Laser Doppler Flux Research Articles

Relationship Between Finger Photoplethysmographic Pulses and Skin Blood Perfusion.

Introduction Photoplethysmography (PPG) measures are important in monitoring peripheral oxygen saturation (SpO2). Another parameter is a derived quantity referred to as the peripheral perfusion index (PPI). It is calculated as the ratio of the peak-to-peak pulse amplitude of a PPG signal (PAPPG) to the non-pulsating part of the total PPG signal. The PPI has been used as a marker of blood perfusion states in a variety of clinical settings but has not been systematically and directly compared to measures of local blood perfusion. This study's purpose was to investigate this issue to provide initial data on the relationship between finger skin blood perfusion, measured by laser Doppler blood perfusion flux (LDF) and PAPPG. Methods Ten subjects (five male), recruited from medical students with an average age of 26 years, participated.While supine for 30 minutes, skin blood perfusion was recorded using laser Doppler flux (LDF) on the ring finger pulp of the non-dominant hand, and the photoplethysmography pulse (PPG) was recorded from the index finger of the same hand. The recorded data was searched sequentially manually to locate the first 30-pulse sequence in which the PPG amplitude of at least six PPG pulses was less than or equal to 60% of the maximum pulse amplitude in the sequence. The primary PPG parameter of interest was PAPPG. For the LDF signal, the pulse amplitude is designated as PALDF, the total LDF for each pulse is designated as LDFTOT,and the LDF pulsatile component is designated as PF. To investigate the relationship between LDF parameters and PAPPG a linear regression analysis of each 30-pulse sequence was done with PAPPG as the independent variable and each of the three LDF parameters individually (PALDF, LDFTOT, and PF) as dependent variables. Results There was a statistically significant direct relationship between PAPPG and all three measures of blood perfusion (p<0.05). Correlation coefficients (R) varied among subjects but within-subject variations versus PAPPG were similar, having mean values that ranged from 0.665 to 0.694. The results also provided evidence in support of a direct relationship between the LDF pulsatility index, defined as the ratio of PF to its mean value., and PAPPG (R=0.779). Conclusions When finger PPG pulse amplitudes are measured in individual subjects there is a moderate-to-strong correlation between the PPG pulse amplitude changes and skin blood perfusion changes. This fact impacts the confidence in using the widely available PPG parameter, peripheral perfusion index, as an indicator of changes in tissue perfusion.However, differences in the PPG pulse amplitude among subjects were less reliable indicators of differences in blood perfusion among subjects. The findings also indicate that a related parameter, the LDF pulsatility index, is also highly correlated with the PPG pulse amplitude and may serve as a useful parameter for future clinical investigations.

Inhibition of nuclear factor-κB activation improves non-nitric oxide-mediated cutaneous microvascular function in reproductive-aged healthy women.

The transcriptional regulator nuclear factor-κB (NF-κB) is a mediator of endothelial dysfunction. Inhibiting NF-κB with salsalate is used to investigate inflammatory mechanisms contributing to accelerated cardiovascular disease risk. However, in the absence of disease, inhibition of NF-κB can impact redox mechanisms, resulting in paradoxically decreased endothelial function. This study aimed to measure microvascular endothelial function during inhibition of the transcriptional regulator NF-κB in reproductive-aged healthy women. In a randomized, single-blind, crossover, placebo-controlled design, nine healthy women were randomly assigned oral salsalate (1,500 mg, twice daily) or placebo treatments for 5 days. Subjects underwent graded perfusion with the endothelium-dependent agonist acetylcholine (ACh, 10-10 to 10-1 M, 33°C) alone and in combination with 15 mM NG-nitro-l-arginine methyl ester [l-NAME; nonselective nitric oxide (NO) synthase inhibitor] through intradermal microdialysis. Laser-Doppler flux was measured over each microdialysis site, and cutaneous vascular conductance (CVC) was calculated as flux divided by mean arterial pressure and normalized to site-specific maximum (CVC%max; 28 mM sodium nitroprusside + 43°C). The l-NAME sensitive component was calculated as the difference between the areas under the dose-response curves. During the placebo and salsalate treatments, the l-NAME sites were reduced compared with the control sites (both P < 0.0001). Across treatments, there was a significant difference between the control and l-NAME sites, where both sites shifted upward following salsalate treatment (both P < 0.0001), whereas the l-NAME-sensitive component was not different (P = 0.94). These data demonstrate that inhibition of the transcriptional regulator NF-κB improves cutaneous microvascular function in reproductive-aged healthy women through non-NO-dependent mechanisms.NEW & NOTEWORTHY The transcription factor nuclear factor-κB (NF-κB) regulates multiple aspects of innate and adaptive immunity by encoding for genes that participate in inflammation and impact endothelial function following NF-κB inhibition with salsalate treatment. Our results show that cutaneous microvascular function is increased through non-nitric oxide (NO)-dependent mechanisms following salsalate treatment in reproductive-aged healthy women.

The Effect of Acute Hyperglycaemia Induced by Oral Glucose Load on Heart Rate Variability and Skin Microvascular Reactivity in Young Adults.

