Search

Idiopathic inflammatory myopathies (IIM) are characterized by muscle inflammation and weakness, myositis-specific autoantibodies (MSAs), and extramuscular organ damage. The role of neutrophil dysregulation and neutrophil extracellular traps (NETs) in IIM is unclear. We assessed whether pathogenic neutrophil subsets (low-density granulocytes [LDGs]) and NETs were elevated in IIM, associated with clinical presentation and MSAs, and their effect on skeletal myoblasts and myotubes. Circulating NETs and LDGs were quantified and correlated with clinical measures. Specific MSAs were tested for their ability to induce NETs. NETs and neutrophil gene expression were measured in IIM biopsies. Whether NETs damage skeletal myoblasts and myotubes was tested. Circulating LDGs and NETs were increased in IIM. IIM LDGs had an enhanced ability to form NETs. LDGs and NETs correlated with IIM disease activity and muscle damage. The serum MSA anti-MDA5 correlated with circulating and tissue NETs and directly enhanced NET formation. An enhanced neutrophil gene signature was present in IIM muscle and associated with muscle injury and tissue IFN gene signatures. IIM NETs decreased the viability of myotubes in a citrullinated histone-dependent manner. Dysregulated neutrophil pathways may play pathogenic roles in IIM through their ability to directly injure muscle cells and other affected tissues.

BACKGROUND Patient journals have been used as valuable resources in clinical studies. However, the full potential value of such journals can be undermined by inefficiencies and ambiguities associated with handwritten patient reports. The increasing number of mobile phones and mobile-based health care approaches presents an opportunity to improve communications from patients to clinicians and clinical researchers through the use of digital patient journals. OBJECTIVE The objective of this project was to develop a smartphone-based platform that would enable patients to record events and symptoms on the same timeline as clinical data collected by wearable sensors. METHODS This platform consists of two major components: a smartphone for patients to record their journals and wireless sensors for clinical data collection. The clinical data and patient records are then exported to a clinical researcher interface, and the data and journal are processed and combined into a single time-series graph for analysis. This paper gives a block diagram of the platform’s principal components and compares its features to those of other methods but does not explicitly discuss the process of design or development of the system. RESULTS As a proof of concept, body temperature data were obtained in a 4-hour span from a 22-year-old male, during which the subject simultaneously recorded relevant activities and events using the iPhone platform. After export to a clinical researcher’s desktop, the digital records and temperature data were processed and fused into a single time-series graph. The events were filtered based on specific keywords to facilitate data analysis. CONCLUSIONS We have developed a user-friendly patient journal platform, based on widely available smartphone technology, that gives clinicians and researchers a simple method to track and analyze patient activities and record the activities on a shared timeline with clinical data from wearable devices.

Summary Background Well-tolerated and effective treatments are needed for chronic leg ulcers in sickle cell anaemia. Topical sodium nitrite, a known nitric oxide donor, enhances blood flow in ulcers and has known bacteriostatic effects. We aimed to assess the safety, tolerability, and pharmacokinetics of topical sodium nitrite in patients with sickle cell disease and chronic leg ulcers. Methods We enrolled adult patients from an ambulatory clinic at the National Institutes of Health (Bethesda, MD, USA) with sickle cell anaemia with leg ulcers (with a surface area of 2·5–100 cm 2 ) persisting for at least 4 weeks into a safety and tolerability phase 1 dose-escalation trial of topical sodium nitrite. Increasing concentrations of sodium nitrite cream were applied twice weekly for 4 weeks to one ulcer per patient at five dose levels (0·5%, 1%, 1·5%, 1·8%, and 2%). The primary endpoints were safety and tolerability, with secondary endpoints of pharmacokinetics, blood flow, and wound healing. Pain relief was analysed post hoc. Endpoints were analysed over time for the whole study population and according to dose level. This study is registered with ClinicalTrials.gov, number NCT01316796. Findings Between April 4, 2011, and March 19, 2013, we enrolled 18 adult patients with sickle cell anaemia and leg ulcers into our trial. We assigned three patients into each cohort, and each cohort was treated with a different concentration of sodium nitrite cream (cohort 1: 0·5%, cohort 2: 1·0%, cohort 3: 1·5%, and cohort 4: 2·0%). Patients were not enrolled into the next cohort dose until we were able to establish that no dose-limiting toxicities were observed. An additional six patients were enrolled to cohort 3a: 1·8%, after two patients in cohort 4 had asymptomatic drops in diastolic blood pressure. No grade 3–4 adverse events were observed, and there were no serious adverse events or dose-limiting side-effects. Pharmacokinetic analysis showed that systemic absorption of sodium nitrite was very low. Application of topical sodium nitrite was associated with a significant increase in peri-wound cutaneous blood flow measured by laser speckle contrast imaging (p=0·0002), corroborated by increased peri-wound skin temperature by infrared thermography (p=0·0119). We recorded a dose-dependent decrease in leg ulcer size (p=0·0012) and pain (p Interpretation Our results indicate that topical sodium nitrite 2% cream is suitable for additional clinical trials in adults with sickle cell anaemia to promote healing of leg ulcers. Funding National Heart, Lung and Blood Institute Division of Intramural Research (National Institutes of Health).

The present study was aimed to in vivo assess the blood flow oscillatory patterns in rat pial microvessels during 30 min bilateral common carotid artery occlusion (BCCAO) and 60 min reperfusion by laser speckle imaging. Pial microcirculation was visualized by fluorescence microscopy. The blood flow oscillations of single microvessels were recorded by laser speckle imaging; spectral analysis was performed by Wavelet transform. Under baseline conditions, arterioles and venules were characterized by blood flow oscillations in the frequency ranges 0.005-0.0095 Hz, 0.0095-0.021 Hz, 0.021-0.052 Hz, 0.052-0.150 Hz and 0.150-0.500 Hz. Arterioles showed oscillations with the highest spectral density when compared with venules. Moreover, the frequency components in the ranges 0.052-0.150 Hz and 0.150-0.500 were predominant in the arteriolar total power spectrum; while, the frequency component in the range 0.150-0.500 Hz showed the highest spectral density in venules. After 30 min BCCAO, the arteriolar spectral density decreased compared to baseline; moreover, the arteriolar frequency component in the range 0.052-0.150 Hz significantly decreased in percent spectral density, while the frequency component in the range 0.150-0.500 Hz significantly increased in percent spectral density. However, an increase in arteriolar spectral density was detected at 60 min reperfusion compared to BCCAO values; consequently, an increase in percent spectral density of the frequency component in the range 0.052-0.150 Hz was observed, while the percent spectral density of the frequency component in the range 0.150-0.500 Hz significantly decreased. The remaining frequency components did not significantly change during hypoperfusion and reperfusion. The changes in blood flow during hypoperfusion/reperfusion caused tissue damage in the cortex and striatum of all animals. In conclusion, our data demonstrate that the frequency component in the range 0.052-0.150 Hz, related to myogenic activity, was significantly impaired by hypoperfusion and reperfusion, affecting cerebral blood flow distribution and causing tissue damage.

