Multi-Aspect Optoacoustic Imaging of Breast Tumors under Chemotherapy with Exogenous and Endogenous Contrasts: Focus on Apoptosis and Hypoxia

Breast cancer is a complex tumor type involving many biological processes. Most chemotherapeutic agents exert their antitumoral effects by rapid induction of apoptosis. Another main feature of breast cancer is hypoxia, which may drive malignant progression and confer resistance to various forms of therapy. Thus, multi-aspect imaging of both tumor apoptosis and oxygenation in vivo would be of enormous value for the effective evaluation of therapy response. Herein, we demonstrate the capability of a hybrid imaging modality known as multispectral optoacoustic tomography (MSOT) to provide high-resolution, simultaneous imaging of tumor apoptosis and oxygenation, based on both the exogenous contrast of an apoptosis-targeting dye and the endogenous contrast of hemoglobin. MSOT imaging was applied on mice bearing orthotopic 4T1 breast tumors before and following treatment with doxorubicin. Apoptosis was monitored over time by imaging the distribution of xPLORE-APOFL750©, a highly sensitive poly-caspase binding apoptotic probe, within the tumors. Oxygenation was monitored by tracking the distribution of oxy- and deoxygenated hemoglobin within the same tumor areas. Doxorubicin treatment induced an increase in apoptosis-depending optoacoustic signal of xPLORE-APOFL750© at 24 h after treatment. Furthermore, our results showed spatial correspondence between xPLORE-APO750© and deoxygenated hemoglobin. In vivo apoptotic status of the tumor tissue was independently verified by ex vivo fluorescence analysis. Overall, our results provide a rationale for the use of MSOT as an effective tool for simultaneously investigating various aspects of tumor pathophysiology and potential effects of therapeutic regimes based on both endogenous and exogenous molecular contrasts.

Prefrontal Cortex Activation During Motor Sequence Learning Under Interleaved and Repetitive Practice: A Two-Channel Near-Infrared Spectroscopy Study

Training under high interference conditions through interleaved practice (IP) results in performance suppression during training but enhances long-term performance relative to repetitive practice (RP) involving low interference. Previous neuroimaging work addressing this contextual interference effect of motor learning has relied heavily on the blood-oxygen-level-dependent (BOLD) response using functional magnetic resonance imaging (fMRI) methodology resulting in mixed reports of prefrontal cortex (PFC) recruitment under IP and RP conditions. We sought to clarify these equivocal findings by imaging bilateral PFC recruitment using functional near-infrared spectroscopy (fNIRS) while discrete key pressing sequences were trained under IP and RP schedules and subsequently tested following a 24-h delay. An advantage of fNIRS over the fMRI BOLD response is that the former measures oxygenated and deoxygenated hemoglobin changes independently allowing for assessment of cortical hemodynamics even when there is neurovascular decoupling. Despite slower sequence performance durations under IP, bilateral PFC oxygenated and deoxygenated hemoglobin values did not differ between practice conditions. During test, however, slower performance from those previously trained under RP coincided with hemispheric asymmetry in PFC recruitment. Specifically, following RP, test deoxygenated hemoglobin values were significantly lower in the right PFC. The present findings contrast with previous behavioral demonstrations of increased cognitive demand under IP to illustrate a more complex involvement of the PFC in the contextual interference effect. IP and RP incur similar levels of bilateral PFC recruitment, but the processes underlying the recruitment are dissimilar. PFC recruitment during IP supports action reconstruction and memory elaboration while RP relies on PFC recruitment to maintain task variation information in working memory from trial to trial. While PFC recruitment under RP serves to enhance immediate performance, it does not support long-term performance.

In Vivo Transcutaneous Monitoring of Hemoglobin Derivatives Using a Red-Green-Blue Camera-Based Spectral Imaging Technique