We aimed to elucidate the effects of acute hyperglycaemia, induced by an oral glucose tolerance test (OGTT), on the autonomic nervous system (ANS) and skin microvascular reactivity at the time point of peak plasma glucose concentration (cglc) in 20 young, healthy participants. We assessed their heart rate variability (HRV) as a measure of the ANS activity and the parameters of post-occlusive reactive hyperaemia (PORH) to estimate skin microvascular reactivity as measured by laser Doppler (LD) fluxmetry. The tests were repeated 30 min after a standard OGTT (75 g glucose dissolved in 250 mL water) and, in a separate control experiment, after drinking the same amount of water. Participants had their cglc and serum insulin measured at three consecutive time-points according to the testing protocol. The low-frequency (LF) spectral power, the LF to high-frequency (LF/HF) ratio, and the diastolic blood pressure increased significantly more after water than after OGTT, and there was a trend of the peak LD flux of PORH decreasing more after OGTT than after water. Significant correlations between some PORH and all the HRV parameters and cglc increase after OGTT were found, implying diminished vascular reactivity evoked by hyperglycaemia in healthy subjects with lower glucose tolerance.

Abstract P217: The Role Of Angiotensin-(1-7) In Regulating Vascular Endothelial Function In Postmenopausal Women

Postmenopausal women (PMW) have reduced endothelial function, however, the mechanisms contributing to this decline remain unclear. Angiotensin-(1-7) [Ang-(1-7)] is a vasoactive peptide of the vasodilatory arm of the Renin-Angiotensin system and increasing bioavailable Ang-(1-7) enhances endothelial function in young women (YW) with pre-clinical cardiovascular disease. Circulating Ang-(1-7) decreases with aging and may be impacted by menopause. However, whether increasing bioavailable Ang-(1-7) improves endothelial function in PMW is unknown. The purpose of this study was to test the hypothesis that increasing bioavailable Ang-(1-7) improves endothelial function in PMW. We assessed skin blood flow responses to local heating in 8 healthy YW (27±9 yrs) and 5 healthy PMW (56±4 yrs). Cutaneous vasodilation was measured using laser-Doppler flowmetry directly over microdialysis perfusions of lactated Ringer’s (control) in YW and PMW, and in PMW only, 100 μM Ang-(1-7). Following 45 minutes of perfusion of lactated Ringer’s and Ang-(1-7), skin was heated to 42°C (0.1°C·sec -1 ) until laser-Doppler flux reached a stable plateau, indicative of endothelium-dependent dilation. Following the plateau, 28mM sodium nitroprusside was perfused while skin was heated to 43°C to achieve maximal vasodilation. Cutaneous vascular conductance (CVC) was calculated as laser-Doppler flux/MAP, and normalized to maximal dilation (%CVC max ). Women were normotensive (MAP: YW 86±7 mmHg; PMW 85±9 mmHg) and not obese (BMI: YW 23±3 kg/m 2 ; PMW 23±4 kg/m 2 ). YW had greater dilation during local heating (control: YW 94.54±5.4 vs. PMW 82.59±10.72 %CVC max ; P=0.021). Contrary to our hypothesis, Ang-(1-7) did not enhance heating-induced vasodilation in PMW (80.10±9.43 %CVC max ; P=0.69). These preliminary results suggest that increasing bioavailable Ang-(1-7) may not improve endothelial function in PMW. Additional studies are needed to understand if these findings are due to estrogen deficiency in PMW.

Prefrontal NIRS signal is unaffected by forehead Doppler flux during incremental cycling exercise.

Near-infrared spectroscopy (NIRS) is used to measure tissue concentrations of oxyhemoglobin (O2 Hb) and deoxyhemoglobin (HHb). In the context of exercise, NIRS confers a higher signal-to-noise ratio than other neuroimaging techniques. However, part of the signal may be influenced by thermoregulatory hyperemia in the superficial cutaneous capillaries of the forehead. The degree to which NIRS signals during exercise reflect cerebral or extracerebral hemodynamic changes is a continuing source of controversy. However, the influence of skin blood flow may be attenuated depending on the NIRS technique (e.g., frequency domain machines with maximal optode separation distances >3.5 cm). The purpose of this study was to compare the changes in forehead skin blood flow and cerebral hemoglobin concentration during incremental exercise versus direct vasodilation of the forehead skin induced by gradual local heating. Thirty participants (12 females, 18 males; age: 20.8 ± 3.2 years; body mass index: 23.8 ± 3.7 kg·m-2 ) participated in the study. Forehead skin blood flow was quantified laser Doppler flux and absolute concentrations of cerebral O2 Hb and HHb were measured by NIRS. Local heating significantly increased the Doppler flux signal across time and these changes were significantly correlated with skin temperature. During incremental exercise, skin temperature, Doppler flux, O2 Hband HHb increased however, the only significant change that was consistently correlated with Doppler flux was skin temperature. Therefore, a significant change in forehead skin blood flow may not significantly the NIRS hemoglobin data, depending on the type of NIRS device used.