Resistance to chemotherapy is the primary cause of treatment failure in over 90% of cancer patients in the clinic. Research in nanotechnology-based therapeutic alternatives has helped provide innovative and promising strategies to overcome multidrug resistance (MDR). By targeting CD44-overexpressing MDR cancer cells, we have developed in a single-step a self-assembled, self-targetable, therapeutic semiconducting single-walled carbon nanotube (sSWCNT) drug delivery system that can deliver chemotherapeutic agents to both drug-sensitive OVCAR8 and resistant OVCAR8/ADR cancer cells. The novel nanoformula with a cholanic acid-derivatized hyaluronic acid (CAHA) biopolymer wrapped around a sSWCNT and loaded with doxorubicin (DOX), CAHA-sSWCNT-DOX, is much more effective in killing drug-resistant cancer cells compared to the free DOX and phospholipid PEG (PL-PEG)-modified sSWCNT formula, PEG-sSWCNT-DOX. The CAHA-sSWCNT-DOX affects the viscoelastic property more than free DOX and PL-PEG-sSWCNT-DOX, which in turn allows more drug molecules to be internalized. Intravenous injection of CAHA-sSWCNT-DOX (12 mg/kg DOX equivalent) followed by 808 nm laser irradiation (1 W/cm(2), 90 s) led to complete tumor eradication in a subcutaneous OVCAR8/ADR drug-resistant xenograft model, while free DOX alone failed to delay tumor growth. Our newly developed CAHA-sSWCNT-DOX nanoformula, which delivers therapeutics and acts as a sensitizer to influence drug uptake and induce apoptosis with minimal resistance factor, provides a novel effective means of counteracting the phenomenon of multidrug resistance.

Background Patient journals have been used as valuable resources in clinical studies. However, the full potential value of such journals can be undermined by inefficiencies and ambiguities associated with handwritten patient reports. The increasing number of mobile phones and mobile-based health care approaches presents an opportunity to improve communications from patients to clinicians and clinical researchers through the use of digital patient journals. Objective The objective of this project was to develop a smartphone-based platform that would enable patients to record events and symptoms on the same timeline as clinical data collected by wearable sensors. Methods This platform consists of two major components: a smartphone for patients to record their journals and wireless sensors for clinical data collection. The clinical data and patient records are then exported to a clinical researcher interface, and the data and journal are processed and combined into a single time-series graph for analysis. This paper gives a block diagram of the platform's principal components and compares its features to those of other methods but does not explicitly discuss the process of design or development of the system. Results As a proof of concept, body temperature data were obtained in a 4-hour span from a 22-year-old male, during which the subject simultaneously recorded relevant activities and events using the iPhone platform. After export to a clinical researcher's desktop, the digital records and temperature data were processed and fused into a single time-series graph. The events were filtered based on specific keywords to facilitate data analysis. Conclusions We have developed a user-friendly patient journal platform, based on widely available smartphone technology, that gives clinicians and researchers a simple method to track and analyze patient activities and record the activities on a shared timeline with clinical data from wearable devices.

Chronic leg ulcers are frequent and debilitating complications of sickle cell anemia. Inadequate blood supply has been postulated to be an important factor in their occurrence and delayed healing. Little is known about their microcirculatory and histopathological changes. We evaluated the microcirculation of lower extremity ulcers with laser speckle contrast imaging and infrared thermography and obtained clinical and laboratory characteristics in 18 adults with sickle cell anemia and chronic leg ulcers. Skin biopsies were obtained in four subjects. Subjects had markers of severe disease, anemia, high degree of hemolysis, inflammation, and thrombophilia. The highest blood flow was present in the ulcer bed, progressively less in the immediate periwound area, and an unaffected control skin area in the same extremity. Microscopic examination showed evidence of venostasis, inflammation, and vasculopathy. Blood vessels were increased in number, had activated endothelium and evidence of thrombosis/recanalization. High blood flow may be due to chronic inflammation, cutaneous vasodilatation, venostasis, and in situ thrombosis. These changes in skin microcirculation are similar to chronic venous ulcers in the non-sickle cell disease (SCD) population, thus suggesting that leg ulcers may be another end-organ complication with endothelial dysfunction that appears in patients with SCD at a younger age and with higher frequency than in the general population.

Vascular dysfunction is an important pathophysiologic manifestation of sickle cell disease (SCD), a condition that increases risk of pulmonary hypertension and stroke. We hypothesized that infrared (IR) imaging would detect changes in cutaneous blood flow reflective of vascular function. We performed IR imaging and conventional strain gauge plethysmography in twenty-five adults with SCD at baseline and during intra-arterial infusions of an endothelium-dependent vasodilator acetylcholine (ACh), an endothelium-independent vasodilator sodium nitroprusside (SNP), and a NOS inhibitor L-NMMA. Skin temperature measured by IR imaging increased in a dose-dependent manner to graded infusions of ACh (+1.1° C, p < 0.0001) and SNP (+0.9° C, p < 0.0001), and correlated with dose-dependent increases in forearm blood flow (ACh: +19.9 mL/min/100mL, p < 0.0001; rs = 0.57, p = 0.003; SNP: +8.6 mL/min/100mL, p < 0.0001; r = 0.70, p = 0.0002). Although IR measurement of skin temperature accurately reflected agonist-induced increases in blood flow, it was less sensitive to decreases in blood flow caused by NOS inhibition. Baseline forearm skin temperature measured by IR imaging correlated significantly with baseline forearm blood flow (31.8±0.2° C, 6.0±0.4 mL/min/100mL; r = 0.58, p = 0.003), and appeared to represent a novel biomarker of vascular function. It predicted a blunted blood flow response to SNP (r = −0.61, p = 0.002), and was independently associated with a marker of pulmonary artery pressure, as well as hemoglobin level, diastolic blood pressure, homocysteine, and cholesterol (R2 = 0.84, p < 0.0001 for the model). IR imaging of agonist-stimulated cutaneous blood flow represents a less cumbersome alternative to plethysmography methodology. Measurement of baseline skin temperature by IR imaging may be a useful new marker of vascular risk in adults with SCD.

We show that a mobile phone can serve as an accurate monitor for several physiological variables, based on its ability to record and analyze the varying color signals of a fingertip placed in contact with its optical sensor. We confirm the accuracy of measurements of breathing rate, cardiac R-R intervals, and blood oxygen saturation, by comparisons to standard methods for making such measurements (respiration belts, ECGs, and pulse-oximeters, respectively). Measurement of respiratory rate uses a previously reported algorithm developed for use with a pulse-oximeter, based on amplitude and frequency modulation sequences within the light signal. We note that this technology can also be used with recently developed algorithms for detection of atrial fibrillation or blood loss.

In this work we demonstrate that functional infrared imaging is capable of detecting low frequency temperature fluctuations in intact human skin and revealing spatial, temporal, spectral, and time-frequency based differences among three tissue classes: microvasculature, large sub-cutaneous veins, and the remaining surrounding tissue of the forearm. We found that large veins have stronger contractility in the range of 0.005-0.06 Hz compared to the other two tissue classes. Wavelet phase coherence and power spectrum correlation analysis show that microvasculature and skin areas without vessels visible by IR have high phase coherence in the lowest three frequency ranges (0.005-0.0095 Hz, 0.0095-0.02 Hz, and 0.02-0.06 Hz), whereas large veins oscillate independently.

