRONG IN LONDON. THERE IS A CRANEIOTMY HERE,IOTTHROUGH IMPACT SKULL AND THESE ARE THROUGH ELECTRODES HE USED TO RECORD DEPOLARIZATIONS. ISCHEMIA IS INDUCED HERE BY CLIPPING -- THIS IS THE CORE AND PRETTY MUCH THE P NUMBER AND THIS IS NON-ISCHEMIC CORTEX. SO HERE IS WHAT HAPPENS IN THE CAT. >> THE IMAGING BLOOD FLOW YOU'RE SAYING (INAUDIBLE)? >> YES, BECAUSE THESE ELECTRODES AT THE SAME TIME THIS IS ON AND OFF, AT THE SAME TIME RECORD DEPOLARIZATION AS WELL SO AS BLOOD FLOW CHANGE IS HAPPENING YOU HAVE -- O I JOUST DON'T HAVE THOSE RECORDINGS HERE TO SHOW TO YOU. (OFF MIC) >> WE KNOW. WE HAVE DONE THOUSANDS OF RECORDINGS, YOU HAVE TO TRUST ME FOR THAT. I'LL SHOW YOU EXAMPLES OF THAT. THESE BLOOD FLOW CHANGES IS A SURROGATE TO GIVE YOU AN IDEA OF THE SPATIAL TEMPORAL PATTERN OF DEPOLARIZATIONSCH I'LL GET INTO THAT IN A SECOND. IT'S TRUE. IT'S A FAIR QUESTION. HERE IS ANOTHER LOOK AT IT, HERE IS THE WAVE GOING,TWO MERGING AND THERE'S ANOTHER EXTENSION OF IT UP INTO THE NON-ISCHEMIC BRAIN TISSUE. SO NOW, WHEN INJURE SPIN DEPRESSION HAPPENS IN OTHERWISE NORMAL BRAIN, IF YOU TRIGGER IT YOURSELF OR WHEN IT HAPPENS IN CASE OF MIGRAINE AURA, INJURY DEPOLARIZATION AN SPIN DEPRESSION ARE INDISTINGUISHABLE AS I SAY. THEY'RE IDENTICAL. WHEN THEY HAPPEN IN INJURED BRAIN THEY CAN HAPPEN IN CASE OF ISCHEMIA HEMORRHAGE OR TRAUMA. AND EVEN THOUGH SPIN DEPRESSION WAS KNOWN SINCE 1940s FROM (INDISCERNIBLE) WORK, WHO TERMED THIS DEPRESSION GAVE THE NAME TO IT, WE DID NOT KNOW THEY WERE HAPPENING IN THE PERIINJURED AREA UNTIL ABOUT 1986 WHEN IT WAS FIRST RECORDED. IT'S BEEN QUITE A FEW YEARS SINCE THEN, SPORADIC STUDIES CAME OUT BUT ONLY LATELY WE APPRECIATE THEIR CLINICAL IMPORTANCE, I'LL SHOW YOU IN THE NEXT SEVERAL SLIDES. FOR THOSE WHO DON'T KNOW WHAT SPIN DEPRESSION S I HAVE A SEQUENCE OF CARTOONS HERE TO GIVE YOU A BASIC IDEA WHAT SPIN DEPRESSION IS. AS YOU KNOW THE IAMS IN NORMAL BRAIN TISSUE ARE UNEVENLY DISTRIBUTED IN CELL MEMBRANES. EXTRA CELLULAR POTASSIUM IS LOW COMPARED TO INTRACELLULAR POTASSIUM, AN ORDER OF MAGNITUDE OF POTASSIUM GRAITD GRADIANT. INTRACELLULAR CALCIUM IS ALSO EXCEEDING LOW AND SODIUM IS DISTRIBUTED IN THE OPPOSITE DIRECTION, ENDOCELLULAR IS LOWER THAN EXTRA CELLULAR SODIUM. WHAT HAPPENS WHEN SUFFICIENTLY INTENSE STIMULUS SIMULTANEOUSLY DEPOLARIZES AND MINIMUM CRITICAL WORDING OF BRAIN TISSUE, AND THIS IS ESTIMATED TO BE 1 MILLIMETER CUBE IN RODENT BRAIN. THESE CELLS UNDERGO A SEQUENCE OF CHANGES THEIR MEMBRANE OPENING OF NON-SELECTED AT ION CHANNELS THAT DUMP THEIR MASSIVE INTRACELLULAR POTASSIUM BY MORE THAN TENFOLD, 40 TO 80 MILLIMOLAR CONSENT TREATION AND THIS IS HAPPENED THROUGHOUT THE TISSUES FLOODED BY POTASSIUM. AT THE SAME TIME THERE'S MASSIVE CALCIUM INFLUX THAT TRIGGERS UNCONTROLLED GLUTAMATE RELEASE. THERE'S EXTRA CELLULAR GLUTAMATE INCREASE, AGAIN, BAITING THE ENTIRE TISSUE -- BATHING THE TISSUE IN GLUTAMATE. AT THE SAME TIME THERE'S MASSIVE SODIUM INFLUX WHICH BRINGS WATER WIT INTO THE CELL AND CELLS SWELL. NOW, IT IS BELIEVED THAT THIS INCREASE EXTRA CELLULAR POTASSIUM IN GLUTAMATE DIFFUSES OUT TO NEIGHBORING NON-DEPOLARIZED CELLS AND TRIGGERED THE SAME DEPOLARIZATION CYCLE INTO CELLS SO THAT THIS SPREADING -- THIS FOCAL DEPOLARIZATION BECOMES A SPINNING DEPOLARIZATION BY WAY OF CONTY GIEWTY. SO THIS IS NOT A STAND SYNAPTIC CONNECTION THAT PROPAGATES THE DEPOLARIZATION, THIS IS PURELY CHEMICAL. IT DEFOR LA RISES THE SURROUNDING CELLS AND THAT'S WHY IT PROPAGATES SO SLOWLY, IT DEPENDS ON IONS AN RELEASE RATE. IF THE TISSUE IS NORMAL, NOT HYPOXIC OR ISCHEMIC THE CYCLE REVERSES ITSELF WHEN LESS THAN A MINUTE. THROUGH UPDATE BY ASTROCYTES, NEURONS AND VASCULATURE AND IT COSTS ATP, OXYGEN AND GLUCOSE, THE CONSUMPTION GOES UP. NOW ALL THESE MASSIVE IONIC SHIFTS CAUSE A CHARACTERISTIC NEGATIVE POTENTIAL SHIFT WHEN YOU RECORD EXTRA CELLULARLY USING ELECTRODES IN THE BRAIN. HERE IS AN EXAMPLE, THOSE PROPAGATING WAVES IF YOU HAVE AN ELECTRODE IN THERE WHICH WE HAD IN THE PAST, TO RECORD ELECTROPHYSIOLOGICAL CHANGES THIS IS WHAT YOU SEE. THERE'S A MASSIVE SHIFT, BECAUSE IT'S -- WHEN WE TALK EEGs, HERE YOU'RE RECORDING MILIVOLTS SHIFT AND THE EEG IS DEPRESSED. WHEN (INDISCERNIBLE) IN 1944 NAMED IT SPIPPING DEPRESSION BECAUSE HE WAS USING ONLY THE EEG TO DEFINE, TO DECK THE WAVES. WHAT HE SAW WAS DEPRESSION OF EEG ACTIVITY. SO HE CALLED IT SPREADING DEPRESSION WAVE. BUT IT'S KIND OF A MISNOMER BECAUSE THIS IS NOT A DEPRESSION IN THE SENSE THAT WE UNDERSTAND IT, WHEN WE TALK ABOUT DEPRESSION OF ACTIVITY WE TALK ABOUT HYPERPOLARIZATION, SUB PRESENTATION, QUIET TISSUE BUT THIS IS A SUPER DEPOLARIZATION. THE MEMBRANE POTENTIAL IS ZERO WE CANNOT TRIGGER AN ACTION POTENTIAL SO THERE IS NONE, THERE'S NO SYNAPTIC TRANSMISSION THAT'S WHY THE EEG IS FLAT, NOT FLAT BUT QUIET. IF YOU HAVE TWO ELECTRODES PLACED IN SERIESS FROM THE ORIGIN OF SPREAD DEPRESSION YOU CAN CONNECT THE SAME WAVE WITH A LAY TAN SI AND SOMETIMES BIGGER, THAT GOES TO SHOW YOU THAT IT'S A REGENERATIVE PROCESS, THIS IS NOT JUST A DIFFICULT WHAT HAPPENED THERE, THIS IS A REGENERATING PROCESS THAT REACHES WITHOUT BEING DIMINISHED TO THE SECOND OR REMOTE SITE. THIS TABLE SHOWS THE CONTRAST BETWEEN I DON'T KNOW IF YOU CAN SEE IT, THE PHYSIOLOGICAL DEPOLARIZATION THAT WE TALK ABOUT, THE ACTION POTENTIAL IN THE BRAIN, AND SPIN DEPRESSION. THE ACTION POTENTIAL TRAVELS AT MILLILITERS PER SECOND, THAT'S HOW WE MEASURE IT. MILLIMETERS PER MINUTE, IT'S 100,000-VOLTS SLOWER THAN PHYSIOLOGICAL NEURAL ACTIVITY. ITS AMPLITUDE IS MICROVOLTS IN ACTION POTENTIAL AND MILIVOLTS IN CASE OF SPIN DEPRESSION AN DURATION OF ACTION POTENTIAL IS MEASURED IN MILISECONDS BUT SPIN DEPRESSION LASTS UP TO A MINUTE. THAT'S A VERY DIFFERENT BREED OF DEPOLARIZATION YOU'RE TALKING ABOUT HERE. THAT'S