Cutaneous iontophoresis of adrenergic agonists can mimic in vivo neural effects on vasodilation

Iontophoresis utilizes electrical current to non-invasively deliver agents through the skin. This pilot study sought to determine if iontophoresis could be paired with laser Doppler flowmetry (LDF) to administer adrenergic and cholinergic agonists and measure the subsequent changes in cutaneous blood flow. We hypothesized that iontophoresis administration would produce vascular tone changes consistent with those seen during other delivery methods. We iontophoresed phenylephrine, clonidine, isoproterenol, and acetylcholine to the forearm of one generally healthy 44-year-old female using existing or adapted protocols. We also created half-strength protocols with the goal of establishing dose-response relations. We quantified erythrocyte flux via LDF and beat-by-beat arterial blood pressure via finger photoplethysmography. After baseline, each drug was administered using its half and full-strength protocols, separated by a washout period. Finally, we stimulated maximal blood flow at each iontophoresis site via non-noxious local heating to 43°C. Cutaneous vascular conductance (CVC; laser Doppler flux/ mean arterial pressure) was calculated during baseline and following iontophoresis. We administered vehicle (deionized water) alone as a control, using the same protocols and analysis. The drug responses generally matched our predictions with the exception of phenylephrine, which caused vasodilation. Vehicle administration also caused vasodilation. Full-strength application generally led to a more pronounced change in CVC, suggesting that a dose-response relation might be established with additional data. Applications of certain agents led to a higher CVC than “maximal”, possibly raising the need to alter the method for establishing maximal cutaneous blood flow. Overall, our findings indicate that iontophoresis and LDF can be used to effectively administer adrenergic and cholinergic agonists to non-glabrous skin and affect vascular tone. This non-invasive technique could be used to investigate responses to these agents in those whose cutaneous neural receptor characteristics have been altered as a consequence of disease.

Seven days of statin treatment improves nitric-oxide mediated endothelial-dependent cutaneous microvascular function in women with endometriosis.

Endometriosis is associated with systemic inflammation and increased risk of cardiovascular disease (CVD). Endothelial dysfunction is one of the first manifestations of CVD but is unexplored in women with endometriosis. HMG-CoA-reductase inhibitors (statins) exert potent anti-inflammatory effects, and have been proposed as an adjunctive therapy in women with endometriosis. We hypothesized that microvascular endothelial function would be impaired in otherwise healthy women with endometriosis mediated by reduced nitric oxide (NO)-dependent dilation and that short term statin administration would improve endothelial function. In 8 healthy control (HC: 33±9yr) and 8 women with endometriosis (EN: 34±9yr), laser-Doppler flux (LDF) was measured continuously during graded intradermal microdialysis perfusion of the endothelium-dependent agonist acetylcholine (Ach: 10-10-10-1M) alone and in combination with the NO synthase inhibitor (L-NAME: 0.015M). 6 EN repeated the microdialysis experiment following 7days of oral atorvastatin treatment (10mg). Cutaneous vascular conductance was calculated (CVC=LDF*mmHg-1) and normalized to site-specific maximum (28mM sodium nitroprusside, 43°C). The NO-dependent dilation was calculated as the difference between the areas under the dose response curves. Ach-induced vasodilation was blunted in women with endometriosis (main effect p<0.01), indicating impaired endothelial function. NO-dependent vasodilation was also reduced in women with endometriosis (HC: 217±120.3 AUC vs. EN: 88±97 AUC, p=0.03). Oral atorvastatin improved Ach-induced (main effect p<0.01) and NO-dependent (295±153 AUC; p=0.05) vasodilation in women with endometriosis. Microcirculatory endothelium-dependent vasodilation is impaired in women with endometriosis, mediated in part by reductions in NO. Short-term oral atorvastatin improved endothelium-dependent vasodilation, suggesting that statin therapy may be a viable intervention strategy to mitigate accelerated CVD risk in women with endometriosis.

Effects of systemic and renal intramedullary endothelin-1 receptor blockade on tissue NO and intrarenal hemodynamics in normotensive and hypertensive rats

Endothelin 1 (ET-1) seems essential in salt-dependent hypertension, and activation of ETA receptors causes renal vasoconstriction. However, the response in the renal medulla and the role of tissue NO availability has never been adequately explored in vivo. We examined effects of ETA and ETB receptor blockade (atrasentan and BQ788) on blood pressure (MAP), medullary blood flow (MBF) and medullary tissue NO. Effects of systemic and intramedullary blocker application were compared in anesthetized normotensive ET-1-pretreated Sprague-Dawley rats (S-D), in salt-dependent hypertension (HS/UNX) and in spontaneously hypertensive rats (SHR). Total renal blood flow (RBF) was measured using a Transonic renal artery probe, MBF as laser-Doppler flux, and tissue NO signal using selective electrodes.In normotensive rats ET-1 significantly increased MAP, decreased RBF (−20%) and renal medullary NO. In HS/UNX rats atrasentan decreased MAP and increased medullary NO, earlier and more profoundly with intravenous infusion. In SHR atrasentan decreased MAP, more effectively with intravenous infusion; the increase in tissue NO (∼10%) was similar with both routes; however, only intramedullary atrasentan increased MBF. No consistent responses to BQ788 were seen.We confirmed dominant role of ETA receptors in regulation of blood pressure and renal hemodynamics in normotensive and hypertensive rats and provided novel evidence for the role of ETA in control of intrarenal NO bioavailability in salt-dependent and spontaneous hypertension. Under conditions of activation of the endothelin system ETB stimulation preserved medullary perfusion.