Chronotherapeutics involve the administration of treatments according to circadian rhythms. Circadian timing of anti-cancer medications has been shown to improve treatment tolerability up to fivefold and double efficacy in experimental and clinical studies. However, the physiological and the molecular components of the circadian timing system (CTS), as well as gender, critically affect the success of a standardized chronotherapeutic schedule. In addition, a wrongly timed therapy or an excessive drug dose disrupts the CTS. Therefore, a non-invasive approach to accurately detect and monitor circadian rhythms is needed for a dynamic assessment of the CTS in order to personalize chronomodulated drug delivery schedule in cancer patients. Since core body temperature is a robust circadian biomarker, we recorded temperature at multiple locations on the skin of the upper chest and back of controls and cancer patients continuously. Variability in the circadian phase existed among patch locations in individual subjects over the course of 2–6 days, demonstrating the need to monitor multiple skin temperature locations to determine the precise circadian phase. Additionally, we observed that locations identified by infrared imaging as relatively cool had the largest 24 h temperature variations. Disruptions in skin temperature rhythms during treatment were found, pointing to the need to continually assess circadian timing and personalize chronotherapeutic schedules.

Background: Human obesity is associated with increased heat production; however, subcutaneous adipose tissue provides an insulating layer that impedes heat loss. To maintain normothermia, therefore, obese individuals must increase their heat dissipation. Objective: The objective was to test the hypothesis that temperature in a heat-dissipating region of the hand is elevated in obese adults. Design: Obese [body mass index (in kg/m 2 ) 30] and normalweight (NW; body mass index = 18–25) adults were studied under thermoneutral conditions at rest. Core body temperature was measured by using ingested telemetric capsules. The temperatures of the third fingernail bed of the right hand and of abdominal skin from an area 1.5 cm inferior to the umbilicus were determined by using infrared thermography. Abdominal skin temperatures were also measured via adhesive thermistors that were placed over a prominent skin-surface blood vessel and over an adjacent nonvessel location. The groups were compared by analysis of covariance with age, sex, race, and room temperature as covariates. Results: Core temperature did not differ significantly between the 23 obese and 13 NW participants (P = 0.74). However, infrared thermography–measured fingernail-bed temperature was significantly higher in obese subjects than in NW subjects (33.9 6 0.7C compared with 28.6 6 0.9C; P , 0.001). Conversely, infrared thermography–measured abdominal skin temperature was significantly lower in obese subjects than in NW subjects (31.8 6 0.2C compared with 32.8 6 0.3C; P = 0.02). Nonvessel abdominal skin temperatures measured by thermistors were also lower in obese subjects (P = 0.04). Conclusions: Greater subcutaneous abdominal adipose tissue in obese adults may provide a significant insulating layer that blunts abdominal heat transfer. Augmented heat release from the hands may offset heat retention in areas of the body with greater adiposity, thereby helping to maintain normothermia in obesity. This trial was registered at clinicaltrials.gov as NCT00266500. Am J Clin Nutr 2009;90:1124–31.

Background Currently visual and tactile clues such as color, mottling, and tissue turgor are used in the operating room for subjective assessments of organ ischemia. Studies have demonstrated that infrared (IR) imaging is a reliable tool to identify perfusion of brain tumors and kidneys during human surgery. Intraoperative IR imaging has the potential for more objective real-time detection and quantitative assessment of organ viability including early ischemia. We hypothesize, by detecting variations of the IR signal, we can assess the degree to which renal surface temperature reflects underlying renal ischemia. To address this hypothesis, IR imaging-derived temperature fluctuations were evaluated during laparotomy in a porcine model ( n = 15). These temperature profiles then underwent spectral (frequency) analysis to assess their relationship to well-described oscillations of the microcirculation. Materials and methods An IR camera was positioned 30–60 cm above the exposed kidneys. Images (3–5 μm wavelength) were collected (1.0/s) at baseline, during warm renal ischemia, and during reperfusion. Dominant frequency ( DF ) of the tissue temperature fluctuations were determined by a Fourier transformation (spectral) analysis. Results IR images immediately showed which segments of the kidney were ischemic. DF at ∼0.008 Hz that corresponds to blood flow oscillations was observed in thermal profiles. The oscillations were diminished or disappeared after 25 min of warm ischemia and were recovered with reperfusion in a time-dependent fashion. Oscillations were attenuated substantially in ischemic segments, but not in perfused segments of the kidney. Conclusions The described oscillations can be measured noninvasively using IR imaging in the operating room, as represented by the DF , and may be an early marker of critical renal ischemia.

A high-resolution infrared (IR) camera was used to assess rhythmicity in localized renal blood flow, including the extent of regions containing nephrons with spontaneous oscillations in their individual blood flow. The IR imaging was able to follow changes in rat renal perfusion during baseline conditions, during occlusion of the main renal artery and during the administration of either saline or papaverine. Concurrent recordings were made of tubular pressure in superficial nephrons. Spontaneous vascular oscillations centred around 0.02–0.05 Hz and approximately 0.01 Hz could be detected reproducibly by IR imaging. Their spectral characteristics and their response to papaverine were in line with tubular pressure measurements. The intensity of and synchrony between thermal signals from different local areas of the kidney may allow, after surgical exposure, non-invasive imaging of functional clusters involved in renal cortical blood flow. Through visualization of the spatial extent of thermal oscillations, IR imaging holds promise in assessing kidney autoregulatory mechanisms.

Advances in ultrathin, skin-like electronics lead to wearable devices for continuous, noninvasive blood flow monitoring., Continuous monitoring of variations in blood flow is vital in assessing the status of microvascular and macrovascular beds for a wide range of clinical and research scenarios. Although a variety of techniques exist, most require complete immobilization of the subject, thereby limiting their utility to hospital or clinical settings. Those that can be rendered in wearable formats suffer from limited accuracy, motion artifacts, and other shortcomings that follow from an inability to achieve intimate, noninvasive mechanical linkage of sensors with the surface of the skin. We introduce an ultrathin, soft, skin-conforming sensor technology that offers advanced capabilities in continuous and precise blood flow mapping. Systematic work establishes a set of experimental procedures and theoretical models for quantitative measurements and guidelines in design and operation. Experimental studies on human subjects, including validation with measurements performed using state-of-the-art clinical techniques, demonstrate sensitive and accurate assessment of both macrovascular and microvascular flow under a range of physiological conditions. Refined operational modes eliminate long-term drifts and reduce power consumption, thereby providing steps toward the use of this technology for continuous monitoring during daily activities.

Continuous infrared imaging revealed transient changes in forearm temperature during arterial occlusion, reperfusion, and recovery in a healthy subject group. Processing the imaging data with the k-means algorithm further revealed reactive vascular sites in the skin with rapid or delayed temperature amplification. The observed temporal and spatial diversity of blood-flow-derived forearm temperature allow consideration of thermal-imaging guided placement of skin sensors to achieve enhanced sensitivity in monitoring of skin hemodynamics.

