Optical absorption spectra and corresponding in vivo photoacoustic visualization of exposed peripheral nerves.

Significance: Multispectral photoacoustic imaging has the potential to identify lipid-rich, myelinated nerve tissue in an interventional or surgical setting (e.g., to guide intraoperative decisions when exposing a nerve during reconstructive surgery by limiting operations to nerves needing repair, with no impact to healthy or regenerating nerves). Lipids have two optical absorption peaks within the NIR-II and NIR-III windows (i.e., 1000 to 1350 nm and 1550 to 1870 nm wavelength ranges, respectively) which can be exploited to obtain photoacoustic images. However, nerve visualization within the NIR-III window is more desirable due to higher lipid absorption peaks and a corresponding valley in the optical absorption of water. Aim: We present the first known optical absorption characterizations, photoacoustic spectral demonstrations, and histological validations to support in vivo photoacoustic nerve imaging in the NIR-III window. Approach: Four in vivo swine peripheral nerves were excised, and the optical absorption spectra of these fresh ex vivo nerves were characterized at wavelengths spanning 800 to 1880 nm, to provide the first known nerve optical absorbance spectra and to enable photoacoustic amplitude spectra characterization with the most optimal wavelength range. Prior to excision, the latter two of the four nerves were surrounded by aqueous, lipid-free, agarose blocks (i.e., 3% w/v agarose) to enhance acoustic coupling during in vivo multispectral photoacoustic imaging using the optimal NIR-III wavelengths (i.e., 1630 to 1850 nm) identified in the ex vivo studies. Results: There was a verified characteristic lipid absorption peak at 1725 nm for each ex vivo nerve. Results additionally suggest that the 1630 to 1850 nm wavelength range can successfully visualize and differentiate lipid-rich nerves from surrounding water-containing and lipid-deficient tissues and materials. Conclusions: Photoacoustic imaging using the optimal wavelengths identified and demonstrated for nerves holds promise for detection of myelination in exposed and isolated nerve tissue during a nerve repair surgery, with possible future implications for other surgeries and other optics-based technologies.

Comparison of compressional and elastic wave simulations for patient-specific planning prior to transcranial photoacoustic-guided neurosurgery.

SIGNIFICANCE: Simulations have the potential to be a powerful tool when planning the placement of photoacoustic imaging system components for surgical guidance. While elastic simulations (which include both compressional and shear waves) are expected to more accurately represent the physical transcranial acoustic wave propagation process, these simulations are more time-consuming and memory-intensive than the compressional-wave-only simulations that our group previously used to identify optimal acoustic windows for transcranial photoacoustic imaging. AIM: We present qualitative and quantitative comparisons of compressional and elastic wave simulations to determine which option is more suitable for preoperative surgical planning. APPROACH: Compressional and elastic photoacoustic k-Wave simulations were performed based on a computed tomography volume of a human cadaver head. Photoacoustic sources were placed in the locations of the internal carotid arteries and likely positions of neurosurgical instrument tips. Transducers received signals from three previously identified optimal acoustic windows (i.e., the ocular, nasal, and temporal regions). Target detectability, image-based target size estimates, and target-to-instrument distances were measured using the generalized contrast-to-noise ratio (gCNR), resolution, and relative source distances, respectively, for each simulation method. RESULTS: The gCNR was equivalent between compressional and elastic simulations. The areas of the -6 dB contours of point spread functions utilized to measure resolution differed by 0.33 to 3.35 mm2. Target-to-instrument distance measurements were within 1.24 mm of the true distances. CONCLUSIONS: These results indicate that it is likely sufficient to utilize the less time-consuming, less memory-intensive compressional wave simulations for presurgical planning.

Application of the generalized contrast-to-noise ratio to assess photoacoustic image quality.

The generalized contrast-to-noise ratio (gCNR) is a relatively new image quality metric designed to assess the probability of lesion detectability in ultrasound images. Although gCNR was initially demonstrated with ultrasound images, the metric is theoretically applicable to multiple types of medical images. In this paper, the applicability of gCNR to photoacoustic images is investigated. The gCNR was computed for both simulated and experimental photoacoustic images generated by amplitude-based (i.e., delay-and-sum) and coherence-based (i.e., short-lag spatial coherence) beamformers. These gCNR measurements were compared to three more traditional image quality metrics (i.e., contrast, contrast-to-noise ratio, and signal-to-noise ratio) applied to the same datasets. An increase in qualitative target visibility generally corresponded with increased gCNR. In addition, gCNR magnitude was more directly related to the separability of photoacoustic signals from their background, which degraded with the presence of limited bandwidth artifacts and increased levels of channel noise. At high gCNR values (i.e., 0.95-1), contrast, contrast-to-noise ratio, and signal-to-noise ratio varied by up to 23.7-56.2 dB, 2.0-3.4, and 26.5-7.6×1020, respectively, for simulated, experimental phantom, and in vivo data. Therefore, these traditional metrics can experience large variations when a target is fully detectable, and additional increases in these values would have no impact on photoacoustic target detectability. In addition, gCNR is robust to changes in traditional metrics introduced by applying a minimum threshold to image amplitudes. In tandem with other photoacoustic image quality metrics and with a defined range of 0 to 1, gCNR has promising potential to provide additional insight, particularly when designing new beamformers and image formation techniques and when reporting quantitative performance without an opportunity to qualitatively assess corresponding images (e.g., in text-only abstracts).