Laser-Doppler microvascular flow of dental pulp in relation to caries progression

Pulpal blood supply can be measured noninvasively and objectively via laser-Doppler fluxmetry. The aim of the study was to measure and compare pulpal laser-Doppler (LD) flux values for permanent non-carious teeth and carious teeth with different degrees of caries progression. Conventional sensitivity tests were also conducted for a comparative analysis to clarify the difference between tooth sensitivity and vitality. Carious lesions were detected and assessed in 15 adult subjects using the International Caries Detection and Assessment System (ICDAS) criteria. Pulpal sensitivity of selected carious teeth (without previous prosthetic restoration or endodontic treatment) and suitable caries-free teeth were then tested with a cold stimulus and subsequently with a weak electric current, whereas their pulpal LD flux was recorded at a separate visit using individually designed silicone probe holders. The LD flux values for teeth with an ICDAS score 6 were significantly lower compared to the values for teeth with an ICDAS score 1 (p < 0.05). Pulpal LD flux values of teeth with active caries were significantly lower than those of teeth with solely inactive carious lesions (p < 0.05). The degree of agreement between sensitivity and vitality testing was fair within both of the pairs: cold test/LDF (κk = 0.232, p = 0.00) and electric test/LDF (κk = 0.354, p = 0.00). Pulpal LD flux measurement proved to be reliable in the pulp vitality assessment of carious and non-carious teeth and might improve the reliability of clinical decisions when used in addition to standard clinical diagnostic protocols.

HYPOTENSIVE EFFECTIVENESS OF THE INTRARENAL MEDULLARY ENDOTHELIN ETA RECEPTOR BLOCKADE IN THE TWO ANIMAL MODELS OF HYPERTENSION

Objective: Endothelin-1 (ET-1) is involved in the pathogenesis of arterial hypertension and chronic kidney disease, however, its mechanism of action is unclear. ET-1 is known to act via ETA and ETB receptors; their activation leads, respectively, to constriction or NO-mediated dilation of the systemic and renal vasculature. ET-1 levels are elevated in experimental and in human hypertension. Here we examined the effects of ETA receptor blockade with atrasentan, administered intravenously (i.v.) or directly to the renal medullary interstitium (i.med.) in two models of rat hypertension. Design and method: In anaesthetized rats with hypertension induced by uninephrectomy followed by high-salt (4% Na w/w) diet (HS/UNX) and in spontaneously hypertensive rats (SHR) we measured mean arterial pressure (MAP), total renal blood flow (RBF, renal artery probe), renal medullary blood flow (MBF, laser-Doppler flux), and renal medullary nitric oxide (NO) signal. Atrasentan doses were: 0.6 mg/kg/h i.v. or 0.12 mg/kg/h i.med. Results: In HS/UNX intravenous atrasentan reduced MAP (142 ± 7 to 117 ± 8 mmHg, p &lt; 0.001) and increased NO signal by 12% (p &lt; 0.05). RBF fell from 25.6 ± 1.7 to 22.3 ± 1.6 ml/min (p &lt; 0.03). Intramedullary atrasentan decreased MAP (127 ± 6 to 115 ± 6, p &lt; 0.04) and increased NO signal by 9.7% (p &lt; 0.03). RBF and MBF did not change significantly. In SHR intravenous atrasentan decreased MAP (171 ± 3 to 152 ± 9 mmHg, p &lt; 0.03) and RBF (6.4 ± 0.5 to 5.0 ± 0.4 ml/min, p &lt; 0.02); NO signal increased by 9.1% (p &lt; 0.01). Intramedullary atrasentan decreased MAP (176 ± 4 to 167 ± 7 mmHg, p &lt; 0.001); NO increased by 10% and MBF by 26% (p &lt; 0.02–0.05). Evidently, in both hypertension models atrasentan always decreased MAP and increased tissue NO. Conclusions: To summarise, in both hypertension models blockade of ETA receptors decreased MAP, reflecting systemic vasodilation. The pressure decrease was followed by a decrease in RBF, probably the result of reduced renal perfusion pressure and imperfect autoregulation. Remarkably, MAP decrease was also seen with intramedullary ETA blockade but then without impairment of renal circulation. Of particular interest was a regular increase in tissue NO: we propose that the blockade of ETA unmasked the action of unbound ET-1 via ETB receptors, leading to NO release.

Real-Time Assessment of Spinal Cord Microperfusion in a Porcine Model of Ischemia/Reperfusion.