International audience; Important progress occurred in pulse-stimulated thermography, in particular thanks to the TSR technique, a technique based on the decomposition of thermograms on a logarithmic polynomial basis and the use of the logarithmic derivatives to enhance the detection of defects in structures. Its fields of application begin to broaden to the characterization of transient internal heat sources in experimental mechanics and biomedicine. The TSR technique is presented, in particular the last developments leading to the production of a unique synthetic image. Two recent examples of applications in experimental mechanics and biomedicine, taken from literature, are described: in situ detection of damages in a composite material during mechanical tests and in vivo visualization of subcutaneous functional angioarchitecture in humans.

Heat liberation in the brain was utilized as a direct signature of functional activation. We hypothesize that both temporal and spatial uncoupling between local cerebral blood flow (lCBF) and metabolic temperature components can be explored through the imaging of brain thermal gradients evoked by functional stimulation. Surface cortical infrared (IR) images were obtained from 34 patients undergoing surgery for brain lesions under baseline conditions following peripheral nerve stimulation and, in some patients, during active behavioral tasks such as finger apposition and repetitive hand movements. An IR camera (0.02 °C sensitivity, 3–5 μm wavelength) was used to image local thermal gradients across the cerebral cortex by passively detecting IR emission. Neural activation elicited reproducible temperature changes (0.04–0.09 °C) within the primary somatosensory cortex during median nerve stimulation and in the sensorimotor cortex during repetitive hand movements and finger tapping. The initial temperature responses were detected as early as 100–200 ms, the peak IR response occurred 5–7 s after stimulus onset. Models of the relationship between evoked thermal gradients, lCBF and metabolic heat are proposed. Since the latencies of local metabolic and lCBF responses to stimulation vary by more than an order of magnitude, we are able to separate vascular-dependent and metabolic-dependent temperature components and thus create two discrete brain images, each reflecting distinct physiological mechanisms of functional activation. The resultant temperature profile reflects the balance between metabolism and lCBF, and therefore the degree of their functional uncoupling for the exposed and (possibly) for the intact normal human brain.

We hypothesized that it would be possible to detect the distribution of cortical activation by using a sensitive, rapid, high-resolution infrared imaging technique to monitor changes in local cerebral blood flow induced by changes in focal cortical metabolism. In a prospective study, we recorded in 21 patients the emission of infrared radiation from the exposed human cerebral cortex at baseline, during language and motor tasks, and during stimulation of the contralateral median nerve using an infrared camera (sensitivity 0.02 degrees C). The language and sensorimotor cortex was identified by standard mapping methods (cortical stimulation, median nerve somatosensory-evoked potential, functional magnetic resonance imaging), which were compared with infrared functional localization. The temperature gradients measured during surgery are dominated by changes in local cerebral blood flow associated with evoked functional activation. The distribution of the evoked temperature changes overlaps with, but extends beyond, functional regions identified by standard mapping techniques. The distribution observed via infrared mapping is consistent with distributed and complex functional representation of the cerebral cortex, rather than the traditional concept of discrete functional loci demonstrated by brief cortical stimulation during surgery and by noninvasive functional imaging techniques. By providing information on the spatial and temporal patterns of sensory-motor and language representation, infrared imaging may prove to be a useful approach to study brain function.

The present study was aimed to assess blood flow oscillatory patterns in rat pial microcirculation by laser speckle imaging. Laser speckle methods provides maps of cerebral blood flow with elevated temporal and spatial resolution. The study was carried out on male Wistar rats. An open cranial window was prepared on the parietal region. Pial arterioles were classified by Strahler method in five orders by fluorescence microscopy. The blood flow oscillatory patterns were determined by laser speckle imaging in single pial vessels. Power spectrum analysis was performed by wavelet methods under baseline conditions. Arterioles and venules were characterized by blood flow oscillations with frequency components in the ranges 0.005-0.0095 Hz, 0.0095-0.02 Hz, 0.02-0.15 Hz, 0.15-0.5 Hz. Arterioles showed oscillations with higher total power and higher spectral density in the range 0.02-0.15 when compared with venules. Laser speckle imaging allowed us to evaluate arteriolar and venular blood flow oscillations. ?????? 2014 IEEE.

Sickle cell disease is an inherited blood disorder characterized by chronic hemolytic anemia and episodic vaso-occlusive pain crises. Vaso-occlusion occurs when deoxygenated hemoglobin S polymerizes and erythrocytes sickle and adhere in the microvasculature, a process dependent on the concentration of hemoglobin S and the rate of deoxygenation, among other factors. We measured oxygen consumption in the thenar eminence during brachial artery occlusion in sickle cell patients and healthy individuals. Microvascular oxygen consumption was greater in sickle cell patients than in healthy individuals (median [interquartile range]; sickle cell: 0.91 [0.75-1.07] vs healthy: 0.75 [0.62-0.94] -ΔHbO2/min, P < .05) and was elevated further during acute pain crisis (crisis: 1.10 [0.78-1.30] vs recovered: 0.88 [0.76-1.03] -ΔHbO2/min, P < .05). Increased microvascular oxygen consumption during pain crisis could affect the local oxygen saturation of hemoglobin when oxygen delivery is limiting. Identifying the mechanisms of elevated oxygen consumption during pain crisis might lead to the development of new therapeutic interventions. This trial was registered at www.clinicaltrials.gov as #NCT01568710.

The disruption of the temperature circadian rhythm has been associated with cancer progression, while its amplification resulted in cancer inhibition in experimental tumor models. The current study investigated the relevance of skin surface temperature rhythms as biomarkers of the Circadian Timing System (CTS) in order to optimize chronotherapy timing in individual cancer patients. Baseline skin surface temperature at four sites and wrist accelerations were measured every minute for 4 days in 16 patients with metastatic gastro-intestinal cancer before chronotherapy administration. Temperature and rest-activity were recorded, respectively, with wireless skin surface temperature patches (Respironics, Phillips) and an actigraph (Ambulatory Monitoring). Both variables were further monitored in 10 of these patients during and after a 4-day course of a fixed chronotherapy protocol. Collected at baseline, during and after therapy longitudinal data sets were processed using Fast Fourier Transform Cosinor and Linear Discriminant Analyses methods. A circadian rhythm was statistically validated with a period of 24 h (p|0.7|; p