WHAT SPREADING DEPRESSION IS AND INJURY DEPOLARIZATION ARE SPIN DEPRESSION WAVES THAT HAPPEN THAT ORIGINATE FROM INJURED BRAIN TISSUE. HERE IS AN EXAMPLE, TWO PLACES, I HAVE ONLY ONE TRACING FROM ONE OF THEM, THAT SHOW REPETITIVE WAVES OF SPIN DEPRESSION IN THE SETTING OF MOUSE FOCAL ISCHEMIA. THE GRAPH AT THE BOTTOM SHOWS TEN INDIVIDUAL EMPERIMENT WHERE IS THESE RECORDINGS WERE DONE TO SHOW YOU THE DISTRIBUTION OF THESE DEPOLAR SITION WAVES. EACH HORIZONTAL LINE INDICATE IT IS ONSET OF RECORDINGS AND THE END OF RECORDINGS DURING A 2 TO 3 HOUR PERIOD. AND EACH CIRCLE INDICATES ONE INJURE DEPOLARIZATION. SO YOU CAN SEE THAT THEY KEEP HAPPENING THROUGHOUT THE FIRST THREE HOURS AND I'LL SHOW YOU LATER THAT THEY KEEP ON COMING AFTERWARDS AS WELL. THEY'RE RELATIVELY RANDOM, IN THIS SPACE BUT THEY HAPPEN IN EVERY SINGLE ANIMAL, THERE'S NOT A SINGLE ANIMAL THAT DOESN'T HAVE A INJURY DEPOLARIZATION. IN FACT THEY KEEP HAPPENING UP TO FOR UP TO 24 HOURS, AT LEAST IN THE RAT. A NUMBER OF YEARS BACK JED HARTINGS HAS DONE AWAKE RAT RECORDINGS OF FILAMENT MCO AND DETECTED THE OCCURRENCE OF INJURY DEPOLARIZATIONS IN STROKE FOR UP TO 24 HOURS. THE PATTERN WAS INTERESTING, IN PERMANENT MCO THEY KEPT COMING AND THERE WAS A SLIGHT HINT OF SECOND PHASE AROUND 12 HOURS, WHAT WAS INTERESTING IS EVEN IF YOU REPER FUSED AFTER TWO HOURS, THIS IS A TWO HOURS TRANSIENT MCO, IF YOU REPER FUSE HAT THIS POINT THOUGH IT STOPPED A BIT THEY STARTED COMING BACK UP AGAIN. THERE'S NO ISCHEMIA HERE. INJURED TISSUE IRREVERSIBLY INJURED TISSUE CAPABLE OF TRIGGERING DEPOLARIZATION THOUGH FLOOD OF BLOOD IS BACK TO NORMAL SO YES, THEY DO HAPPEN UP TO 24 HOURS. A VERY IMPORTANT CONCEPT TO DIFFERENTIATE SPIN DEPRESSION FROM INJURY DEPOLARIZATION OR PERIIMPOLARRIZATIONS, THE FACT THEY'RE PROLONGED QUITE OFTEN, AND THEY MAY BECOME PERMANENT. LET ME EXPLAIN WHAT THIS MEANS. HERE I HAVE A MOUSE FOCAL ISCHEMIA, I HAVE AN ELECTRODE IN THE CORE AND THE VERTICAL LINE THERE IF YOU CAN SEE THE LIGHT FAINT LINE INDICATES THE TIME OF ON TIEWTION. WITHIN A COUPLE OF MINUTES, IN THE CENTER OF THE ISCHEMIC TERRITORY, IF YOU HAVE AN ELECTRODE THIS IS WHAT YOU RECORD. IN A COUPLE OF MINUTE IT IS TISSUE UNDERGOES WHAT WE CALL OXIDATIVE DEPOLARIZATION. THIS IS THE SAME DEFOR LAIZATION THAT THE TISSUE UNDERGOES DURING SPIN DEPRESSION. BUT IT'S PERMANENT. WHEN I SAY PERMANENT, IT'S LONG LASTING AN GOES ON, NEVER RECOVERS AS LONG AS THE OCCLUSION IS THERE. IF YOU REMOVE IT AFTER 30 MIGHT BEES FOR EXAMPLE, THIS MAY REVERSE ITSELF. THAT DOESN'T MEAN TISSUE WILL SURVIVE, JUST A RETAIN THE ABILITY TO REGAIN MEMBRANE POTENTIAL. BUT THIS IS OXIDATIVE POLARIZATION, THIS IS IN THE CORE. IF YOU PUT AN ELECTRODE OUTSIDE ISCHEMIC TERRITORY YOU SEE REPETITIVE INJURY DEPOLARIZATIONS TRIGGERED FROM THE INJURED BRAIN. BUT IF YOU PUT AN ELECTRODE RIGHT AT THE BORDER ZONE BETWEEN CORE AND P NUMBER WHAT YOU SEE IS PROLONGED WAVES OF DEPOLARIZATION. THESE ARE PERIDEPOLARIZATIONS MUCH LONGER THAN WHAT YOU SEE IN NORMAL BRAIN BUT IT DOESN'T -- IT'S NOT PERMANENT, IT RECOVERS AFTER A WHILE. AND THE TIME SCALE HERE IS I BELIEVE THIS IS ABOUT FIVE MINUTES. I FORGOT TO PUT THIS ON. AND WHAT HAPPENS IS YOU HAVE REPETITIVE WAVES LIKE THIS, THAT CAN BECOME PERMANENT. WHERE THIS ELECTRODE WAS AT THIS TIME POINT ENDED UP BEING INCORPORATED INTO THE CORE THAT IS THE PERMANENT DEPOLARIZED BRAIN TISSUE AND NEVER RECOVERED FROM IT THAT'S AN EXPANSION OF THE CORE FOR YOU. HERE IS ANOTHER EXAMPLE, THESE ARE SPIN DEPRESSIONS TRIGGERED BY TOPICAL CHLORIDE APPLICATION CONTINUOUSLY AND REPETITIVE AND THEY'RE CRISP AND NARROW RECOVERING SPIN DEPRESSIONS. HERE ARE EXAMPLES AT THE SAME TIME SCALE OF INJURE DEPOLARIZATION IN CASE OF ANTI-OCCLUSION. THEY ARE PROLONGED AND SOMETIMES TOO PROLONGED, THIS IS MORE THAN TEN MINUTES, THEN RECOVERING AND FAILED TO RECOVER. MOST IMPORTANT ABOUT TEN YEARS AGO A GROUP OF AMERICAN AN EUROPEAN INVESTIGATORS CAME TOGETHER AND FORMED A STUDY GROUP, THEY TERMED COLLABORATIVE STUDY OF BRAIN INJURY DEPOLARIZATIONS, THEY CALL THEMSELVES (INDISCERNIBLE) THEY ACCOMPLISHED A GREAT FEAT BECAUSE THEY IMPLANTED THROUGHOUT MANY CENTERS SUB DO YOU RECALL ELECTRODE STRIPS IN PATIENTS WHO ARE UNDERGOING CRANEECTOMIES FOR ANY INDICATIONS FOR BRAIN INJURY. IT COULD BE STROKE, HELP CRANEECTOMY, IT COULD BE HEMORRHAGE OR HEAD TRAUMA. HERE IS AN EXAMPLE FOR MALIGNANT MCH STROKE AND HERE IS THE STRIP ON A CT. THE STRIP HAS SIX LEADS ON IT AND THESE ARE GOING TO GO INTO BIPOLAR RECORDINGS A VERSUS B AND B VERSUS SO FORTH. AND IN THIS EXAMPLE THE STRIP IS PLACED AT THE BORDER ZONE AND THEY TRY TO DO THAT AS MUCH AS THEY CAN. WHAT THEY HAVE ACCOMPLISHED TO SHOW WAS EVENTS JUST LIKE THIS. THESE ARE SURFACE ELECTRODES, NOT GO OFF MICROPIPETTES SO THE ELECTROPHYSIOLOGICAL SHAPE IS VERY DIFFERENCE, A LITTLE MORE PROLONG BECAUSE OF AVERAGING AT THE SURFACE. THEY ALSO SMALLER IN AMPLITUDE BUT THESE PROPAGATE FROM ONE LEAD TO THE OTHER. I'M SORRY ABOUT THIS SMALL STREEN, BUT WHAT YOU'RE NOT SEEING BUT IT'S THERE IS LINES SHOWING DIRECTIONALITY OF SPREAD, STARTS FROM CHANNEL D WHICH IS THE BACK CHANNEL CLOSER TO THE BORDER ZONE AND PROPAGATES TOWARDS A. MULTIPLE WAVES OF EEG DEPRESSION ACCOMPANIED BY DC FLUCTUATIONS SIMILAR TO SPIN DEPOLARIZATIONS. SO THIS NOW HAS BEEN SHOWN IN MANY DIFFERENT STUDIES ACROSS THE BOARD FOR DIFFERENT TYPES OF INJURIES, AND THERE'S VERY LITTLE DOUBT THESE ARE INJURY DEPOLARIZATIONS MUCH LIKE RECORDING ANIMALS. SO THEY HAPPEN IN MICE, RATS, CATS AND FINALLY IN HUMANS. BUT ARE THEY IMPORTANT? WHY DO WE CARE ABOUT THEM? VERY IMPORTANT, THEY IMPACT TISSUE OUTCOME IN A NEGATIVE WAY. THERE ARE SEVERAL WAYS OF DOING THIS. FIRST OF ALL, AS IrxD„ SAID AT THE BEGINNING SPINNING DEPRESSION REQUIRES ENERGY TO