1. Project ABLADE “Advanced Bladder cancer LAser Diagnostics and thErapy” University of Dundee, Nethergate, Dundee, DD1 4HN, Scotland, UK SPE “LAZMA”, Techpark Strogino, Moscow, 123458, Russia EVALUATING ADAPTATION OPTIONS MICROCIRCULATORY-TISSUE SYSTEMS BASED ON CORRELATION NUTRITIVE BLOOD FLOW AND REDOX RATIO. Simian Zhu, University of Dundee, UK

2. Project ABLADE “Advanced Bladder cancer LAser Diagnostics and thErapy” University of Dundee, Nethergate, Dundee, DD1 4HN, Scotland, UK SPE “LAZMA”, Techpark Strogino, Moscow, 123458, Russia Laser Doppler Flowmetry The dynamic changes of blood perfusion can be recorded by Laser Doppler Flowmetry (LDF) and analysed by mathematic tools; considering the physiology of microvascular, the frequency spectrum of LDF signal after wavelet transfer can be corresponded to five rhythms(Dunaev, Sidorov et al. 2013): RhythmsFrequency interval (Hz) Endothelial0.0095-0.02 Neurogenic0.02-0.06 Myogenic0.06-0.16 Breathing0.16-0.4 Pulse (Cardiovascular)0.4-1.6

3. Project ABLADE “Advanced Bladder cancer LAser Diagnostics and thErapy” University of Dundee, Nethergate, Dundee, DD1 4HN, Scotland, UK SPE “LAZMA”, Techpark Strogino, Moscow, 123458, Russia Fluorescence & Redox-ratio The fluorescence data were measured by pairs; the backscattered light from tissue was separated through spectrometer, received by computer, and displayed in wavelength-intensity graph. In most instances, there are two peaks in the graph, one at the short wavelength is the excitation laser peak (I ex ), and another at the longer wavelength is the fluorescence peak generated by tissue fluorophores (I em ). The intensity of each peak was then recorded for calculation of redox ratio. In biological tissues, there are various components that can absorb the light, which can largely affect the fluorescence spectra. In order to reduce the affection of tissue absorption, we tried four different ways to calculate the redox ratio:

4. Project ABLADE “Advanced Bladder cancer LAser Diagnostics and thErapy” University of Dundee, Nethergate, Dundee, DD1 4HN, Scotland, UK SPE “LAZMA”, Techpark Strogino, Moscow, 123458, Russia Redox-ratio

5. Project ABLADE “Advanced Bladder cancer LAser Diagnostics and thErapy” University of Dundee, Nethergate, Dundee, DD1 4HN, Scotland, UK SPE “LAZMA”, Techpark Strogino, Moscow, 123458, Russia Occlusion and Reactive Hyperemia By the applying pressure on upper limb (>50 mm Hg than systolic blood pressure), the blood flow within the Brachial artery can be blocked, such process called occlusion or artificial ischemia. The perfusion and venous return are blocked on the occlusion site, with the consumption of oxygen, the metabolism level will change and can be reflected in the Redox-ratio. With pressure disappear, the perfusion fast recovered and largely increase to exceed the normal level (Reactive Hyperemia). After while the perfusion gradually back to the normal level (Recovery), during this period, the metabolism and nutritive blood (Imn) may have some correlations.

6. Project ABLADE “Advanced Bladder cancer LAser Diagnostics and thErapy” University of Dundee, Nethergate, Dundee, DD1 4HN, Scotland, UK SPE “LAZMA”, Techpark Strogino, Moscow, 123458, Russia Experiment design Blue and UV fluorescence LDF LAKK-M on fluorescence channel, LAKK-0P on LDF channel, fibre probe from each device adhere on the skin parallel using medical tape. Fluorescence excitation wavelength: UV: 365nm Blue: 450nm There are 4 stages in the experiment: State of rest (S1): 3min Occlusion (S2): 3min, 220mm Hg pressure on upper limb Reperfusion (S3): 3min Recovery (S4): 21min, fluorescence measure every 3min in Stage 4

7. Project ABLADE “Advanced Bladder cancer LAser Diagnostics and thErapy” University of Dundee, Nethergate, Dundee, DD1 4HN, Scotland, UK SPE “LAZMA”, Techpark Strogino, Moscow, 123458, Russia Wavelet Transform Analysis The frequency spectrum of LDF signal was obtained by applying wavelet transform. As described previously, there are five frequency interval correspond to rhythms, and the myogenic rhythm is the main target we are focus on. Using fragment tool in LAKK-M analysis software to divide the signal into fragments based on the stages and apply wavelet transform on each fragment respectively.