The hallmark manifestation of homozygous sickle cell disease (SCD) is acute pain. The underlying etiology is unknown. It is believed that hypoxia induces changes in erythrocyte shape and produces microvasculature occlusion by sickled red cells. Vascular occlusion is believed to represent the event inducing tissue hypoxia, reperfusion injury and pain. Studies have only recently characterized pain responses to experimental stimuli in rodents and humans with SCD using quantitative sensory testing (QST). SCD is characterized by thermal and mechanical sensitivity, and there is evidence of central sensitization (heightened sensitivity to pain) from QST, temporal summation and functional MRI studies. However, the effect of hypoxia upon nociception has not been studied despite the need for hypoxia to induce sickle hemoglobin polymerization. The ischemic tourniquet pain test is a QST used to investigate nociception. It was the first QST to demonstrate opioid analgesia. While it is assumed that the resulting pain is due to hypoxia, no studies have demonstrated the degree of hypoxia needed to produce pain. Methods: Thirty adults with SCD and 30 age and sex matched 30 controls underwent the ischemic pain test. A pneumatic cuff was inflated around the upper arm to induce forearm hypoxia, and the time from cuff inflation to first reported pain (pain threshold) and until pain became intolerable (pain tolerance) was measured. Pain was quantified on a 20 point scale. Time to pain threshold and tolerance were the primary end points. Testing was repeated after a washout period in a subset of the initial subjects: 18 SCD patients and 22 controls. Repeat testing proceeded with a 15 minute observation period followed by the ischemic pain test with the time and pain score recorded at threshold and tolerance. Subjects were monitored after cuff deflation during recovery with pain scores obtained at 30 seconds, then every minute for 5 minutes and finally every 3 minutes over a total recovery period of 20 minutes. Skeletal muscle tissue oxygenation at the thenar eminence was measured continuously with a near infrared spectroscopy (NIRS) monitor (OxiplexTS; ISS, Champaign, IL). Results: SCD subjects reached the pain threshold at a mean of 6.44 versus 11.57 minutes in controls (p=0.07). Time to pain tolerance was significantly different (SS 11.29 versus controls 19.59 minutes; p=0.004). Both groups reported identical pain scores. By stepwise linear regression, SCD (p=0.002) and gender (p=0.0008) were associated with time to pain tolerance (r2=0.29), while recent opioid use and pain crisis frequency were not. Because these results demonstrated that SCD adults reach tolerance more rapidly than controls, we sought to determine if this difference was due to tissue hypoxia. Pain scores with repeat testing were not significantly different at threshold or tolerance. However, pain score curves over testing and recovery were significantly different (p=0.003, Fig 1A). SCA had pain above baseline after 11 minutes of recovery (p=0.02) suggesting a persistent pain response compared to controls. Oxyhemoglobin and deoxyhemoglobin from NIRS were not different between groups at the primary endpoints (Fig 1B and 1C). StO2 (% tissue oxygenation, Fig 1D) at both threshold and tolerance was not different between SCD and controls, suggesting that the onset and maximal level of pain during this QST is defined an StO2 thresholds 40% common to sickle cell patients and controls. However, oxyhemoglobin, deoxyhemoglobin and StO2 had already recovered to baseline by the time the pain was evident (Fig 1B, 1C and 1D). Conclusions: Adults with SCD reach hypoxia induced, experimental pain earlier than NVs. Time to maximal pain is associated with SCD and sex. NIRS showed that pain tolerance occurred at a uniform tissue oxygenation threshold indicating hypoxia is the primary determinant for sensing acute pain. However, persistent SCD pain was independent of tissue oxygenation, and could be explained by the presence of peripheral or central sensitization. These data suggest that SCD pain treatment may require improved oxygen delivery to affected tissues and analgesics. Figure 1. Nociceptive and NIRS responses to ischemic pain testing in adults with sickle cell anemia. Figure 1. Nociceptive and NIRS responses to ischemic pain testing in adults with sickle cell anemia. Disclosures No relevant conflicts of interest to declare.

Precision thermometry of the skin can, together with other measurements, provide clinically relevant information about cardiovascular health, cognitive state, malignancy and many other important aspects of human physiology. Here, we introduce an ultrathin, compliant skin-like sensor/actuator technology that can pliably laminate onto the epidermis to provide continuous, accurate thermal characterizations that are unavailable with other methods. Examples include non-invasive spatial mapping of skin temperature with millikelvin precision, and simultaneous quantitative assessment of tissue thermal conductivity. Such devices can also be implemented in ways that reveal the time-dynamic influence of blood flow and perfusion on these properties. Experimental and theoretical studies establish the underlying principles of operation, and define engineering guidelines for device design. Evaluation of subtle variations in skin temperature associated with mental activity, physical stimulation and vasoconstriction/dilation along with accurate determination of skin hydration through measurements of thermal conductivity represent some important operational examples.

In this paper we present a long-term temperature monitoring system that uses miniature wearable sensor nodes that connect via Bluetooth to a cellular phone in a star topology. Monitoring of skin temperature over multiple days has been shown to be useful in clinical research related to metabolic and cardiovascular diseases, and it has a potential role in the study of circadian rhythms in patients with cancer. The system can provide immediate remote feedback through a phone's internet connectivity and eliminates the need for a dedicated base station. By utilizing Bluetooth in a burst-mode regime, sensor nodes can work autonomously for 135 hours at a sampling rate of 0.2 Hz. A validation study showed that the system can reliably record circadian temperature rhythms in an ambulatory environment, and can serve as a sensor platform for novel applications.

We demonstrate a new mid-infrared and near-infrared imaging approach which is ideally suited to microscopic applications. The method employs an indium antimonide (InSb) focal-plane array detector and a commercially available step-scan Fourier transform infrared spectrometer (FT-IR). With either a KBr or a CaF2beamsplitter, images from 1 to 5.5 μm (10,000-1818 cm−1) can be rapidly acquired with the use of all the available pixels on the detector. The spectral resolution for each image is easily varied by changing the number of acquired images during the interferometer scan. We apply this technique to noninvasively generate image contrast in sections of monkey brain tissue and to relate these data to specific lipid and protein fractions. In addition, we describe several computational methods to highlight the spatial distributions of components within a sample.

The clinical hallmark of sickle cell anemia is the vaso-occlusive pain crisis. Although the exact cause for severe vaso-occlusive painful events is unknown, sickle cell microvasculature occlusion is thought to be the proximate cause producing tissue hypoxia, reperfusion injury and acute pain. Endothelial dysfunction is a prominent characteristic of sickle cell anemia, and it is unclear to what extent this abnormal vascular response contributes to vaso-occlusion and pain. We sought to evaluate the effects of hypoxia on sickle cell pain by performing a forearm ischemic pain test as a potential in vivo model for vaso-occlusion. We hypothesized that sickle cell anemia patients would tolerate a shorter period of ischemia before reaching pain tolerance. We further hypothesized that sickle patients would show more hypoxia and increased vasodilation. Thirty adults with sickle cell anemia were recruited and matched by age and sex to 30 normal volunteers. We first performed a timed ischemic pain test with brachial artery occlusion until subjects first reported pain (pain threshold) and until maximum pain tolerated (pain tolerance). Sickle cell subjects first reported pain at 411 vs. 589 s for normal volunteers (mean, p=0.07). Occlusion time to pain tolerance was significantly shorter for sickle cell patients (637 vs. 918 s, mean, p=0.004). Despite this difference, both groups reported nearly identical pain scores at threshold and tolerance. Stepwise linear regression for all subjects against 8 variables likely to influence pain showed sickle status (p=0.002) and gender (p=0.0008) were independently associated with time to tolerance, supporting our initial hypothesis. Testing with continuous physiological monitoring was next repeated in sub-groups of 7 sickle cell and 9 normal subjects in an effort to understand the association between ischemia and pain progression. Before, during, and after brachial artery occlusion, oxygenated/deoxygenated hemoglobin concentration and tissue oxygen saturation were continuously monitored with near-infrared spectroscopy at the thenar eminence. We also recorded cutaneous blood flow with a Laser Speckle Contrast Imager (FLPI-2) in the volar aspect of forearm and continuous blood pressure and pulse in the contralateral arm. Monitoring was performed during steady state prior to occlusion (15 min), during occlusion until pain tolerance, and during recovery (20 min). At steady state, sickle cell subjects had higher median heart rate (68 vs. 62 bpm, p=0.05) and cutaneous blood flow (81.8 vs. 46.8 a.u., p Overall, sickle cell subjects have significantly lower steady state tissue oxygenation, but they are less tolerant of hypoxia and develop pain at higher oxygenated hemoglobin levels during ischemia. Despite higher oxygenated hemoglobin during ischemia, sickle cell subjects have a significantly higher absolute decline in blood flow during occlusion, suggesting an altered hypoxic response compared to controls. This might suggest a hypersensitive hypoxic pain response, possibly due to the presence of chronic pain, and altered oxygen sensing. The ischemic pain test is a potential in vivo model for early stage trials of drugs that alter either acute pain transmission or oxygen delivery to tissues. Disclosures No relevant conflicts of interest to declare.