CROAFER, BECAUSE ITS MASSIVE IONIC SHIFT AND SOMETHING HAS TO CORRECT IT, SOMETHING HAS TO REVERSE OR RESTORE THE TRANSMEMBRANE GRADIANTS. THAT COSTS ATP AND CONSECUTIVETIVELY OXYGEN AND GLUCOSE. SO OXYGEN AND GLUCOSE CONSUMPTION GOES UP DURING INJURY DEPOLARIZATION AND THAT INCREASES MISMATCH. HERE SAN EXAMPLE, HERE I HAVE A REFLECTIVE IMAGING OF (INDISCERNIBLE) CONCENTRATION IN THE CORTEX. AGAIN, IMAGED THROUGH INTAK SKULL IN A MOUSE. THE SAME PROFUSION MODEL I SHOWED YOU EARLIER. THIS IS COLOR CODED. AND GREEN IS BASELINE AS YOU CAN SEE HERE, THIS IS NOW BEFORE THE MC OCCLUSION, WHAT THE BASELINE IMAGE LOOKS LIKE. AND THIS IS THE MOMENT OF MC OCCLUSION, THIS IS THE INITIAL DEPOLARIZATION AS DESCRIBED. IF YOU'LL LOOK CLOSELY HOPEFULLY THE COLOR CONTRAST WILL SHOW THAT. HERE IS THE FIRST WAVE OF INJURY DEPOLARIZATION. THIS ISOXI MOBILE CONCENTRATION GOING DOWN, BLUE INDICATES REDUCED CONCENTRATION. HERE IS A SECOND WAVE GOING DOWN. LOOK HOW BLUE THE TISSUE BECOMESCH THIS IS A SIGN OF TISSUE HYPOXIA, OXYGEN CONSUMPTION GOES UP DURING THESE WAVES. HERE IS ANOTHER EXAMPLE, THE COLOR MAP IS VERY DIFFERENT HERE BUTBLY BLU WILL BE A REDUCTION IN CONCENTRATION. THIS IS AGAIN (INDISCERNIBLE) IMAGED THE SAME WAY THROUGH INTAK SKULL BUT ORIENTATION IS DIFFERENCE. NOW THE HEAD IS TURNED UP, THERE IS THE CLIP THIS IS THE LATERAL EDGE OF THE HEMISPHERE, THIS IS MED LINE. (OFF MIC) THAT'S HARD TIME. AT THE SAME TIME THERE'S BAY ZILLION CONSTRICTION SO BY LOOKING AT THESE YOU CANNOT DECIDE THIS IS BECAUSE OF INCREASED OXYGEN CONSUMPTION BUT THE NEXT SLIDE AFTER THIS I BELIEVE SHOWS THE ELECTRODE RECORDINGS FROM THE BRAIN OF PO-2, NOT INTRAVASCULAR PO-2, THAT SHOWS A DIP IN OXYGEN TISSUE OXYGENATION. THAT DIRECT EVIDENCE THAT IS THE TISSUE PO-2 GOING DOWN AND TISSUE PO-2 GOING DOWN COULD BE A VASCULAR REFLECTION, THAT'S RIGHT. AND WHEN YOU LOOK AT -- (OFF MIC) >> YES. THERE ARE OTHER FACTORS TOO. I DON'T WANT TO GET INTO THIS BUT THERE ARE OTHER FACTORS TOO. THERE'S A PROBLEM WITH OXYGEN DID I HAVEIVITY. THE DIFFUSION DISTANCE OF THE CONSTANT OF OX GENERAL CHANGES. EVEN IF YOU HAVE INTRAVASCULAR OXYGEN AVAILABLE IT MAY NOT GET TO THE CELLS AS WELL AS NORMALLY, IT MAYBE CONSUMED MUCH FASTER. SO THERE ARE MANY OTHER ISSUES. BUT I WAS GOING TO SAY, OH, YEAH, NADH IMAGING. IF YOU LOOK AT THAT YOU SEE INCREASE IN ADH. AN INCREASE IN ADH BECOMES BECAUSE OF HYPOXIA. SO IT'S A ME TAB O OXIDATIVE PHOSPHORYLATION IF OXIDATIVE PHOSPHORYLATION SLOWS DOWN THERE'S BILL UP BECAUSE IT CAN'T FEED INTO THE MITOCHONDRIA AND GET RELEASED, GET OXIDIZED. SO THAT HAS ALSO -- THERE IS ANOTHER PIECE OF EVIDENCE THAT SUGGESTIONS THAT TISSUE HYPOXIA DOES HAPPEN. BECAUSE OF INCREASED CONSUMPTION BECAUSE NADH DOESN'T NECESSARILY GO UP DRAMATICALLY BY 20 FOLD IF YOU DIDN'T HAVE INCREASED DEMAND. WE ALSO KNOW THERE'S INCREASED GLUE CONSUMPTION SO -- GLUCOSE CONSUMPTION SO HOPEFULLY BY THE NEXT FEW SLIDES YOU'LL BE CONVINCED. ANOTHER EXAMPLE, THIS IS ANOTHER INJURY DEPOLARIZATION. LOOK HOW BLUE THE TISSUE GETS. THIS CAN BE IN PART BECAUSE OF THE BASAL CONSTRICTION. LARDLESS THE TISSUE IS HYPOXIC, VASCULARLY OR METABOLICALLY, AT THE END OF THE DAY IT DOESN'T MATTER. SO HERE IS THE PO-2 RECORDINGS CALLED HUMANS, BECAUSE WE MANAGE TO INCORPORATE A PO-2 ELECTRODE INTO THE SUB DO YOU RECALL STRIP THAT'S RECORDED IN REAL TIME IN ASSOCIATION WITH THE EEG CHANGES, THE TISSUE PO-2 AND THESE ARE INDIVIDUAL INJURY DEPOLARIZATION COMPRESSED TIME SCALE SO THERE'S A DIP IN THE SUBSEQUENT INCREASE AND THIS IS AN EXPANDED SCALE, KIND OF HARD TO SEE I SUPPOSE BUT THERE'S A DIP OF PO-2 HERE FROM BASELINE OF 35-MILLIMETER, ABOUT 15-MILLIMETERS THAT WAS ABOUT A MINUTE OR MORE. SO THESE INJURY DEPOLARIZATIONS ARE ASSOCIATED WITH TISSUE HYPOXIA. AS I SAID, GLUCOSE CONSUMPTION ALSO GOES UP DURING DEFOR LAIZATIONS. IN ANOTHER STUDY A MICRODIALYSIS PROBE WAS INCORPORATED TOGETHER WITH ELECTRODE STRIP AND WHAT THAT SHOWED IS THIS IS THE IMPORTANT PORTION OF THE GRAPH, THIS SHOWS THE INJURY DEPOLARIZATION AS DECKED BY ELECTRODE BUT AT THE SAME TIME WITH THE INJURED DEPOLARIZATION THERE'S INCREASE IN DIALYSIS LACTATE AND DECREASE IN GLUCOSE. THAT'S ONE EMPERIMENT, ONE EXAMPLE, THIS IS REAL TIME ON LINE MICROBIOME. HERE IS A SUMMARY OF SEVERAL PATIENTS. UPPER GRAPH IS LACTATE, LOWER IS GLUCOSE. THE DIP IN GLUCOSE DURING DEFOR LAIZATION IS VARIABLE FROM ONE PATIENT TO ANOTHER BUT IT'S ALWAYS IN THE DECREASED DIRECTION SO INCREASE OF GLUCOSE CONSUMPTION, IN ASSOCIATION WITH INCREASED LACTATE PRODUCTION DURING INJURY DEPOLARIZATIONS. SO I HOPE THIS IS YOUR COMMENT ABOUT THIS. THERE'S INCREASED METABOLIC LOAD ON THE TISSUE WHICH INCREASES. WHAT WAS MORE SURPRISING THOUGH, SPATIALLY. DISTANCE WISE, NO. IN HUMANS, IT DOESN'T GO -- HUMAN BRAIN IS RELATIVELY RESISTANT FOR INJURY DEPOLARIZATION. PEOPLE PROPOSE GLIAL OR ASTROSITIC DENSITY OR HIGH GLIAL NEURON RATE AND THERE MAYBE MORE THAN ONE CODE FOR THAT BUT WE DONE KNOW REALLY N. A MOUSE TRIGGERED HERE YOU CAN GO ALL THE WAY THROUGHOUT THE HEMISPHERE THROUGH HUMAN IT'S LESS THAN A (INAUDIBLE). WE NEVER HAD A SYSTEMATIC STUDY OF INJURY DEPOLARIZATIONS IN HUMAN BRAIN, SPATIALLY COVERING THE ENTIRE HEMISPHERE, IT'S ELECTRODE STRIPS OR BOLD SIGNAL AND SO FORTH. SO WE DON'T HAVE THAT KIND OF ANALYSIS. MAYBE IN SOME PATIENT WHOSE ARE GENETICALLY SUSCEPTIBLE IT IS CAPABLE OF GOING MUCH FARTHER BUT -- SO I DON'T KNOW THE TRUE STANDARD TO THAT. A MORE SURPRISING FINDING IS THE FACT THAT INJURELY RI DEPOLARIZATION CAUSED SPASM OR VASOCONSTRICTION AND REDUCED TISSUE PROFUSION. AND BACK IN 2006 VASOCON STRICTTIVE NEUROVASCULAR COUPLING, BECAUSE NEUROVASCULAR COUPLING, SOMETHING HAPPENING IN THE BRAIN IN TERMS OF DEPOLARIZATION IS TRIGGERING A VASCULAR EVENT SO IT'S A