Cyanosis is a pathological condition that is characterized by a bluish discoloration of the skin or mucous membranes. It may result from a number of medical conditions, including disorders of the respiratory system and central nervous system, cardiovascular diseases, peripheral vascular diseases, deep vein thrombosis, and regional ischemia. Cyanosis can also be elicited from methemoglobin. Therefore, a simple, rapid, and simultaneous monitoring of changes in oxygenated hemoglobin and deoxygenated hemoglobin is useful for protective strategies against organ ischemic injury. We previously developed a red-green-blue camera-based spectral imaging method for the measurements of melanin concentration, oxygenated hemoglobin concentration (CHbO), deoxygenated hemoglobin concentration (CHbR), total hemoglobin concentration (CHbT) and tissue oxygen saturation (StO2) in skin tissues. We leveraged this approach in this study and extended it to the simultaneous quantifications of methemoglobin concentration (CmetHb), CHbO, CHbR, and StO2. The aim of the study was to confirm the feasibility of the method to monitor CmetHb, CHbO, CHbR, CHbT, and StO2. We performed in vivo experiments using rat dorsal skin during methemoglobinemia induced by the administration of sodium nitrite (NaNO2) and changing the fraction of inspired oxygen (FiO2), including normoxia, hypoxia, and anoxia. Spectral diffuse reflectance images were estimated from an RGB image by the Wiener estimation method. Multiple regression analysis based on Monte Carlo simulations of light transport was used to estimate CHbO, CHbR, CmetHb, CHbT, and StO2. CmetHb rapidly increased with a half-maximum time of less than 30 min and reached maximal values nearly 60 min after the administration of NaNO2, whereas StO2 dramatically dropped after the administration of NaNO2, indicating the temporary production of methemoglobin and severe hypoxemia during methemoglobinemia. Time courses of CHbT and StO2, while changing the FiO2, coincided with well-known physiological responses to hyperoxia, normoxia, and hypoxia. The results indicated the potential of this method to evaluate changes in skin hemodynamics due to loss of tissue viability and vitality.

Functional Photoacoustic and Ultrasonic Assessment of Osteoporosis: A Clinical Feasibility Study

Objective and Impact Statement. To study the feasibility of combined functional photoacoustic (PA) and quantitative ultrasound (US) for diagnosis of osteoporosis in vivo based on the detection of chemical and microarchitecture (BMA) information in calcaneus bone. Introduction. Clinically available X-ray or US technologies for the diagnosis of osteoporosis do not report important parameters such as chemical information and BMA. With unique advantages, including good sensitivity to molecular and metabolic properties, PA bone assessment techniques hold a great potential for clinical translation. Methods. By performing multiwavelength PA measurements, the chemical information in the human calcaneus bone, including mineral, lipid, oxygenated-hemoglobin, and deoxygenated-hemoglobin, were assessed. In parallel, by performing PA spectrum analysis, the BMA as an important bone physical property was quantified. An unpaired t-test and a two-way ANOVA test were conducted to compare the outcomes from the two subject groups. Results. Multiwavelength PA measurement is capable of assessing the relative contents of several chemical components in the trabecular bone in vivo, including both minerals and organic materials such as oxygenated-hemoglobin, deoxygenated-hemoglobin, and lipid, which are relevant to metabolic activities and bone health. In addition, PA measurements of BMA show good correlations (R2 up to 0.65) with DEXA. Both the chemical and microarchitectural measurements from PA techniques can differentiate the two subject groups. Conclusion. The results from this initial clinical study suggest that PA techniques, by providing additional chemical and microarchitecture information relevant to bone health, may lead to accurate and early diagnosis, as well as sensitive monitoring of the treatment of osteoporosis.

The effect of the fraction of inspired oxygen on the NIRS-derived deoxygenated hemoglobin “breakpoint” during ramp-incremental test

During ramp-incremental (RI) exercise to exhaustion, the near-infrared spectroscopy-derived deoxygenated hemoglobin ([HHb]) signal in the vastus lateralis muscle shows a linear increase up to a point at which a plateau-like response is manifested ([HHb]bp). This study investigated if 1) the [HHb]bp is affected by different fractions of inspired O2 ([Formula: see text]) [hypoxia (16%; HYPO); normoxia (21%; NORM); hyperoxia (30%; HYPER)]; and 2) an abrupt change to hyperoxic-inspired gas just before the occurrence of the [HHb]bp (HYPERSWITCH) would affect the [HHb] plateau-like response. Ten physically active male participants reported to the laboratory on four separate occasions to perform an RI test to exhaustion in NORM, HYPO, and HYPER and an RI test to exhaustion with an abrupt increase in [Formula: see text] (30%; HYPERSWITCH) 15 W before the power output (PO) associated with [HHb]bp in normoxia. PO, [HHb], tissue O2 ([Formula: see text]), and pulse O2 saturation ([Formula: see text]) were recorded continuously. Peak PO was significantly lower in HYPO (290 ± 21 W) and higher in HYPER (321 ± 22 W) and HYPERSWITCH (320 ± 19 W) compared with NORM (311 ± 18 W). The PO associated with [HHb]bp was not different between NORM and HYPER (246 ± 23 vs. 247 ± 24 W), but it was lower in HYPO (198 ± 31 W) than NORM and HYPER. The PO associated with the [HHb]bp in HYPERSWITCH (240 ± 23) was not different compared with NORM. HYPER and HYPERSWITCH resulted in greater [Formula: see text] and [Formula: see text] compared with NORM. These results suggest that the [HHb]bp response is not dependent of O2 driving pressure and that other physiological mechanisms might determine its occurrence.