Background: A lower core body temperature set point has been suggested to be a factor that could potentially predispose humans to develop obesity. Objective: We tested the hypothesis that obese individuals have lower core temperatures than those in normal-weight individuals. Design: In study 1, nonobese [body mass index (BMI; in kg/m2)

Assessment of pulsatile perfusion (PP) is limited to measurements of flow (V) and resistance (R). We investigated infrared (IR) imaging during PP as a means for precise organ assessment. IR was used to monitor 10 porcine kidneys during 18 hr of PP in an uncontrolled Donation after Cardiac Death model. An IR camera (Lockheed Martin) was focused on the anterior surfaces of the kidneys. The degree of temperature homogeneity was compared with standard measurements of V and R. IR thermal images correlated with V and R (R=0.92, P0.001). IR detected an increase in homogeneity during PP by comparing standard deviation differences before and after PP (P=0.002), which was not evident by standard measurements of V and R. Finally, IR assessment allowed for measurement of dynamic changes in perfusion.

To assess vascular responses of the human hand to inspiratory gasps and hand cooling, two imaging "remote sensing" instruments were utilized: 1) a high-resolution infrared (IR) imaging camera and 2) a full-field laser perfusion imager (FLPI). Data analysis was performed on the data sets collected simultaneously from both instruments. A non-localized drop of both FLPI and IR signals was observed at ~0.5-2.0 min after gasp onset. Spontaneous oscillations, much below the human cardiac and respiratory frequencies, were observed with both imagers. The dominant oscillations for both imaging modalities centered around 0.01Hz. Spectral frequencies, their power, and the duration of temperature oscillations (bursts) for different hand areas changed in time and were spatially heterogeneous. The highest spatial correlation between the two data sets was found between the mean IR derivative image and the mean original FLPI image for the baseline conditions. Heterogeneous images of the human hand were consistently detected non-invasively by both instruments. After cooling, a temperature elevation of ~0.5oC was seen as a spotted pattern mainly in the thenar and hypothenar areas. A generalized increase in perfusion over the same areas was observed in FLPI images. Both IR and FLPI imagers sensitively identify vasoconstrictor responses induced by inspiratory gasp and hand cooling maneuvers. The specificity to physiological changes and high imaging rate for both instruments, coupled with the current ease of use of optical cameras in clinical settings, make the described combination of two instruments an ideal imaging approach to studying the dynamics of thermal and perfusion heterogeneity in human skin.

Complex challenges of optical imaging in diagnostics and surgical treatment require accurate image registration/stabilization methods that remove only unwanted motions. An SIAROI algorithm is proposed for real-time subpixel registration sequences of intraoperatively acquired infrared (thermal) brain images. SIAROI algorithm is based upon automatic, localized Subpixel Image Autocorrelation and a user-selected Region of Interest (ROI). Human expertise about unwanted motions is added through a user-outlined ROI, using a low-accuracy free-hand paintbrush. SIAROI includes: (a) propagating the user-outlined ROI by selecting pixels in the second image of the sequence, using the same ROI; (b) producing SROI (sub-pixel ROI) by converting each pixel to k=NxN subpixels; (c) producing new SROI in the second image by shifting SROI within plus or minus 6k subpixels; (d) finding an optimal autocorrelation shift (x,y) within 12N that minimizes the Standard Deviation of Differences of Pixel Intensities (SDDPI) between corresponding ROI pixels in both images, (e) shifting the second image by (x,y), repeating (a)-(e) for successive images (t,t1). In experiments, a user quickly outlined non-deformable ROI (such as bone) in the first image of a sequence. Alignment of 100 brain images (25600x25600 pixel search, after every pixel was converted to 100 sub-pixels), took ~3 minutes, which is 200 times faster (with a 0.1=ROI/image ratio) than global auto-correlation. SIAROI improved frame alignment by a factor of two, relative to a Global Auto-correlation and Tie-points-based registration methods, as measured by reductions in the SDDPI.

The National Institute of Standards and Technology and the National Institutes of Health have started a collaborative study on the development of lighting that will provide enhanced, tissue-specific contrast with respect to its surroundings. In this paper we describe existing NIST technologies utilized for this project such as a computational model for color rendering and a new spectrally tunable lighting technology. We will also describe the calibration and validation procedure of a hyperspectral camera system. Finally, we show examples of imaged tissues under various lighting conditions.

Intraoperative optical imaging of exposed organs in visible, near-infrared, and infrared (IR) wavelengths in the body has the potential to be useful for real-time assessment of organ viability and image guidance during surgical intervention. However, the motion of the internal organs presents significant challenges for fast analysis of recorded 2D video sequences. The movement observed during surgery, due to respiration, cardiac motion, blood flow, and mechanical shift accompanying the surgical intervention, causes organ reflection in the image sequence, making optical measurements for further analysis challenging. Correcting alignment is difficult in that the motion is not uniform over the image. This paper describes a Canny edge-based method for segmentation of the specific organ or region under study, along with a moment-based registration method for the segmented region. Experimental results are provided for a set of intraoperative IR image sequences.

Object. Although clinical imaging defines the anatomical relationship between a brain tumor and the surrounding brain and neurological deficits indicate the neurophysiological consequences of the tumor, the effect of a brain tumor on vascular physiology is less clear. Methods. An infrared camera was used to measure the temperature of the cortical surface before, during, and after removal of a mass in 34 patients (primary brain tumor in 21 patients, brain metastases in 10 and falx meningioma, cavernous angioma, and radiation necrosis—astrocytosis in one patient each). To establish the magnitude of the effect on blood flow induced by the tumor, the images were compared with those from a group of six patients who underwent temporal lobectomy for epilepsy. In four cases a cerebral artery was temporarily occluded during the course of the surgery and infrared emissions from the cortex before and after occlusion were compared to establish the relationship of local temperature to regional blood flow. Discrete temperature gradients were associated with surgically verified lesions in all cases. Depending on the type of tumor, the cortex overlying the tumor was either colder or warmer than the surrounding cortex. Spatial reorganization of thermal gradients was observed after tumor resection. Temperature gradients of the cortex in patients with tumors exceeded those measured in the cortex of patients who underwent epilepsy surgery. Conclusions. Brain tumors induce changes in cerebral blood flow (CBF) in the cortex, which can be made visible by performing infrared imaging during cranial surgery. A reduction in CBF beyond the tumor margin improves after removal of the lesion.