COUPLING AND IT'S VASOCON STRICTSIVE AS OPPOSED TO NORMAL PHYSIOLOGICAL COUPLING WE ALL TALK ABOUT, FUNCTIONAL OR METABOLIC COUPLING IN BRAIN TISSUE WHERE ACTIVATION OF CERTAIN PART OF THE BRAIN TRIGGERS VASODILATION AND HYPOEMIA IN THAT RESTRICTION OF THE BRAIN TO MAKE UP FOR THE INCREASE ENERGY DEMAND. INJURY DEPOLARIZATION CAUSE VASOCONSTRICTION AND REDUCE PROFUSION TO THAT BRAIN TISSUE. THIS IS THE SAME RAISER SPECKLE MOVIE, THE SAME MODEL AND COLOR MAP. SO NOW WATCH THESE INJURY DEPOLARIZATIONS COMING. THEY'RE ASSOCIATED WITH BLUE WAVE FRONT. THIS IS A SLOWER RUNNING MOVIE. YOU HAVE TO WAIT A LITTLE BIT MORE. THERE'S ANOTHER ONE LOOK AT THE BLUE EXTENT SO THEY CAUSE VASOCONSTRICTION. COLOR CONTRAST WAS NOT GOOD NUMBER TO APPRECIATE THAT BUT MAYBE THIS WILL HELP, THIS IS A MOVIE FROM THE RATS. BLUE IS DECREASE, RED IS INCREASE IN BLUE FLOW AND THIS IS THE WAVE, THIS IS THE DEPOLARIZATION. LET'S WATCH IT AGAIN. THIS IS WHERE DEPOLARIZATION HAPPENS. HERE DEPOLARIZATION IS ALREADY RECOVERED. SO IT CAUSES VASOCONSTRICTION.6 (OFF MIC) >> NOT ALWAYS. THIS IS TWO SLIDES AFTER THIS. SO THIS IS AN EXAMPLE FROM HUMAN. NOW, THIS IS A VERY BUSY SLIDE AND I HATE (INAUDIBLE) FOR THIS BUT THE REAL IMPORTANT TRACING HERE ARE THE RED ONES. THESE ARE RAISER DOPPLER OCTODES INCORPORATED INTO THE ELECTRODE STRIP. THE SAME STRIP I SHOWED HAS A SIX LEAD ACCORDING RAISER DOPPLER FLOW. AS INJURY DEPOLARIZATION HAPPEN THERE'S A DIP IN FLOA. BECAUSE EVERYTHING IS COMPRESSED SO MUCH YOU DON'T APPRECIATE, BUT LOOK AT THE SCALE THE 100% SCALE IS THIS BIG. SO THIS IS ACTUALLY LIKE A 40% REDUCTION IN TISSUE PROFUSION AS THE INJURY DEPOLARIZATION HAPPENS. THE SECOND IN THE SAME PATIENT DETECTED A MUCH BIGGER MUCH WIDER HYPOPER FUSION WHICH IS AGAIN, A BIG ENOUGH TO BE CLINICALLY SIGNIFICANT. HERE, ANOTHER PATIENT SHOWS THE SAME HIGH PRO FUSION THERE. IN HUMAN BRAIN AS WELL THESE CAUSE VASOCONSTRICTION, IT'S NOT JUST ABOUT RODENTS. NOW, WHEN WE WERE PLAYING WITH THE LASER SPECKLE WHICH IS A HIGH SPATIAL RESOLUTION AND TEMPORAL RESOLUTION IMAGING OPTICAL IMAGE, NICE, HANDY WE WERE PLAYING WITH THIS IMAGING THE NEUROVASCULAR EVENTS DURING FOCAL ISCHEMIA. THIS IS A TYPICAL RECORDING IN THIS SETTING. VERTICAL ACCESS BLOOD FLOW CHANGES TAKE PLACE IN THREE DIFFERENT REGIONS OF INTEREST. ONE IS CORE, THE OTHER SOMEONE NON-ISCHEMIC CORTEX. TIME ZERO RIGHT THERE IS MCA OCCLUSION. THERE TH IS WHERE DEFOR LAIZATION HAPPENS. REPETITIVELY AND EACH CAUSES A BLOOD FLOW (INAUDIBLE) BUT WHAT WAS INTERESTING IS THAT THE SHAPE OF THE TRANSIENT IS DIFFERENT DEPENDING ON WHERE YOU PUT YOUR REGION OF INTEREST, THE CLOSER TO THE CORE THE LESS HYPOEMIA YOU GET. SO MILD BRAIN ISCHEMIC TISSUE THE BLOOD FLOW RESPONSE IS HYPOEMIA. THEY DON'T CAUSE -- THEY CAUSE VASODILATION IN THIS ISCHEMIC REGION OF INTEREST. OKAY? EVEN HERE, IF YOU AVERAGE THE AREA UNDER THE CURVE HERE AND THERE YOU MAY BREAK EVEN. SO OVERALL THERE'S NO NET LOSS. BUT IN PENUMBRA THERE'S A MONOPHASIC HYPOPRO FUSION IN EACH WAVE AND IN CORE YOU SEE A DISTANT REFLECTION OF THAT. WHAT'S MORE INTERESTING THOUGH IS IF YOU PAY -- (OFF MIC) EXACTLY WHAT I'M GOING TO SAY NOW. IF YOU LOOK AT THIS, THERE'S LIKE A STEP WISE REDUCTION IN PRO FUSION IN EACH -- IF WE DID NOT HAVE ANY PERIPOLARRIZATION HERE WHAT WOULD THE TREND LOOK LIKE LOOKING DOWN? IS IT SUPER IMPOSITION OR IS THERE ASSOCIATION WITH IT. LET ME -- WHAT TIME IS IT? I'M GOING TO SKIP THAT RAT EXAMPLE AND START THIS. SO TO REITERATE THE ANSWER TO YOUR QUESTION, IF YOU'RE -- SO THIS IS A GRAPH SHOWING BLOOD FLOW PROFILE FROM NON-ISCHEMIC CORTEX INTO THE CORE. THIS IS HYPOTHETICAL, PURELY HYPOTHETICAL T SENOR IS THE CORE, THIS IS THE PENUMBRACH THIS IS THE TYPICAL BLOOD FLOW GRADIANT YOU GET IF YOU DRAW A LINE ON RAISER SPECKLED FORMAT. IN THE MILD IS ISCHEMIC OR NON-ISCHEMIC BRAIN IT CAUSES A NON-HYPEREMIA. IT BECOMES A BIPHASIC RESPONSE. AS YOU GET DEEPER IT BECOMES A MONOPHASIC HYPOPROFUSION BUT RECOVERS COMPLETESLY. AT THE CORE PENUMBRA JUNCTION THE RECOVERY IS OFTEN INCOMPLETE. SO YOU VASOCONSTRICT AND YOU START RECOVERING EVEN IF DEPOLARIZATION IS GONE YOU HAVE A RESIDUAL LOSS IN PROFUSIONCH HEARSAY THE INTERPRETATION OF THE BIG ONE EXAMPLE YOU ASKED ABOUT. I'M GOING TO PROVE THAT TO YOU. TO SHOW WHETHER THIS IS A TRUE RELATIONSHIP OR NOT WE DECIDED TO MEASURE THE AREA OF PROFUSION DEFECT. THE PREVIOUS EXAMPLES WERE REGIONS OF INTEREST PLACED IN CERTAIN REGIONS IN THE BRAIN, A SINGLE POINT MEASUREMENT OVER TIME. WHAT WE DID IN HERE IS USE A THRESHOLD PARADIGM TO IDENTIFY PIXELS WITH A LESS THAN A CERTAIN AMOUNT OF RESIDUAL BLOOD FLOW, IN THIS CASE 20%. SO ALL THAT YOU SEE IN THIS MOUSE HEMISPHERE WITH ON TIEWTION IS ALL THE BLUE PICTURES ARE THOSE WITH LESS THAN 20% RESIDUAL FLOW. WE KNOW THE DIMENSIONS OF THE FIELD, IMAGING FIELD SO WE CAN CALCULATE THE AREA OF THE BLUE REGION. THEN BECAUSE WE'RE IMAGING LIKE AT 1 HERTZ THROUGHOUT AN RO-2 WE CAN PLOT THAT AREA OVER TIME. IT CAN BE A GRAPH OF AREA OF PROFUSION DEFECT. THIS IS A -- HOW IT LOOKS LIKE BEFORE A REPRESENTATIVE PERIIMPRACTICAL ORGANIZATION. HERE IS HOW IT LOOKS LIKE THAT THRESHOLD AREA AFTER DEPOLARIZATION EXPANDED. NOW, IF WE PUT OUR REGION OF INTEREST HERE WE'RE DECKING A MONOPHASIC HYPERPROFUSION THAT WILL RECOVER TO BASELINE. BUT IF YOU PUT A REGION OF INTEREST RIGHT THERE, WHICH BECAME PERMANENTLY BLUE AFTER THE WAY PAST, IT WOULD BE UNDER THE 20% THRESHOLD. IF YOU LOOK AT THE GRAPH, THIS IS WHAT HAPPENS IN THIS ANIMAL. FIRST IT MAKES A BIG JUMP AROUND THE CORE DEPOLAR RISES WITHIN TWO OR THREE MINUTES OF OCCLUSION OF MVCA, THAT'S A PERMANENT SPRING DEPRESSION, PERMANENT INJURY DEPRESSION THAT CAUSES THE SAME VASOCONSTRICTION SO THAT EXPEND PROFUSION DEFECT MORE THAN TWOFOLD, ALREADY RIGHT THERE. WE