Spinal cord injury is a devastating complication of aortic repair. Despite developments for the prevention and treatment of spinal cord injury, its incidence is still considerably high and therefore, influences patient outcome. Microcirculation plays a key role in tissue perfusion and oxygen supply and is often dissociated from macrohemodynamics. Thus, direct evaluation of spinal cord microcirculation is essential for the development of microcirculation-targeted therapies and the evaluation of existing approaches in regard to spinal cord microcirculation. However, most of the methods do not provide real-time assessment of spinal cord microcirculation. The aim of this study is to describe a standardized protocol for real-time spinal cord microcirculatory evaluation using laser-Doppler needle probes directly inserted in the spinal cord. We used a porcine model of ischemia/reperfusion to induce deterioration of the spinal cord microcirculation. In addition, a fluorescent microsphere injection technique was used. Initially, animals were anesthetized and mechanically ventilated. Thereafter, laser-Doppler needle probe insertion was performed, followed by the placement of cerebrospinal fluid drainage. A median sternotomy was performed for exposure of the descending aorta to perform aortic cross-clamping. Ischemia/reperfusion was induced by supra-celiac aortic cross-clamping for a total of 48 min, followed by reperfusion and hemodynamic stabilization. Laser-Doppler Flux was performed in parallel with macrohemodynamic evaluation. In addition, automated cerebrospinal fluid drainage was used to maintain a stable cerebrospinal pressure. After completion of the protocol, animals were sacrificed, and the spinal cord was harvested for histopathological and microsphere analysis. The protocol reveals the feasibility of spinal cord microperfusion measurements using laser-Doppler probes and shows a marked decrease during ischemia as well as recovery after reperfusion. Results showed comparable behavior to fluorescent microsphere evaluation. In conclusion, this new protocol might provide a useful large animal model for future studies using real-time spinal cord microperfusion assessment in ischemia/reperfusion conditions.

Transcutaneous application of the gaseous CO2 for improvement of the microvascular function in patients with diabetic foot ulcers

IntroductionMicrovascular function is impaired in patients with diabetes mellitus (DM) and is involved in numerous DM complications. Several microvascular-supporting interventions have been proposed of which the transcutaneous application of gaseous CO2 (hereinafter CO2 therapy) is one of the most promising. The aim of present study was to determine the effect of repeated CO2 therapies on the cutaneous microvascular function in DM patients with diabetic foot ulcers. MethodologyA total of 42 subjects with at least one chronic diabetic foot ulcer were enrolled in the study. They were divided into the experimental group (21 subjects aged 64.6 ± 11.6 years) that underwent 4-week-long treatment with transcutaneous application of gaseous CO2 (hereinafter CO2 therapies), and the placebo group (21 subjects aged 65.0 ± 10.7 years) that underwent 4-week-long placebo treatment with transcutaneous application of air. Before the first and after the last treatment in both groups, laser Doppler (LD) flux in foot cutaneous microcirculation, heart rate, and arterial blood pressure measurements were carried out during rest and local thermal hyperaemia (LTH) provocation test. ResultsIn the experimental group the following statistically significant changes were observed after the completed treatment 1) increased mean relative powers of LD flux signals during rest in the frequency bands related to NO-independent endothelial (0.07 ± 0.055 vs. 0.048 ± 0.059, p = 0.0058), NO-mediated endothelial (0.154 ± 0.101 vs. 0.113 ± 0.108, p = 0.015), and neurogenic (0.17 ± 0.107 vs. 0.136 ± 0.098, p = 0.018) activity; 2) decreased resting LD flux (35 ± 29 PU vs. 52 ± 56 PU; p = 0.038); and 3) increased peak LD flux as a function of baseline during LTH (482 ± 474%BL vs. 287 ± 262%BL, p = 0.036); there were no statistically significant changes observed in the placebo group. No systemic effects were observed in none of the two groups by means of mean values of heart rate and arterial blood pressure. ConclusionsRepeated CO2 therapies improves the microvasular function in DM patients without any systemic side effects.

Interplay of the adenosine system and NO in control of renal haemodynamics and excretion: Comparison of normoglycaemic and streptozotocin diabetic rats

The adenosine (Ado) system may participate in regulation of kidney function in diabetes mellitus (DM), therefore we explored its role and interrelation with NO in the control of renal circulation and excretion in normoglycemic (NG) and streptozotocin-diabetic (DM) rats. Effects of theophylline (Theo), a non-selective Ado receptor antagonist, were examined in anaesthetized NG or in streptozotocin induced diabetic (DM) rats, untreated or after blockade of NO synthesis with l-NAME. We measured arterial blood pressure (MABP), whole kidney blood flow and renal regional flows: cortical and outer- and inner-medullary (IMBF), determined as laser-Doppler fluxes. Renal excretion of water, total solutes and sodium and in situ renal tissue NO signal (selective electrodes) were also determined. Theo experiments disclosed minor baseline vasoconstrictor and vasodilator tone in the kidney of NG and DM rats, respectively. NO blockade increased baseline MABP and decreased renal haemodynamics, similarly in NG and DM rats, indicating comparable vasodilator influence of NO in the two groups. Unexpectedly, in all rats with intact NO synthesis, Ado receptor blockade increased kidney tissue NO. In NO-deficient NG and DM rats, Ado receptor blockade induced comparable renal vasodilatation, suggesting similar vasoconstrictor influence of the Ado system. However, DM rats showed an unexplained association of decreased MABP and IMBF and increased NO signal. Higher baseline renal excretion in DM rats indicated inhibition of renal tubular reabsorption due to the prevalence of natriuretic A2 over antinatriuretic A1 receptors. In conclusion, the experiments provided new insights in functional interrelation of adenosine and NO in normoglycaemia and streptozotocin-diabetes.