Sequential Pulse Compression’s Effect on Blood Flow in the Lower-Extremity

Context: Several interventions are available to reduce the intensity and duration of the unwanted effects (eg, muscle soreness) associated with physical activity, such as massage, compression garments, and sequential pulse compression (SPC). Such interventions aim to increase blood flow to alleviate symptoms. However, there is a lack of evidence to support the use of SPC to alter total hemoglobin concentration (THb) in active individuals. Objective: To examine the acute effects of a single session of SPC on hemoglobin concentration compared with a control condition. Design: Single cohort, crossover design. Participants: Thirty-four physically active and healthy participants (females = 12 and males = 22) completed the study. Interventions: The authors randomly assigned participants to first receive the experimental (SPC) or control condition. Measures were recorded precondition and postcondition. Participants returned to the laboratory to complete the second condition ≥24 hours after the first condition. Main Outcome Measures: Relative changes in THb, deoxygenated hemoglobin, and oxygenated hemoglobin measures were recorded using near-infrared spectroscopy placed on the muscle belly of the medial gastrocnemius of the dominant limb. Results: SPC significantly increased THb (P < .001, d = 0.505) and oxygenated hemoglobin (P < .001, d = 0.745) change scores compared with the control condition. No statistical difference in deoxygenated hemoglobin change scores was found between the SPC and control conditions, but a medium effect size suggests potential biological significance (P = .06, d = 0.339). Conclusions: Overall, SPC increases THb to the lower-extremity and may be a viable option in the management of muscle soreness related to physical activity.

Association Between Deoxygenated Hemoglobin Breaking Point, Anaerobic Threshold, and Rowing Performance

Purpose: To compare the intensity and physiological responses of deoxygenated hemoglobin breaking point ([HHb]BP) and anaerobic threshold (AnT) during an incremental test and to verify their association with 2000-m rowing-ergometer performance in well-trained rowers. Methods: A total of 13 male rowers (mean [SD] age = 24 [11] y and = 63.7 [6.1] mL·kg−1·min−1) performed a step incremental test. Gas exchange, vastus lateralis [HHb], and blood lactate concentration were measured. Power output, , and heart rate of [HHb]BP and AnT were determined and compared with each other. A 2000-m test was performed in another visit. Results: No differences were found between [HHb]BP and AnT in the power output (236 [31] vs 234 [31] W; Δ = 0.7%), 95% confidence interval [CI] 6.7%), (4.2 [0.5] vs 4.3 [0.4] L·min−1; Δ = −0.8%, 95% CI 4.0%), or heart rate (180 [16] vs 182 [12] beats·min−1; Δ = −1.6%, 95% CI 2.1%); however, there was high typical error of estimate (TEE) and wide 95% limits of agreement (LoA) for power output (TEE 10.7%, LoA 54.1–50.6 W), (TEE 5.9%, LoA −0.57 to 0.63 L·min−1), and heart rate (TEE 2.4%, LoA −9.6 to 14.7 beats·min−1). Significant correlations were observed between [HHb]BP (r = .70) and AnT (r = .89) with 2000-m mean power. Conclusions: These results demonstrate a breaking point in [HHb] of the vastus lateralis muscle during the incremental test that is capable of distinguishing rowers with different performance levels. However, the high random error would compromise the use of [HHb]BP for training and testing in rowing.

Hyperspectral based discrimination of thyroid and parathyroid during surgery

Unintended injuring of anatomical structures during endocrine neck operations can have severe consequences for patient. Especially the nerves and the parathyroid gland can be hard to identify visually. Therefore, intraoperative methods are needed to support the surgeon in this task. Hyperspectral imaging (HSI) is a new approach in the medical area which combines a camera with a spectrometer. It showed promising results for the discrimination of tissue. In this work, HSI-data of seven patients were acquired during thyroid and parathyroid operations. The mean absorbance spectra of both glands showed differences in the range between 600 and 700 nm and at 760 and 960 nm. This means that thyroid and parathyroid have different oxygenation states and different contents of deoxygenated hemoglobin and water. From these observations, it is possible to define spectral signatures to characterize both glands. We showed on one patient how spectral signatures can be used in classification algorithms to automatically identify the thyroid and parathyroid from other structures.