CAN'T SEE THAT IN HUMANS. BY THE TIME THE PATIENT COMES IT'S OVER THERE, RIGHT? THE TIME SCALE. THIS IS THE FIRST TWO MINUTESCH THIS IS OFTEN MISSED IN CLINICAL STUDIES AND MANY OTHER STUDIES AS WELL. WHAT EACH DEPOLARIZATION DOES IS A STEP WISE INCREASE IN THE AREA OF THE PROFUSION DEFECT. SO YOU CAN SEE THAT THERE'S AN INCREASE. NOW, YOU CAN SAY THAT IF I IGNORE THOSE SPIKES I COULD STILL IMAGINE UPWARD DRIFT IN THIS AREA. TO TEST THAT STATISTICALLY WHAT WE DID IS MEASURE THE AREA JUST BEFORE PID OR INJURY DEPOLARIZATION TO JUST AFTER THEN MEASURED THIS VERSUS THAT, MEASURED THIS VERSUS THAT THEN WE AVERAGE THE CHANGE ACROSS THE PID IN TERMS OF PERCENT INCREASE AND COMPARED TO CHANGE WHEN THERE'S NO PID BETWEEN THE TWO MEASUREMENTS. WHEN YOU DO THIS, THERE'S A DRAMATIC DIFFERENCE. IGNORE THE FIRST LINE. EACH PID THIS CALCULATION SHOWS EACH EXPAND THE PROFUSION BY APPROXIMATELY 10%. BUT THERE WAS NO PID IN BETWEEN. IT DECREASED BY 5%. THERE WASN'T SPONTANEOUS UPWARD DRIFT. IT WAS A STEP WISE INCREASE, IF YOU SEPARATE THE PID IT TENDED TO GET BETTER THE PRO FUSION FIRST LINE SHOW+ WITH OXYGEN DEPOLARIZATION, THE JUMPS UP THERE THE PROFUSION AFFECT AND THAT'S NOT 100% BASELINE, IT'S 160% INCREASE. TWO AND A HALF FOLD INCREASE. SO OBVIOUS QUESTION IS, CAN WE SOMEHOW INTERVENE? HERE WHAT I'M SHOWING YOU IS THERE IS AN ASSOCIATION -- TEMPORAL ASSOCIATION BETWEEN DEPOLARIZATION AND THE LOSS OF PROFUSION IN THE BRAIN TISSUE. RIGHT? THAT DOESN'T MEAN CAUSATION. MAYBE THE PROFUSION GETS WORSE THERE THAT TRIGGERS THE DEPOLARIZATION. ONE WAY TO SHOW IT IS PHARMACO LOGICALLY INTERFERE WITH DEPOLARIZATION. WE DONE HAVE DIRECT EFFECT THEMSELVES. POTENT INHIBITOR OF SPIN DEPRESSION ITSELF SO THESE ARE BASELINE IMAGES, ONE MINUTE AFTER COMMITTED BEFORE DEPOLARIZATION TOOK PLACE. MK 801, THIS IS AN NDA GLUTAMATE RECEPTOR GLOCKER -- BLOCKER. SO THESE TWO IMAGES ARE ONE CONTROLLED, ONE NK, ONE PRE-TREATED ANIMAL, FIVE MILLIGRAMS PER KILOGRAM BEFORE ISCHEMIA, PROF OF CONCEPT. THIS IS BASELINE IMAGE T BLUE REGION THAT YOU CAN SEE THERE, HOPEFULLY, IS 20% THRESHOLD. EVERY BLUE PIXEL HAS LESS THAN 20% RESIDUAL BLOOD FLOW. THIS IS WHAT HAPPENS IN CONTROL ANIMAL AT 60 MINUTESCH THIS PROFUSION DECKED ONE MINUTE, GREW TO BECOME THIS. AT 60 MINUTES. WHEN YOU TREAT THE ANIMAL WITH MKO 1 IT HELP WITH THAT. THE GROWTH WAS STILL THERE BUT NOT AS MUCH AS THIS. AND THE AVERAGE DATA ACROSS MANY ANIMALS, SHOWS THAT MK 301 PRE-TREATMENT REDUCES THE EXTENT OF OXYDEPOLARIZATION EXPANSION AND THE SECOND EXPANSION. THERE'S NO GROWTH BEYOND DEPOLARIZATION. STABLE DOWN TREND WITH NK-1 TREATMENT BUT CONTROL ANIMAL, THERE'S AN EXPANSION OVER TIME. WE EXTEND THIRD DEGREE TO MULTIPLE OTHER AGENTS THAT ARE KNOWN TO INHIBIT SPIN DEPRESSION, MAYBE THIS IS MK 01. WE DON'T HAVE TO TEST OTHER PHARMACOLOGICAL TOOLS, RIGHT? NMDA RECEPTOR NON-COMPETITIVE AND THAT HAD THE SAME EFFECT BUT IT SHRUNK THE AREA OF PROFUSION DEFECT. THE FINAL AREA. THIS IS CORE ISCHEMIC REGION, ISCHEMIA HERE, THE BAR. (INDISCERNIBLE) ARE BOTH SIGMA 1 AGONISTS AND THEY CURIOUSLY ARE ONE OF THE POTENT INHIBITORS OF SPIN DEPRESSION AS WELL BUT NOTHING TO DO WITH NMDA RECEPTOR AND THEY HAVE THE SAME EFFECT. NON-NMDA GLUTAMATE RECEPTOR ANTAGONIST WHICH DOES NOT BLOCK SPIN DEPRESSION AT ALL HAD NO EFFECT ON THE PROFUSION DEFECT. SO THIS AGAIN, STRONGLY SUGGESTIONS THAT IF YOU CAN INHIBIT INJURY DEPOLARIZATIONS, YOU ACTUALLY NOT ONLY MAKE THE TISSUE ELECTROPHYSIOLOGICALLY BETTER, BUT YOU HELP THE PROFUSION OUT SO THERE IS THE POTENTIAL BENEFIT FROM A VASOCON STRICTTIVE NEUROACTIVE COUPLING STANDPOINT. YES. (OFF MIC) >> ALL THESE ARE INTERPERITONEAL ONE HOUR BEFORE ISCHEMIA. (OFF MIC) >> BEFORE ISCHEMIA, YES. (OFF MIC) >> USELESS. IT'S USELESS. HOWEVER INJURY DEPOLARIZATIONS KEEP HAPPENING IN HUMANS FOR UP TO 7 DAYS. DOESN'T HAVE TO BE STROKE. SUB ARACNAL HEMORRHAGE, TRAUMA, IT'S ALL PUBLISHED. SO YOU MAY BE MISSING THE FIRST FEW HOURS BUT IF YOU HAVE DEPRESSION IN INTENSIVE CARE ENVIRONMENT MANY OF THOSE PATIENTS ARE IN THE ICUs YOU CAN SAFELY INTERVENE THE DRUG AND THE NEXT SLIDE IS AN EXAMPLE OF THAT. THIS IS A ANECDOTAL PAPER, STROKE, USING CET MEAN IN PATIENTS WITH LARGE HEMISPHERIC STROKE. TWO PATIENTS ONLY ANECDOTAL BUT WHAT THEY SHOWED IS THAT AGAIN, NOT SURE YOU CAN SEE IT, LET'S LOOK AT PATIENT 1. THESE LITTLE SPIKES THERE MARK INJURY DEPOLARIZATIONS. IN THIS PATIENT AFTER TWO DAYS BETWEEN DAY 2 AND 4 A FLURRY"[ OF DEPOLAR SAIGS WERE ROR AND PATIENTS CLINICALLY SIGNIFICANTLY DETERIORATED, MUCH LESS AROUSABLE AND DEFICITS WORSEN. THEY DECIDED TO GIVE KETAMINE BECAUSE THEY SAW SO MANY INJURY DEPOLARIZATIONS, LIKE A KNIFE STOPS OCCURRENCE OF DEPOLARIZATIONS AND IF YOU READ THE PAPER, PATIENTS IMPROVED CLINICALLY, STARTED WAKING UP AND DEFICITS REVERSED SO THIS IS A VERY GRAPHIC EXAMPLE. HERE, ANOTHER EXAMPLE CHRONOLOGICALLY IN THE FIRST PATIENT WHEN THEY SAW BETWEEN DAY 3 AND 4 SPIN DEPRESSIONS COMING TO THE SCREEN DETERIORATING, AGAIN THE SAME THING HAPPENED AGAIN. SO HERE IS AN EXAMPLE, KETAMINE IS NOT AN EASY DRUG TO USE OBVIOUSLY BUT IT IS USED. IT CAN BE INSTITUTED IN INTENSIVE CARE UNIT ESPECIALLY IF YOU HAVE A PATIENT WITH A HELP CRANEECTOMY AND SEDATED ANYWAY. SO JUST THROWING OUT THERE POTENTIAL USE. YES. (OFF MIC) >> WE DID NOT LOOK AT THAT. WE DID NOT DO THAT STUDY. MY PREDICTION WOULD BE INCREASE OXYGENATION BECAUSE THE PROFUSION INCREASED BUT I CANNOT ANSWER THAT QUESTION BUT IT'S A GOOD QUESTION. >> FAR AWAY FROM ISCHEMIC LESION, YES. YOU GET HYPEREMIA ACTUALLY. >> RIGHT. (OFF MIC) >> I UNDERSTAND NOW. OKAY. I UNDERSTAND THE QUESTION. PERHAPS BUT THAT MILD ISCHEMIC OR NON-ISCHEMIC BRAIN TISSUE DOESN'T NEED THAT EXTRA OXYGEN TO SURVIVE. THAT TISSUE IS NOT HYPOPER FUSED SO ADDITIONAL HYPEREMIA