Normobaric hypoxia does not alter the critical environmental limits for thermal balance during exercise‐heat stress

What is the central question of this study? Hypoxia reportedly does not impair thermoregulation during exercise in compensable heat stress conditions: does it have an impact on maximal heat dissipation and therefore the critical environmental limit for the physiological compensability of core temperature? What is the main finding and its importance? Although skin blood flow was higher in hypoxia, no differences in sweat rates or the critical environmental limit for the physiological compensability of core temperature - an indicator of maximal heat loss - were found compared to exercise in normoxia, indicating no influence of normobaric hypoxia on thermoregulatory capacity in warm conditions. Altered control of skin blood flow (SkBF) in hypoxia does not impair thermoregulation during exercise in compensable conditions, but its impact on maximal heat dissipation is unknown. This study therefore sought to determine whether maximum heat loss is altered by hypoxia during exercise in warm conditions. On separate days, eight males exercised for 90min at a fixed heat production of ∼500W in normoxia (NORM) or normobaric hypoxia (HYP, = 0.13) in a 34°C environment. Ambient vapour pressure was maintained at 2.13kPa for 45min, after which it was raised 0.11kPa every 7.5min. The critical ambient vapour pressure at which oesophageal temperature inflected upward (Pcrit ) indicated that maximum heat dissipation had been reached. Neither local sweat rates on the upper arm, back and forehead (average NORM: 1.46 (0.15) vs. HYP: 1.41 (0.16) mgcm-2 min-1 ; P=0.59) nor whole-body sweat losses (NORM: 1029 (137) g vs. HYP: 1025 (150) g; P=0.95) were different between trials. Laser-Doppler flux values (LDF; arbitrary units), an index of SkBF, were not different between NORM and HYP on the forearm (P=0.23) or back (P=0.73); however, when normalized as a percentage of maximum, LDF values tended to be higher in HYP compared to NORM at the forearm (condition effect, P=0.05) but not back (P=0.19). Despite potentially greater SkBF in hypoxia, there was no difference in Pcrit between conditions (NORM: 3.67 (0.35) kPa; HYP: 3.46 (0.39) kPa; P=0.22). These findings suggest that hypoxia does not independently alter thermoregulatory capacity during exercise in warm conditions.

A Physiological Validation of Spectral Analysis for Skin Flowmotion

In a series of studies, our laboratory has utilized microneurography coupled with laser Doppler flowmetry to examine sympathetic neural control of the cutaneous microvasculature during thermoregulatory stimuli. However, these techniques have limited clinical utility. Spectral analysis of the low frequency periodic oscillations in laser Doppler flowmetry‐derived red blood cell flux (i.e., skin flowmotion) is a noninvasive technique that has been used to quantify cutaneous microvascular control mechanisms (e.g., endothelial, neurogenic, myogenic). To date, conclusions regarding the mechanistic regulation of cutaneous microvascular function derived from spectral analysis have been primarily limited to basal conditions. Thus, the aim here was to examine the validity of spectral analysis to quantify sympathetic neural control of skin blood flow during a dynamic physiological stimulus (local skin heating to 42°C). We hypothesized that pharmacological blockade of sensory nerves would reduce the spectral power of the neurogenic frequency band during local heating. Eleven young healthy adults (7 women; 22±3 yrs, 25±2 kg/m2) participated. Laser Doppler flux was measured continuously during baseline (33°C) and a standardized local heating protocol (42°C) at a non‐treated control site (con) and a site treated with topical lidocaine (lido; 2.5% prilocaine+2.5% lidocaine) to pharmacologically block sensory nerves. Topical lido was applied for a minimum of 45 minutes prior to testing. Cutaneous vascular conductance was calculated and expressed as a percentage of maximum (local heat of 43°C). A raw Fourier transform was used to calculate spectral power around the 0.0095–0.02 Hz, 0.02–0.05 Hz, and 0.05–0.15 Hz frequency intervals, considered to correspond to endothelial, neurogenic, and myogenic activity, respectively. The initial peak of the local heating response was reduced by lido, (con: 62±2% vs. lido: 25±5%, p&lt;0.01), validating adequate sensory nerve blockade. However, there was no difference between sites during baseline (con: 12±1% vs. lido: 9±2%, p=0.07) or the local heating‐induced plateau (con: 95±3% vs. lido: 87±7%, p=0.20). There was no effect of lido on spectral power of any of the frequency intervals at baseline or during local heating. Total spectral power (i.e., the sum of the spectral power for all frequency bands) was significantly related to conductance in both sites at baseline (con: R2=0.69, p&lt;0.01; lido: R2=0.72, p&lt;0.01) and during local heating (con: R2=0.22, p&lt;0.02; lido: R2=0.71, p&lt;0.01). In conclusion, the physiological increase in cutaneous vascular conductance during local heating was mirrored by an increase in total spectral power. Although blockade of sensory nerves reduced the increase in conductance during the initial peak of the local heating response, it did not affect the neurogenic frequency interval derived from spectral analysis. These preliminary data suggest that skin flowmotion may not be a valid approach to evaluate neural control of cutaneous microvascular function during dynamic conditions.Support or Funding InformationAmerican Heart Association Grant #18UFEL33900164; College of Health and Human Development Summer 2019 Undergraduate Research Award; Noll Endowment for Undergraduate Research