Allograft ischemia induces delayed graft function and is correlated with increasing rates of rejection. There is not currently a way to objectively measure the effects of ischemia in real-time, nor to relate therapies combating reperfusion injury with their intended effects. An infrared (IR) method utilizing a focal plane array detector camera was developed for real-time intraoperative IR imaging of renal allografts, and evaluated in a pilot trial to quantify perfusion in recipients of live (n = 8) and cadaveric donor (n = 5) allografts. Digital images were taken for 3–8 min postreperfusion. Image data were compared to ischemic time and allograft function to assess potential clinical relevance. Cold ischemic time ranged from 0.5 to 29 h and was bimodally distributed between living and cadaveric donors. Renal rewarming time (RT) as determined by IR imaging correlated with cold ischemic time (p

Object. Application of sensitive infrared imaging is ideally suited to observe blood vessels and blood flow in exposed organs, including the brain. Temporary vascular occlusion is an important part of neurosurgery, but the capacity to monitor the effects of these occlusions in real time is limited. In surgical procedures that require vascular manipulation, such as those involving aneurysms, arteriovenous malformations (AVMs), or tumors, the ability to visualize blood flow in vessels and their distribution beds would be beneficial. The authors recount their experience in the use of a sensitive (0.02°C), high-resolution (up to 50 µm/pixel) infrared camera with a rapid shutter speed (up to 2 msec/frame) for localizing cortical function intraoperatively. They observed high-resolution images of cerebral arteries and veins. The authors hypothesized that infrared imaging of cerebral arteries, performed using a sensitive, high-resolution camera during surgery, would permit changes in arterial flow to be be seen immediately, thus providing real-time assessment of brain perfusion in the involved vascular territory. Methods. Cynomolgus monkeys underwent extensive craniectomies, exposing the frontal, parietal, and temporal lobes. Temporary occlusions of the internal carotid artery and middle cerebral artery branches (30 events) were performed serially and were visualized with the aid of an infrared camera. Arteries and veins of the monkey brain were clearly visualized due to cooling of the exposed brain, which contrasted with blood within the vessels that remained at core temperature. Blood flow changes in vessels were seen immediately (< 1 second) in real time during occlusion and reopening of the vessels, regardless of the duration of the occlusion. Areas of decreased cortical blood flow rapidly cooled (−0.3 to 1.3°C) and reheated in response to reperfusion. Rewarming occurred faster in arteries than in the cortex (for a 20-minute occlusion, the change in temperature per second was 2 × 10−2°C in the artery and 7 × 10−3°C in the brain). Collateral flow could be evaluated by intraoperative observations and data processing. Conclusions. Use of high-resolution, digital infrared imaging permits real-time visualization of arterial flow. It has the potential to provide the surgeon with a means to assess collateral flow during temporary vessel occlusion and to visualize directly the flow in parent arteries or persistent filling of an aneurysm after clipping. During surgery for AVMs, the technique may provide a new way to assess arterial inflow, venous outflow, results of embolization, collateral flow, steal, and normal perfusion pressure breakthrough.

Thermal representations on the surface of a human forearm of underlying perforator vessels have previously been mapped via recovery-enhanced infrared imaging, which is performed as skin blood flow recovers to baseline levels following cooling of the forearm. We noted that the same vessels could also be observed during reactive hyperaemia tests after complete 5-min occlusion of the forearm by an inflatable cuff. However, not all subjects showed vessels with acceptable contrast. Therefore, we applied a thermographic signal reconstruction algorithm to reactive hyperaemia testing, which substantially enhanced signal-to-noise ratios between perforator vessels and their surroundings, thereby enabling their mapping with higher accuracy and a shorter occlusion period.

Background While a number of the consequences of mast cell degranulation within tissues have been documented including tissue-specific changes such as bronchospasm and the subsequent cellular infiltrate, there is little known about the immediate effects of mast cell degranulation on the associated vasculature, critical to understanding the evolution of mast cell dependent inflammation. Objective To characterize the microcirculatory events that follow mast cell degranulation. Methodology/principal findings Perturbations in dermal blood flow, temperature and skin color were analyzed using laser-speckle contrast imaging, infrared and polarized-light colorimetry following cold-hand immersion (CHI) challenge in patients with cold-induced urticaria compared to the response in healthy controls. Evidence for mast cell degranulation was established by documentation of serum histamine levels and the localized release of tryptase in post-challenge urticarial biopsies. Laser-speckle contrast imaging quantified the attenuated response to cold challenge in patients on cetirizine. We found that the histamine-associated vascular response accompanying mast cell degranulation is rapid and extensive. At the tissue level, it is characterized by a uniform pattern of increased blood flow, thermal warming, vasodilation, and recruitment of collateral circulation. These vascular responses are modified by the administration of an antihistamine. Conclusions/significance Monitoring the hemodynamic responses within tissues that are associated with mast cell degranulation provides additional insight into the evolution of the acute inflammatory response and offers a unique approach to assess the effectiveness of treatment intervention.

Abstract 1080 Background: The pathophysiology of sickle cell disease (SCD) involves vascular complications such as stroke and pulmonary hypertension. Elevated pulmonary artery pressure estimated by cardiac ultrasound or measured by invasive right heart catheterization is associated with early mortality among patients with SCD. Peripheral vascular dysfunction has also been observed in SCD and may be easier to assess than pulmonary vascular resistance and therefore more suitable for epidemiologic or interventional studies of SCD. Three previous studies found no difference in the maximal blood flow (or percentage increase) stimulated by occlusion and reperfusion of the brachial artery, a conduit vessel, when measured by Doppler ultrasound at periodic intervals. However, a study using continuous laser Doppler measurements of cutaneous microvascular blood flow found that patients with SCD had prolonged time to maximal blood flow and prolonged time to return to baseline compared to healthy controls. In our study, we used two-dimensional laser speckle contrast imaging (LSCI) to assess the cutaneous microvascular blood flow response in adults with SCD after five minutes of brachial artery occlusion. Methods: Nine subjects with sickle cell disease were enrolled and compared against nine healthy African-American control subjects matched for age, sex, ethnicity, and body mass index. Cutaneous blood flow was directly measured using LCSI at baseline, during and after a standard brachial artery occlusion-reperfusion maneuver (inflation of an occlusive pneumatic cuff for five minutes). This stimulates a transient increase in blood flow to levels above baseline during the reperfusion phase. Blood flow data were averaged over a defined region of interest on the medial aspect of the forearm. Microvascular blood flow responsiveness was calculated as the time to maximum (time elapsed from 50% of maximum to maximum blood flow) and the time to return to baseline (time elapsed from maximum to 50% of maximum blood flow). We performed measurements on each individual on two separate days, and compared the microvascular blood flow responses between the groups using two-way ANOVA with repeated measures. Results: We enrolled nine patients with sickle cell disease (age 35 ± 8.8, BMI 23 ± 3.9, 3 men/6 women) and nine healthy controls (age 35 ± 10.7 years, BMI 25 ± 3.2, 3 men/6 women). Baseline microvascular blood flow measured in arbitrary units (AU) was greater in patients with sickle cell disease compared to healthy controls (53.1 ± 9.2 AU vs 37.2 ± 4.4 AU, p < 0.0001) but maximal microvascular blood flow was similar (121.3 ± 29.3 AU vs 124.7 ± 26.6 AU, p = 0.58). Analysis of the time from half-maximum to maximum blood flow revealed that patients with sickle cell disease take longer to reach maximum blood flow (19.1 ± 11.6 s vs 11.8 ± 1.0 s, p = 0.03) and longer to decrease from maximum to half-maximum blood flow during the recovery period (43.5 ± 13.0 s vs 28.6 ± 10.4 s, p = 0.002). Conclusion: Compared to healthy individuals, patients with SCD have greater baseline microvascular blood flow but similar maximal blood flow during reperfusion. However, patients with SCD differ significantly from healthy control subjects in the time required to reach maximal blood flow and the time required to return to baseline, both of which are prolonged in patients with SCD. This may reflect delayed or impaired endothelial responses to shear stress and/or greater viscosity of blood. Time to maximal blood flow might represent a useful physiological biomarker as a proxy for clinical severity of sickle cell anemia, and a potential surrogate marker in early phase clinical trials. This technique merits additional characterization and validation. Disclosures: No relevant conflicts of interest to declare.