IN THAT TISSUE, I DON'T KNOW HOW MUCH IT WOULD BE A BENEFIT CONSIDERED A BENEFIT. SO YOU BLOCK THESE INJURY DEPOLARIZATIONS MAYBE YOU'RE WORSENING OXYGENATION ELSEWHERE. IS THAT WHAT YOU'RE ASKING? (OFF MIC) AS INJURY DEPOLARIZATION IS PASSING BY, NOT CONSTANTLY. (OFF MIC) >> IT IS CERTAINLY AN INTERESTING -- I DON'T KNOW HOW TO THINK THROUGH THIS IN TERMS OF OVERALL TISSUE OUTCOME, THOUGH. GOOD THING OR BAD THING OR DOES IT MATTER AT ALL? YES. ANOTHER GOOD QUESTION. NOBODY HAS LOOKED AT SUCH THINGS. TECHNICALLY IT'S DIFFICULT. NOW, WE DID LOOK AT ONE THING YEARS BACK, NEVER PUBLISHED IT, PLACE REGION OF INTEREST ON A PEEL ARTERY IN THE MOUSE, THIS IS OCCLUDED MACA, THERE'S SERE LODGE UK BRANCHS FROM MED LINE TOWARDS THIS, THERE ARE PEER COLLATERAL IN THIS MODEL AND WE PLACED THE REGION OF INTEREST ON IDENTIFIED ACA BRANCHES. IMAGE TO FLOW CHANGES IN THOSE BRANCHES DURING PERIPOLARRIZATIONINGS. THEY DIDN'T REALLY GO DOWN SO THERE WAS NO NEGATIVE EFFECT OF FLOW. THERE WAS NO CONSTRICTION IN THOSE VESSELS BECAUSE THEY WERE OUTSIDE. MILD ISCHEMIC. I CANNOT ANSWER WHETHER THE TRANSIENT HYPEREMIA IN THOSE VESSELS HAS ANY BENEFIT IN TERMS OF IMPROVING THE COLLATERAL OF FLOW INTO ISCHEMIC TISSUE. THERE MAYBE A BENEFIT BUT AGAIN IT'S TWO MINUTES OF INCREASED FLOW, THOSE ARE GOOD QUESTIONS, THIS IS SOMETHING WE DISCUSSED EARLIER. ARE YOU DOING AN OVERALL GOOD OR NOT BY SUPPRESSING THESE INJURY DEPOLARIZATIONS. YOUR IMPROVING PROFUSION IN WHERE ISCHEMIA IS BUT PERHAPS OTHER EFFECTS THAT BENEFIT THE TISSUE. (OFF MIC) >> RIGHT. EXACTLY. >> LOOK AT WHAT HAPPENS OUTSIDE. WE DIP DO THAT. YES, YOU CAN DO THAT. WE DIDN'T DO THAT. I HONESTLY DIDN'T PAY ATTENTION -- IT'S TOO FAR NOW TO SHOW, WE DIDN'T PAY ATTENTION TO THAT UPWARD INCREASE IN FLOW IN THE MILD ISCHEMIC BRAIN THAT MUCH. WE THOUGHT WE DIDN'T -- THAT'S A FAIR QUESTION, IS THAT GOING UP BECAUSE OF INJURY DEPOLARIZATION, SOME LASTING VASODILATOR EFFECT OR NOT. THAT'S A GOOD SPECIMEN. MOVING ON. OKAY. DRUGS ARE NOT THE ONLY IMPORTANT THING, I HAVE FIVE MINUTES TO HIT YOU WITH SOMETHING GREAT FOR MIGRAINE. A PEAK MODULATOR OF SPRAYING DEPRESSION SUSCEPTIBILITY IN THE BRAIN ARE GENES, NOT HERB IS EQUALLY SUSCEPTIBLE TO SPIN DEPRESSION. NOT EVERYBODY HAS WITH THE SAME STIMULUS, THAT'S MOTDLATED BY GENES NEVER EXPERIENCE SUCH A THING IN THEIR LIVES. SO IF SPIN DEPRESSION SUSCEPTIBILITY CAN BE MODULATED GENETICALLY DOES THAT TRANSFER TO INJURE DEPOLARIZATION SUSCEPTIBILITY WHEN THERE'S INJURY IN THE BRAIN. ONE OF THE GENETIC MOUSE MODELS THAT WE HAVE IS A MUTANT MOUSE MODEL FAMILIAR MIGRAINE. WE HAVE SHOWN, THIS MUTATION WHAT IT DOES IS THIS IS A MUTATION C 82.1 CHANNEL, IT SHIFT IT IS VOLTAGE CURVE TO THE LEFT. MEANING CHANNELS ARE OPEN OING AT MORE NEGATIVE MEMBRANE POTENTIALS AND PEAK INCREASE SO OVERALL GAIN OF FUNCTION OR INCREASE EXCITABILITY PHENOTYPE YOU'RE GETTING AT THE CHANNEL LEVEL. AS WELL AS AT THE SPIN DEPRESSION LEVEL. THIS IS AN EXAMPLE WHERE WE TOTALLY APPLY A CONSTANT CONCENTRATION OF POTASSIUM CHLORIDE AND RAPID SPIN DEPRESSIONS. THESE ARE REPETITIVE SPIN DEPRESSIONS. I HAVE A WILD TYPE ANIMAL, MALE AND FEMALE AND R-2 MUTANT IN THE SAME CHANNEL, FOUND IN HUMANS. THE HUMAN MUTATION, NOT AKIN TO THE MOUSE. THIS IS A MORE SEVERE HUMAN MUTATIONCH THESE PATIENTS ACTUALLY BECOME COMATOSE IF THEY -- GET HIT BY A TENNIS BALL IN THE HEAD. THEY JUST DROP AND THEY STAY IN COMA FOR WEEKS AT A TIME. SO IT'S A REMARKABLE DISEASE OR SYNDROME. SO WHAT WITH WE SHOWED A NUMBER IS THESE MUTANT MOUSE MODELS WHERE MOST SUSCEPTIBLE TO SPIN DEPRESSION. IF YOU PUT TOPICAL POTASSIUM CHLORIDE YOU HAVE A MACHINE GUN, AND FEMALES WERE MORE SUSCEPTIBLE MALES WHICH GO TO -- SO USING THESE MICE WE LOOK AT INJURY DEPOLARIZATION OCCURRENCE IN FOCAL ISCHEMIA. REMARKABLY ENOUGH, I I WISH YOU COULD SEE IT A WILE TYPE TRACING IS ON TOP AND SHN-1 HERE, THE SECOND TRACING INJURY DEPOLARIZATION HAPPENS IN THE WILD TYPE ANIMAL DURING IDENTICAL OCCLUSION MODEL. FILAMENT OKAY CLIEWG MODEL AND THE FHM-1 MUTANT WAS FIRING INJURY ARE DEPOLARIZATIONS LIKE A MACHINE GUN. THE POOL DATA IS HERE EACH CIRCLE IS INJURY DEPOLARIZATION, EACH LINE IS ONE ANIMAL. YOU CAN SEE HOW MANY MORE DEPOLARIZATIONS YOU GET IN THE MIGRAINE MUTANT MIESMS HERE IS THE CUMULATIVE DATA, OCCURRENCE OF INJURY DEPOLARIZATION OVER TIME, THIS IS ALL ANIMALS POOLED AND THE CURVES DIVERGE AND YOU HAVE TWOFOLD INCREASE IN FREQUENCY OF INJURY DEPOLARIZATION IN SHN-1 MUTANTS. AND THE AVERAGE NUMBER IS THERE. THIS IS I THINK VERY IMPORTANT, IF YOU'RE A MIGRAINER, MIGRAINE WITH AURA, THAT MEANS YOU ARE MORE SUSCEPTIBLE TO DEVELOP INJURY DEPOLARIZATIONS, IF YOU END UP GETTING THE STROKE. AND YOUR STROKES MAYBE WORSE. (OFF MIC) >> THERE IS A BIG EPIDEMIOLOGIC LITERATURE SHOWING MIGRAINE AND STROKE, THAT'S WHY WE'RE EXCITED ABOUT THE FINDING. ALL THE FOCUS GOES TO VASCULATURE, IF THERE'S SOMETHING WRONG WITH THE VASCULATURE PERHAPS GIVING THE STROKE BUT PERHAPS THE BRAIN IS HYPEREXCITABLE AND -- (OFF MIC) ALSO FREQUENCY OF STROKE. (OFF MIC) >> EXACTLY RIGHT. BUT YOU CANNOT ANSWER THAT QUESTION BECAUSE YOU HAVE STO HAVE STANDARDIZED COMPARISON OF MIGRAINE PATIENT. WE'RE WRITING GRANT AS WE SPEAK ON JUST LOOKING AT THAT, GETTING PATIENTS IN, SEGREGATING THEM, MIGRAINE WITH AURA, MIGRAINE WITHOUT AURA, NO MIGRAINE AND BASELINE MRI AND THAT SHOWS HOW BIG THE IMPACT GOT, THAT HASN'T BEEN DONE. (OFF MIC) >> NO WE DID -- THE OUTCOMES WERE MUCH MORE SEVERE. I DON'T HAVE THE SLICE HERE, BECAUSE OF THE TIME. THE FIVE MINUTES ARE UP. (OFF MIC) LET ME SAY ONE THING, TISSUE OUTCOME IN MIGRAINE MUTANT MICE THE IMPACT WERE DOUBLED. THE MORTALITY WAS DRAMATICALLY HIGHER. WE COULDN'T EVEN USE FOUR DIED, WE ONLY HAD ONE ANIMAL, ONE LITTLE ERROR BAR ON TOP SO IT WAS A MARKEDLY