Cold Renal Perfusion During Simulation of Juxtarenal Aortic Aneurysm Repair Reduces Systemic Oxidative Stress and Sigmoid Damage in Rats

Juxtarenal aortic surgery induces renal ischaemia reperfusion, which contributes to systemic inflammatory tissue injury and remote organ damage. Renal cooling during suprarenal cross clamping has been shown to reduce renal damage. It is hypothesised that renal cooling during suprarenal cross clamping also has systemic effects and could decrease damage to other organs, like the sigmoid colon. Open juxtarenal aortic aneurysm repair was simulated in 28 male Wistar rats with suprarenal cross clamping for 45 min, followed by 20 min of infrarenal aortic clamping. Four groups were created: sham, no, warm (37 °C saline), and cold (4 °C saline) renal perfusion during suprarenal cross clamping. Primary outcomes were renal damage and sigmoid damage. To assess renal damage, procedure completion serum creatinine rises were measured. Peri-operative microcirculatory flow ratios were determined in the sigmoid using laser Doppler flux. Semi-quantitative immunofluorescence microscopy was used to measure alterations in systemic inflammation parameters, including reactive oxygen species (ROS) production in circulating leukocytes and leukocyte infiltration in the sigmoid. Sigmoid damage was assessed using digestive enzyme (intestinal fatty acid binding protein - I-FABP) leakage, a marker of intestinal integrity. Suprarenal cross clamping caused deterioration of all systemic parameters. Only cold renal perfusion protected against serum creatinine rise: 0.45 mg/dL without renal perfusion, 0.33 mg/dL, and 0.14 mg/dL (p = .009) with warm and cold perfusion, respectively. Microcirculation in the sigmoid was attenuated with warm (p = .002) and cold renal perfusion (p = .002). A smaller increase of ROS production (p = .034) was seen only after cold perfusion, while leukocyte infiltration in the sigmoid colon decreased after warm (p = .006) and cold perfusion (p = .018). Finally, digestive enzyme leakage increased more without (1.5AU) than with warm (1.3AU; p = .007) and cold renal perfusion (1.2AU; p = .002). Renal ischaemia/reperfusion injury after suprarenal cross clamping decreased microcirculatory flow, increased systemic ROS production, leukocyte infiltration, and I-FABP leakage in the sigmoid colon. Cold renal perfusion was superior to warm perfusion and reduced renal damage and had beneficial systemic effects, reducing sigmoid damage in this experimental study.

Cold Renal Perfusion During Simulation of Juxtarenal Aortic Aneurysm Repair Reduces Systemic Oxidative Stress and Sigmoid Damage in Rats

Juxtarenal aortic surgery induces renal ischaemia reperfusion, which contributes to systemic inflammatory tissue injury and remote organ damage. Renal cooling during suprarenal cross clamping has been shown to reduce renal damage. It is hypothesised that renal cooling during suprarenal cross clamping also has systemic effects and could decrease damage to other organs, like the sigmoid colon. Open juxtarenal aortic aneurysm repair was simulated in 28 male Wistar rats with suprarenal cross clamping for 45min, followed by 20min of infrarenal aortic clamping. Four groups were created: sham, no, warm (37°C saline), and cold (4°C saline) renal perfusion during suprarenal cross clamping. Primary outcomes were renal damage and sigmoid damage. To assess renal damage, procedure completion serum creatinine rises were measured. Peri-operative microcirculatory flow ratios were determined in the sigmoid using laser Doppler flux. Semi-quantitative immunofluorescence microscopy was used to measure alterations in systemic inflammation parameters, including reactive oxygen species (ROS) production in circulating leukocytes and leukocyte infiltration in the sigmoid. Sigmoid damage was assessed using digestive enzyme (intestinal fatty acid binding protein - I-FABP) leakage, a marker of intestinal integrity. Suprarenal cross clamping caused deterioration of all systemic parameters. Only cold renal perfusion protected against serum creatinine rise: 0.45mg/dL without renal perfusion, 0.33mg/dL, and 0.14mg/dL (p=.009) with warm and cold perfusion, respectively. Microcirculation in the sigmoid was attenuated with warm (p=.002) and cold renal perfusion (p=.002). A smaller increase of ROS production (p=.034) was seen only after cold perfusion, while leukocyte infiltration in the sigmoid colon decreased after warm (p=.006) and cold perfusion (p=.018). Finally, digestive enzyme leakage increased more without (1.5AU) than with warm (1.3AU; p=.007) and cold renal perfusion (1.2AU; p=.002). Renal ischaemia/reperfusion injury after suprarenal cross clamping decreased microcirculatory flow, increased systemic ROS production, leukocyte infiltration, and I-FABP leakage in the sigmoid colon. Cold renal perfusion was superior to warm perfusion and reduced renal damage and had beneficial systemic effects, reducing sigmoid damage in this experimental study.

Oral L-Tyrosine Supplementation Improves Core Temperature Maintenance in Older Adults.