Abstract 1009 Background: Leg ulcerations are a serious and debilitating complication of sickle cell disease (SCD) and other hemolytic anemias, whose etiology is not completely understood. Wound healing involves an ordered progression of vasoconstriction (hemostatic phase), followed by vasodilatation (inflammatory and proliferative phase) and epithelialization. Little is known about the changes in the microcirculation of chronic wounds in patients with sickle cell disease. Endothelial function affects microcirculation and regulates blood flow. Endothelial dysfunction is a recognized pathway to end organ dysfunction in SCD. Laser speckle contrast imaging (LSCI) and Infrared Thermography (IR) are non-invasive technique used to assess dynamic changes in blood flow in response to vaso-reactive and other insults in both health and disease, while peripheral arterial tonometry is utilized for early detection of cardiovascular disease. The aim of this study was to evaluate skin's microvasculature response and endothelial function in adults with HbSS and chronic leg ulceration. Methods: Twelve adults with HbSS and chronic leg ulcers (> 1 month duration) were evaluated at steady state with a detailed medical and ulcer history, physical examination, and clinical labs. Clinical and laboratory data were compared to 269 adults with Hb SS, without active leg ulcers, enrolled in a Pulmonary Hypertension screening protocol (Clin trial no. NCT00011648). Blood flow (LCSI) and IR measurements of the ulcer bed, the peri-ulcer area and of a distant, unaffected area (baseline) were obtained and compared. Peripheral arterial tonometry (PAT) was obtained to calculate the ratio of post-occlusion dilatation to baseline reactive hyperemia index: RHI. An RHI>1.67 is indication of normal endothelial function. Results: Mean age for the 12 subjects with chronic active leg ulcer: 31 years ± 11yrs, 5 male, 5 had a history of trauma. When compared to SS patients without active leg ulcers: mean MAP 82 vs 103 mmHg (p Conclusions: With this study we confirm that patients with HbSS and leg ulcers represent a subgroup with severe disease, as demonstrated by the high use of HU and/or transfusion, and number of complications at a relatively young age. We report high blood flow in the ulcer bed and immediate surrounding regions, compared to unaffected areas, never prior demonstrated. It is probable that the increase in blood flow observed in the wound and peri-wound regions reflect s a high vasodilated state, resulting from severe anemia, relative hypotension, and chronic inflammation. Laboratory and clinical data supports the presence of a persistent inflammatory and hemolytic state, as well as low arginine bioavailabitlity. Laser Speckle Contrast Imaging, Infrared Thermography and Peripheral Arterial Tonometry are effective, non invasive novel techniques useful for studying microvascular and endothelial function in patients with SCD and leg ulcers. Disclosures: No relevant conflicts of interest to declare.

Abstract 86 Background: Leg ulcers are a common and debilitating complication of sickle cell anemia and other hemolytic disorders. Despite many advances in the care of SCD, there is still a clinical need for a well-tolerated, safe and effective therapy. Sodium nitrite may function as a reservoir for local delivery of nitric oxide, whose vasodilating, angiogenic, and antimicrobicidal activities make it an attractive novel therapy for chronic wounds. We describe the safety data and preliminary efficacy data of topical sodium nitrite in adults with sickle cell anemia. Design and Methods: This is a phase 1 dose-escalation, single institution, study of topical sodium nitrite (ClinicalTrials.gov NCT01316796). Eligibility criteria: > 18 years old, have a leg ulcer of > 4 weeks duration, between 2.5 and 100 cm2 in size, and not acutely infected. Patients were screened with medical history, physical exam and laboratory tests. Sodium nitrite cream was applied twice a week for 4 weeks on one leg ulcer (“study ulcer”) in four cohorts of three subjects each with escalating concentrations in each cohort of 0.5., 1, 1.5 and 2%. Safety, tolerability and pharmacokinetic data of plasma nitrite and nitrate were obtained during the first 48 hours of drug exposure and then weekly for four weeks. Leg ulcer healing was a secondary endpoint and was assessed by calculating the change in surface area from visible light photographs of the study leg ulcer obtained at week 1, 3, and end of study, using a digital camera at a distance of 0.25 – 0.5 m from the ulcer. Borders were traced using ENVI software and ulcer size was calculated by converting the pixel area of the ulcer to cm2 using a calibrated square for reference. Pain at the ulcer site was assessed both with a Brief Pain Inventory and Visual Analog Pain Scale at predetermined time points. Results: Sixteen subjects were screened and 12 enrolled. There were no serious adverse events associated with the drug. Possibly related, grade 1; adverse events occurred in 10 of the subjects, more frequently in the cohorts with the highest concentration, which included non-clinically significant decrease in diastolic blood pressure in two subjects of cohort 4. Adverse events resolved without clinical intervention. Methemoglobin levels did not exceed pre-established safety thresholds (max of 4.1 % in one subject in cohort 3). Tolerability was excellent, with short-lived stinging at the site of application reported by two subjects. Pharmacokinetics of plasma nitrite and nitrate indicated minimal systemic absorption of topically applied sodium nitrite (median plasma nitrite AUC: 0.311 (0.169–0.659) umol*h/L/umol nitrite dose), with high interpatient variability. There was no evidence of plasma nitrite and nitrate and methemoglobin accumulation during the 4- week study trial. All but one subject experienced a decrease in leg ulcer surface area, fig 1, (pretreatment 4.65 vs. post-treatment 2.78 cm2, p Conclusions: On the basis of these safety, pharmacokinetic and tolerability data, and promising efficacy results, topical sodium nitrite warrants further clinical evaluation in patients with sickle cell disease or other hemolytic disorders and leg ulcers. Disclosures: No relevant conflicts of interest to declare.

The electrical potential produced by a dipole in the temporal or frontal lobe was calculated for a realistically shaped scalp, skull, and brain. This potential distribution was then used with a 3-sphere model to predict the position, orientation, and strength of the dipole source. The original and predicted dipole positions differed by an average of 1.97 cm, with a difference of more than 4 cm in some cases. Control calculations demonstrated that this difference was not caused by numerical artifacts in the computation, but instead was due to a true difference between the 3-sphere and realistically shaped head models.