WORSE PHENOTYPE IN THESE ANIMALS. OF COURSE MOST MIGRAINE PATIENTS END UP GETTING A STROKE ARE NOT FHM-1 PATIENTS THAT'S AN EXTREME EXAMPLE BUT IT'S A PROOF OF PRINCIPLE THING. THERE'S INTRINSIC EXCITABILITY IT TRANSLATES TO A LARGER STROKE. AND COMING BACK TO THE THE FREQUENCY OF STROKE SOME OKAY CLIEWGS WILL BE TRANSIENT, WE GET LITTLE OKAY CLIEWGS IN THE BRAIN THAT BECOMES A -- OCCLUSIONS IN THE BRAIN, SOMETIMES WE DON'T PERCEIVE IT BECAUSE THE REDUCTION IS MILD BUT IF YOU'RE HYPEREXCITABLE AND MORE TONAL SUSCEPTIBLE TO DEPOLAR RISE YOU WILL PROCEED AS A SYMPTOM. IF IT'S A LITTLE WORSE, YOU WILL ENUP GETTING A STROKE RATHER THAN A TIA. THE FREQUENCY GOES UP AS WELL FROM THAT POINT OF VIEW. (OFF MIC) >> THAT'S CALLED MIGRAINE INFARCTION. THAT'S A DIFFERENT ENTITY. (OFF MIC) >> I DON'T KNOW HOW THAT HAPPENS. THAT'S A GOOD QUESTION. VASOCONSTRICTION? YES. THAT'S ONE OF THE POSSIBLE MECHANISMS. THE OTHER MECHANISM IS THEY DID HAVE A REAL STROKE. BUT THEY PRESENTED LIKE A MIGRAINE ATTACK. SO WHEN THESE PATIENTS PRESENT IN THE EMERGENCY ROOM TO SAY HOW DID IT START, I DON'T KNOW I WAS EATING AND HAD A FUNNY FEELING AND THIS AND THAT. IF YOU DON'T KNOW IF IT WAS A PARTIAL OCCLUSION THAT CLOGGED UP AND BECAME COMPLETE OCCLUSION SO IT GIVES YOU THE PERCEPTION OF PROGRESSIVE DEFICIT LIKE A MIGRAINE AURA, VERSUS IF IT WAS A TRUE MIGRAINE AURA IT ENDS UP INFARCTING THE TISSUE. CAUSE AN EFFECT IS VERY DIFFICULT IN MIGRAINE INFARCTION. WE EXTEBBED THE SAME DATA -- EXTENNED THE SAME DATA TO (INAUDIBLE) MICE. (INDISCERNIBLE) PEOPLE FOCUS ON THE SMALL VESSEL DISEASE, BUT THE FIRST SYMPTOM OF (INDISCERNIBLE) IS MIGRAINE WITH AURA, THAT STARTS IN LATE TEENS, EARLY 20s. SEVERE MIGRAINES WITH AURA, AS THE STREAKS KICK IN, MIGRAINE DISAPPEARS SO THIS IS A MIGRAINE SYNDROME ESSENTIALLY. WE LOOK AT THE SPIN DEPRESSION SUSCEPTIBILITY IN THESE ANIMALS FIRST AND FOUND INCREASE, SO THIS IS THE WILE TYPE, THIS IS THE MUTANT AND THIS IS THE THRESHOLD OF SPIN DEPRESSION, WE INCREASE DOWN UNTIL WE GET THE SPIN DEPRESSION, THAT WAS THE THRESHOLD AND IT WAS MUCH LOWER. SO THESE ANIMALS ARE MORE SUSCEPTIBLE TO SPIN DEPRESSION. AND THEY DEVELOP MORE INJURY DEPOLARIZATIONS COMPARED TO WILD TYPE MCH MODEL. THIS IS THE POOLED DATA AND THIS IS MUTANT, THIS IS THE WILD TYPE. SO HERE IS A COMPLETELY DIFFERENT HUMAN DISEASE. IF MUTATIONS ARE IN KNOWLEDGE 3, IT'S A MEMBRANE RECEPTOR DETERMINING CELL FATE DETERMINATION BASED ON CONTIGUOUS, THERE'S NOWHERE ELSE. BUT THESE BRAINS ARE HYPEREXCITABLE YET THEY SHOW AND THEY SHOW THE SAME INJURY DEPOLARIZATION PHENOTYPE. NOW, I'M GOING TO FINISH UP IN THREE MINUTES BY SHOWING YOU SOMETHING WITH DATA. THIS IS VERY EARLY PRELIMINARY DATA, DON'T SCORN US FOR HAVING SHOWN BUT IT'S SO EXCITING THAT I COULDN'T HELP. SO HOW CAN WE GET TO THIS QUESTION? THE QUESTION YOU ASKED IS A VERY IMPORTANT ONE. DOES THAT REALLY MAKE THE INFARCT WORSE? ONE WAY TO ANSWER THIS STO LOOK AT DIFFUSION PROFUSION MISMATCH. IF YOU HAVE THIS BIG PROFUSION DEFECT ON MR, ACUTE STROKE MR, AND THE IMINFARCT, THERE'S A BIG MISMATCH. SO ONE WAY TO LOOK AT HOW SUSCEPTIBLE THE BRAIN IS TO DEPOLARIZATION IS DIFFUSION MISMATCH. SO IN A BRAIN THAT IS MORE SUSCEPTIBLE TO DEPOLARIZATION, TO DEPOLARIZE AND CONVERT THEIR PENUMBRA INTO CORE YOU EXPECT LESS MISMATCH WHEN YOU SCAN THEM. THE ENTIRE PROFUSION DEFECT DEPOLARIZES. HAVING MORE MISMATCH SUGGESTS THAT THE TISSUE IS NOT AS SUSCEPTIBLE TO DEPOLARIZE, THAT'S THE IDEA. SO WE LOOKED AT THIS IN A SERIES OF 1800 PATIENTS, A RETROSPECTIVE SCAN OF OUR DATABASE. 740 HAD MRI WITHIN 48 HOURS AND THEN ONLY 68%, 10% OF THEM, 68 PATIENTS HAD MIGRAINE STATUS DOCUMENTED IN THE CHARTS, SO THAT TOOK OUT A BIG CHUNK. WE HAD 68 TO WORK WITH. THERE WAS NO PROFUSION DEFECT IN 28 SO WE DOWN DO THE PROFUSION DIFFUSION MISMATCH ANALYSIS. SO WE WERE LEFT WITH 40 PATIENTS. HERE IS THE BREAK DOWN. IN PATIENTS WITH MYGRAIN -- IN PATIENTS WITH MIGRAINE WITH AURA HISTORY. I HAVE TO ORIENT MYSELF HERE. 50% OF PATIENTS WITH MIGRAINE WITH AURA, THAT'S ONLY SIX PATIENTS, HAD NO DIFFUSION PROFUSION MISMATCH. NO DIFFUSION PROFUSION MISMATCH. THE ENTIRE EFFECT IS DIFFUSION BRIGHT. HALF OF THE PATIENTS HAD THAT. ONLY 14% OF THE PATIENTS WITHOUT AURA BUT WITH MYGRAIM GRAIN HAD THAT FINDING AND NONE OF THE 27 PATIENTS THAT HAD NO MIGRAINE HISTORY HAD NO PROFUSION DIFFUSION PROFUSION MISMATCH. SO MIGRAINE WITH AURA PATIENTS HAVE HIGH INCIDENCE OF COMPLETELY DEPOLARIZING OR MAKING THEIR PROFUSION DEFECT THE CORE. THAT'S SOMETHING THAT WE'RE NOW BUILDING ON, THE EXTENT -- WE EXTENDED OUR STUDY TO 3,000, 6,000 PATIENTS HOPING TO GET MORE NUMBER IN HERE TO AT LEAST GET A HANDLE ON WHAT YOU ASK. DOES THIS EXPAND THE DEFICIT, YES. (OFF MIC) THIS IS RETROSPECTIVE. IT'S NOT ONLY 3%, IT'S 50%, THIS IS ONLY THREE PATIENTS BECAUSE OF ALL THE KICKED OUTPATIENTS. SO WE ONLY HAD 40 PATIENTS TO ANALYZE SO 3 OUT OF 40. 'S STILL A HUGE NUMBER BUT IF YOU DO THE STUDY IN 'LIABLE WAY, MAYBE PATIENTS DOCUMENTED WITHOUT AURA MAY HAVE HAD AN AURA SOMETIME, THE DISTINCTION BETWEEN WITHOUT AURA AND WITH AURA IS SOMETIMES (INAUDIBLE). SO I THINK I WILL LEAVE -- WELL, I CAN'T WITHOUT SAYING THIS. MIGRAINE PROPHYLAXIS. WE HAVE SHOWN BACK IN 2006 THAT MIGRAINE PROPHYLAXIS ACTUALLY DEPRESSES YOUR -- SUPPRESSES YOUR SUSCEPTIBILITY TO SPIN DEPRESSION, MAKES IT LESS LIKELY YOUR BRAIN, CROBBIC TREATMENT WITH -- (INDISCERNIBLE). AND WE TESTED THEM OOD WAY OF -- ANOTHER WAY OF SUPPRESSING SPIN DEPRESSION ON PID FREQUENCY, THEY BOTH REDUCE (INAUDIBLE) AND THE INFARCT -- SO THERE IS (INAUDIBLE) TO ENTERFEE. THE NEXT OBVIOUS THING TO DO IS DURING THE PROSPECTIVE STUDY TO DOCUMENT THOSE PATIENTS WHO ARE ON MIGRAINE PROPHYLAXIS AND LOOK AT WHETHER THEY DEVIATE FROM THOSE MIGRAINE WITH AURA