8. Project ABLADE “Advanced Bladder cancer LAser Diagnostics and thErapy” University of Dundee, Nethergate, Dundee, DD1 4HN, Scotland, UK SPE “LAZMA”, Techpark Strogino, Moscow, 123458, Russia Results Three volunteers age 50±16. All the volunteers show the similar Redox-ratio trends during S1-S4 : Decreasing metabolism during occlusion, increasing during reactive hyperemia, and back to normal level in recovery stage. Different volunteers have different speed of recovery, from the data provided here, there are no significant correlation between speed and ages. (Age 66: 12min, age 50: 9min, age 34: 15min, p<0.05)

9. Project ABLADE “Advanced Bladder cancer LAser Diagnostics and thErapy” University of Dundee, Nethergate, Dundee, DD1 4HN, Scotland, UK SPE “LAZMA”, Techpark Strogino, Moscow, 123458, Russia Results Nutritive blood (Imn) is calculated from total blood perfusion, it presents the nutritive blood volume in target tissue, which has closer relationship with the metabolism. In stage 2, regarded as no blood flow within target tissue. Volunteers have less Imn in S3 than in S1, while the Imn increased largely after 3min of occlusion and continually adjusting during the Recovery stage. The younger volunteer has significant higher nutritive blood perfusion than other two elder volunteers (p<0.01)

10. Project ABLADE “Advanced Bladder cancer LAser Diagnostics and thErapy” University of Dundee, Nethergate, Dundee, DD1 4HN, Scotland, UK SPE “LAZMA”, Techpark Strogino, Moscow, 123458, Russia Analysis

11. Project ABLADE “Advanced Bladder cancer LAser Diagnostics and thErapy” University of Dundee, Nethergate, Dundee, DD1 4HN, Scotland, UK SPE “LAZMA”, Techpark Strogino, Moscow, 123458, Russia Conclusion By normalize the Redox-ratio, the link between the change rate of metabolism during the occlusion and reactive-hyperemia can be established. Preliminary data shows there are linear correlations (r=0.92564) between the change rate during Stage 2 and Stage 4, which means the volunteers with more Redox-ratio decrease during occlusion having better adaptation and faster recovery from reactive-hyperemia.

12. Project ABLADE “Advanced Bladder cancer LAser Diagnostics and thErapy” University of Dundee, Nethergate, Dundee, DD1 4HN, Scotland, UK SPE “LAZMA”, Techpark Strogino, Moscow, 123458, Russia Discussion The preliminary conclusion of link between the adaptation of options of MTS shows a remarkable linear correlation, however, it still remains more experimental data support. At present, the fluorescence device has to be manual operated, which may leads to unnecessary vibration and slow sampling speed, therefore a device optimizing is needed.

13. Project ABLADE “Advanced Bladder cancer LAser Diagnostics and thErapy” University of Dundee, Nethergate, Dundee, DD1 4HN, Scotland, UK SPE “LAZMA”, Techpark Strogino, Moscow, 123458, Russia Acknowledgment Project ABLADE “Advanced Bladder Laser Diagnostics and thErapy” was funded by FP7 funding Grant Agreement No: 324370

14. Project ABLADE “Advanced Bladder cancer LAser Diagnostics and thErapy” University of Dundee, Nethergate, Dundee, DD1 4HN, Scotland, UK SPE “LAZMA”, Techpark Strogino, Moscow, 123458, Russia Thank you!

15. Project ABLADE “Advanced Bladder cancer LAser Diagnostics and thErapy” University of Dundee, Nethergate, Dundee, DD1 4HN, Scotland, UK SPE “LAZMA”, Techpark Strogino, Moscow, 123458, Russia EVALUATING ADAPTATION OPTIONS MICROCIRCULATORY-TISSUE SYSTEMS BASED ON CORRELATION NUTRITIVE BLOOD FLOW AND REDOX RATIO. Simian Zhu, University of Dundee, UK