During cold exposure, an increase in sympathetic nerve activity evokes vasoconstriction (VC) of cutaneous vessels to minimize heat loss. In older adults, this reflex VC response is impaired thereby increasing their susceptibility to excess heat loss and hypothermia. Because L-tyrosine, the amino acid substrate necessary for catecholamine production, has been shown to augment reflex VC in age skin, we hypothesize that oral ingestion of L-tyrosine will attenuate the decline in core temperature (Tc) during whole-body cooling in older adults. In a randomized, double-blind design, nine young (25 ± 3 yr) and nine older (72 ± 8 yr) participants ingested either 150 mg·kg of L-tyrosine or placebo before commencing 90 min of whole-body cooling to decrease skin temperature to approximately 29.5°C. Esophageal temperature and forearm laser Doppler flux (LDF) were measured continuously throughout the protocol to provide an index of Tc and skin blood flow, respectively. The change in esophageal temperature (ΔTES) was the difference in temperature at the end of cooling subtracted from baseline. Cutaneous vascular conductance (CVC) was calculated as CVC = LDF/mean arterial pressure and expressed as a percent change from baseline (%ΔCVCBASELINE). Oral tyrosine ingestion augmented the cutaneous VC response to cooling in older adults (placebo, 14.4 ± 2.0; tyrosine, 32.7% ± 1.7% ΔCVCBASELINE; P < 0.05). Additionally, tyrosine improved Tc maintenance throughout cooling in older adults (placebo, -0.29 ± 0.07; tyrosine, -0.07 ± 0.07 ΔTES; P < 0.05). Both the cutaneous VC and Tc during cooling were similar between young and older adults supplemented with tyrosine (P > 0.05). These results indicate that L-tyrosine supplementation improves Tc maintenance in response to acute cold exposure in an older population.

The acute effects of different levels of intermittent negative pressure on peripheral circulation in patients with peripheral artery disease.

Intermittent negative pressure (INP) applied to the lower leg induces acute increase in arterial and skin blood flow. The aim of this study was to identify the optimal level of INP to increase blood flow in patients with lower extremity peripheral artery disease (PAD). We investigated the acute effects of different levels of INP in 16 subjects (7 women and 9 men, mean (SD) age 71(8) years) diagnosed with PAD. During application of INP in a pressure chamber sealed below the knee, arterial blood flow was continuously recorded in the dorsalis pedis artery or tibialis posterior artery (ultrasound Doppler), and skin blood flow was continuously recorded at the pulp of the first toe (laser Doppler). Different pressure levels (0, −10, −20, −40, and −60 mmHg) were tested in randomized order. Maximal arterial blood flow relative to baseline (median [25th, 75th percentiles]) was: 0 mmHg; 1.08 (1.02, 1.13), −10 mmHg; 1.11 (1.07, 1.17), −20 mmHg; 1.18 (1.11, 1.32), −40 mmHg; 1.39 (1.27, 1.91) and −60 mmHg; 1.48 (1.37, 1.78). Maximal laser Doppler flux (LDF) relative to baseline was: 0 mmHg; 1.06 (1.02, 1.12), −10 mmHg; 1.08 (1.05, 1.16) −20 mmHg; 1.12 (1.06, 1.27), −40 mmHg; 1.24 (1.14, 1.50) and −60 mmHg; 1.35 (1.10, 1.70). There were significantly higher maximal arterial blood flow and maximal LDF at −40 mmHg compared with −10 mmHg (P = 0.001 and P = 0.025, respectively). There were no significant differences in maximal arterial blood flow and maximal LDF between 0 and −10 mmHg (both P = 1.0), or between −40 and −60 mmHg (both P = 1.0). INP of −40 mmHg was the lowest negative pressure level that increased blood flow.

Arginine and tetrahydrobiopterin supplementation in rats with salt-induced blood pressure increase: minor hypotensive effect but improvement of renal haemodynamics.

High salt (HS) intake can lead to hypertension, probably the result of the predominance of vasoconstrictor reactive oxygen species over vasodilator nitric oxide (NO). We aimed to examine if the supposed NO deficiency and the resultant blood pressure increase could be corrected by supplementation of L-arginine, the substrate, and tetrahydrobiopterin (BH4), a co-factor of NO synthases. Wistar rats without known genetic background of salt sensitivity were exposed to HS diet (4%Na) for 10 or 26 days, without or with supplementation with oral L-arginine, 1.4 mg/kg b.w. daily, alone or together with intraperitoneal BH4, 10 mg/kg daily. Systolic blood pressure (SBP, tail-cuff method) was measured repeatedly and found to increase ~40 mmHg after 26 days; L-arginine and BH4 did not significantly attenuate this increase. At the end of chronic studies, in anaesthetized rats the diet- and treatment-induced changes in renal haemodynamics were assessed. HS diet selectively decreased (-30%, P < 0.03) the inner medullary blood flow (IMBF, laser-Doppler flux) without changing total or cortical renal perfusion. Arginine supplementation tended to raise all renal circulatory parameters, and distinctly increased IMBF, to 61% above the HS diet level (P < 0.05). In conclusion, unlike in confirmed genetically determined salt-dependent hypertension, L-arginine and BH4 supplementation failed to attenuate the SBP increase observed after exposure to HS diet. On the other hand, arginine increased total and regional renal perfusion, especially IMBF. This suggests that the delivery of arginine increased intrarenal NO synthesis, an action of renoprotective potential which presumably countered the harmful influence of the local tissue oxidative stress.