PATIENTS WHO ARE NOT ON MEDICATIONS AND JUST DEMONSTRATE THAT. YES. (OFF MIC) YEAH, NO, NO, WE TESTED AMITRIPTYLINE, THAT SUPPRESSES NOT INJURY BUT SPIN DEPRESSION SUSCEPTIBILITY. WE TESTED A BETA BLOCKER, THAT SUPPRESSES SPIN DEPRESSION SUSCEPTIBILITY AND (INDISCERNIBLE) WHICH IS ABANDONED OLD PERFECTED DRUG, THAT ALSO DOES THE SAME THINGCH THOSE THREE DRUGS WE DIDN'T TEST THE PID WE PICKED THE TWO DRUGS. AND BY THE WAY (INDISCERNIBLE) IS ANOTHER ONE THAT DOES THE SAME THING. IT'S KIND OF A CROSS ACTION FOR MIGRAINE DRUGS IN A SENSE WE'RE GOING TOWARD THAT NOW, WE'RE GOING TO PICK A DRUG UNRELATED IN (INAUDIBLE) MODEL AND SEE IF THEY ALSO ARE CAPABLE OF SUPPRESSING PID. >> YES. (INAUDIBLE) I BELIEVE ONE MAJOR MECHANISMS OF ACTION IS GLUTAMATE RELEASE REDUCTION. (OFF MIC) >> GOOD. (OFF MIC) >> RIGHT. S THAT POST SYNAPTIC ACTION YOU'RE TALKING AB, NOT THE (INAUDIBLE). RIGHT. OKAY. SO GOOD TO KNOW THAT. (INDISCERNIBLE) IN OUR HANDS IS REALLY POTENT IN SUPPRESSING THE INJURY DEPOLARIZATION, THAT'S LESS THAN ONE PER HOUR, THAT'S A REMARKABLE REDUCTION. I WANT TO STOP NOW. SO THIS IS MY LARGE GROUP NOW. THESE ARE CURRENT MEMBERS, THESE ARE DISEASE MEMBERS AND I HAVE ALL THE CONTRIBUTORS, I CAN'T GO THROUGH THEM ONE BY ONE BUT I THANK THEM, THANK YOU FOR LISTENING. [APPLAUSE] (OFF MIC) YES. EXACTLY RIGHT. (OFF MIC) WE HAVE AN ENERGY DEFICIT THAT SPREADS OUT OF THE ISCHEMIC REGION AND THE DEGREE OF RECONSTITUTION OF ATP DETERMINE THE EXTENT OF THE (INAUDIBLE). >> THAT'S EXACTLY RIGHT. SO WHERE IT MATTERS WHERE DOES PARTIAL ISCHEMIA, VERSUS THE MISMATCH? WRIT MAY NOT MATTER, MILD ISCHEMIA OUTSIDE THERE, MAYBE A POSITIVE EFFECT. BUT I DON'T KNOW IF THAT'S CLINICALLY RELEVANT. SO YES. MAKES THE MISTACH WORSE BY INCREASING METABOLIC DEMAND AND REDUCING SUPPLY BECAUSE OF VASOCONSTRICTION AROUND ISCHEMIC REGION. >> IS THERE A ROLE FOR METABOLITES FOR ARACHIDONIC ACID LIKE PROSTAGLANDIN (INAUDIBLE) THAT REGULATES BLOOD FLOW (INAUDIBLE) >> I DON'T KNOW. >> DEPOLARIZATION (INAUDIBLE) OF THE ENZYMES. >> I DON'T KNOW THE ANSWER. EVERYBODY HAS LOOKED AT IT. I KNOW THE LITERATURE PRETTY WELL BUT I DON'T KNOW. >> (INAUDIBLE) RELEASED DURING EPILEPSY >> WHAT DO I THINK IS DOING THE -- (OFF MIC) (OFF MIC) >> RIGHT. RIGHT. RIGHT. SO THE THE LEADING CANDIDATE IN MY MIND IS POTASSIUM ITSELF. (OFF MIC) >> YEAH. POTASSIUM -- 40 MILLIMOLARS IF YOU DUMP 40 MILLIMOLAR PIPETTE, IT'S MEMBRANE CHANNELS OPEN THE CAPSID DEPOLARIZE AND DECONSTRICT. (OFF MIC) >> THERE IS A DIFFERENCE. RIGHT. (OFF MIC) >> SO HERE, EXACTLY RIGHT. THAT'S TUG OF WAR BETWEEN VASODILATOR INFLUENCE AND VASOCONSTRICTOR INFLUENCE DURING DEPOLARIZATION RATE. SO WHAT WE HAVE DONE IN THE PAST IS REDUCE BLOOD PRESSURE ARTIFICIALLY BUT WITHDRAWING BLOOD SO THE INTRAVASCULAR PROFUSION PRESSURE WAS REDUCED. BUT VASODILATION KEPT THE BLOOD FLOW RELATIVELY CONSTANT. SO WE DROPPED BLOOD PRESSURE TO 45-MILLIMETERS, AND BLOOD FLOW WAS REDUCED BY 10%, 15% OR SO, SO BLOOD FLOW IS MAINTAINED. WE INDUCE SPIN DEPRESSION ON TOP OF THAT. THAT CAUSES VASOCONSTRICTION EVEN IN NORMAL BRAIN. SO IF YOU DON'T HAVE PRESSURE SUPPORTING IT, EVEN THAT IS IMPORTANT TO DETERMINE WHETHER YOU CONSTRICT THE TUG OF WAR IS SO CLOSE BY PUSHING ONE WAY OR THE OTHER YOU CAN IB VERT THE RESPONSE FROM VASODILATION TO VASOCONSTRICTION. (OFF MIC) >> GREAT QUESTION. YES, SO INJURE I DEPOLARIZATION PLUT TISSUE AND GLUTAMATE AND EVERYTHING ELSE, AND IT'S ALMOST CERTAIN THAT THE PERIVASCULAR NERVE, PERIVASCULAR NERVES ARE BS BAITENING A LOT OF POTASSIUM AND THEY RELEASE EVERYTHING THEY HAVE GOT. THAT'S ONE OF THE NEUROGENIC INFLAMMATION HYPOTHESIS OF MIGRAINE HEADACHE, THE VASOACTIVE PEPTIDES ARE TRIGGERED AND THAT GIVES YOU THE HEADACHE. THAT'S THE IDEA. (OFF MIC) >> ONE GROUP FROM GERMANY IF YOU WANT TO LOOK UP THIS PAPER, JCBF DID THAT ABOUT TEN YEARS AGO, 12 YEARS AGO OR SO, UNFORTUNATELY I DON'T REMEMBER WHAT HE FOUND. HE INNERVATED SPINNING DEPRESSION IN OTHERWISE NORMAL BRAIN, NOT STROKE, BUT OTHERWISE NORMAL BRAIN. HE FOUND A DIFFERENCE, I DON'T REMEMBER IF SRI SEW DILATION WAS DIMINISHED OR VE VERSED. I HAVE TO LOOK T IT UP TO REMIND MYSELF. (OFF MIC) >> THE QUICK ANSWER IS NO. BUT WHAT WE HAD DONE, ANOTHER SERIES OF EXPERIMENTS WE ARE DOING NOW IS ZINC IMAGING, YOU'RE USING FLOWS IN 3 WHICH IS A ZENC FLUROPHORE. ZINC IS CONCENTRATED AT HIGH LEVELS IN PRE-SYNAPTIC TERMINALS. AND TRANSPORTED WITH GLUTAMATE. WHEN THERE'S GLUTAMATE RELEASE AT THE SAME TIME THERE'S MASSIVE ZINC RELEASE IN THE EXTRA CELLULAR SPACE, IT'S TAKEN UP QUICKLY. SO USE THIS AS A SURROGATE MARKER OF GLUTAMATE RELEASE WE STARTED DOING ZINC IMAGING TO LOOK AT GLUTAMATE IMAGING BUT ZINC FLUROPHORE IS ALSO FOR CALCIUM SO WE HAVE TO USE EDPA SO THE IN VIVO CRYOWINDOW, THE SURFACES (INDISCERNIBLE) AND THAT'S THE (INAUDIBLE) THAT MIGHT ANSWER YOUR QUESTION. THE SPIN QUESTION IS NOT MODIFIED. SO IT -- I DON'T KNOW WHAT HAPPENS TO BLOOD FLOW RESPONSE BUT SPIN DEPRESSION IS NOT MODIFIED. (OFF MIC) >> IT'S -- IT'S INTRAPERITONEAL INJECTION. (OFF MIC) >> WITHIN TEN MINUTES YOUR DOSE IS IN THE BRAND T ANIMAL CHANGES ITS BEHAVIOR. BEHAVIORAL SIDE EFFECTS SO YOU CAN SEE THE BEHAVIOR IS QUIET AND SO IT'S FREELY ENTERING THE BRAIN. ONE LAST QUESTION MAYBE, YEAH. (OFF MIC) >> YES. THE ANSWER IS YES. WHEN YOU DO ISCHEMIC PRE-CONDITIONING YOU'RE TRIGGING ALSO THE INJURY DEPOLARATIONS NOT TO THE EXTENT THEY DAMAGE THE BRAIN. SO THEY DO HAVE BENEFICIAL EFFECTS ON THE TISSUE ITSELF. THEY TRIGGER REACTIVE ASTRO CYTOSIS. REACTIVE ASTRO SITES ARE VERY PROTECTIVE IN STROKE. SO IF YOU HAVE THAT IN BRAIN YOU GET A SMALLER IMPACT, WE SHOW THAT, TESTED THAT AND SHOWED THAT. THAT'S ONE WAY. THE SECOND THING IS I FORGOT THE SECOND THING I WAS GOING TO SAY. SORRY. (OFF MIC) >> OKAY. THANK YOU.