Photoacoustic Spatial Coherence Theory and Applications to Coherence-Based Image Contrast and Resolution.

The photoacoustic effect relies on optical transmission, which causes thermal expansion and generates acoustic signals. Coherence-based photoacoustic signal processing is often preferred over more traditional signal processing methods due to improved signal-to-noise ratios, imaging depth, and resolution in applications such as cell tracking, blood flow estimation, and imaging. However, these applications lack a theoretical spatial coherence model to support their implementation. In this article, the photoacoustic spatial coherence theory is derived to generate theoretical spatial coherence functions. These theoretical spatial coherence functions are compared with k-Wave simulated data and experimental data from point and circular targets (0.1-12 mm in diameter) with generally good agreement, particularly in the shorter spatial lag region. The derived theory was used to hypothesize and test previously unexplored principles for optimizing photoacoustic short-lag spatial coherence (SLSC) images, including the influence of the incident light profile on photoacoustic spatial coherence functions and associated SLSC image contrast and resolution. Results also confirm previous trends from experimental observations, including changes in SLSC image resolution and contrast as a function of the first M lags summed to create SLSC images. For example, small targets (e.g., <1-4 mm in diameter) can be imaged with larger M values to boost target contrast and resolution, and contrast can be further improved by reducing the illuminating beam to a size that is smaller than the target size. Overall, the presented theory provides a promising foundation to support a variety of coherence-based photoacoustic signal processing methods, and the associated theory-based simulation methods are more straightforward than the existing k-Wave simulation methods for SLSC images.

Simulations and human cadaver head studies to identify optimal acoustic receiver locations for minimally invasive photoacoustic-guided neurosurgery.

Real-time intraoperative guidance during minimally invasive neurosurgical procedures (e.g., endonasal transsphenoidal surgery) is often limited to endoscopy and CT-guided image navigation, which can be suboptimal at locating underlying blood vessels and nerves. Accidental damage to these critical structures can have severe surgical complications, including patient blindness and death. Photoacoustic image guidance was previously proposed as a method to prevent accidental injury. While the proposed technique remains promising, the original light delivery and sound reception components of this technology require alterations to make the technique suitable for patient use. This paper presents simulation and experimental studies performed with both an intact human skull (which was cleaned from tissue attachments) and a complete human cadaver head (with contents and surrounding tissue intact) in order to investigate optimal locations for ultrasound probe placement during photoacoustic imaging and to test the feasibility of a modified light delivery design. Volumetric x-ray CT images of the human skull were used to create k-Wave simulations of acoustic wave propagation within this cranial environment. Photoacoustic imaging of the internal carotid artery (ICA) was performed with this same skull. Optical fibers emitting 750 nm light were inserted into the nasal cavity for ICA illumination. The ultrasound probe was placed on three optimal regions identified by simulations: (1) nasal cavity, (2) ocular region, and (3) 1 mm-thick temporal bone (which received 9.2%, 4.7%, and 3.8% of the initial photoacoustic pressure, respectively, in simulations). For these three probe locations, the contrast of the ICA in comparative experimental photoacoustic images was 27 dB, 19 dB, and 12 dB, respectively, with delay-and-sum (DAS) beamforming and laser pulse energies of 3 mJ, 5 mJ, and 4.2 mJ, respectively. Short-lag spatial coherence (SLSC) beamforming improved the contrast of these DAS images by up to 15 dB, enabled visualization of multiple cross-sectional ICA views in a single image, and enabled the use of lower laser energies. Combined simulation and experimental results with the emptied skull and >1 mm-thick temporal bone indicated that the ocular and nasal regions were more optimal probe locations than the temporal ultrasound probe location. Results from both the same skull filled with ovine brains and eyes and the human cadaver head validate the ocular region as an optimal acoustic window for our current system setup, producing high-contrast (i.e., up to 35 dB) DAS and SLSC photoacoustic images within the laser safety limits of a novel, compact light delivery system design that is independent of surgical tools (i.e., a fiber bundle with 6.8 mm outer diameter, 2 mm-diameter optical aperture, and an air gap spacing between the sphenoid bone and fiber tips). These results are promising toward identifying, quantifying, and overcoming major system design barriers to proceed with future patient testing.

Additive noise models for photoacoustic spatial coherence theory.

Directly displaying the spatial coherence of photoacoustic signals (i.e., coherence-based photoacoustic imaging) remarkably improves image contrast, signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), and imaging depth when compared to conventional amplitude-based reconstruction techniques (e.g., backprojection, delay-and-sum beamforming, and Fourier-based reconstruction). We recently developed photoacoustic-specific theory to describe the spatial coherence process as a function of the element spacing on a receive acoustic aperture to enable photoacoustic image optimization without requiring experiments. However, this theory lacked noise models, which contributed to significant departures in coherence measurements when compared to experimental data, particularly at higher values of element separation. In this paper, we develop and implement two models based on experimental observations of noise in photoacoustic spatial coherence measurements to improve our existing spatial coherence theory. These models were derived to describe the effects of incident fluence variations, low-energy light sources (e.g., pulsed laser diodes and light-emitting diodes), averaging multiple signals from low-energy light sources, and imaging with light sources that are > 5mm from photoacoustic targets. Results qualitatively match experimental coherence functions and provide similar contrast, SNR, and CNR to experimental SLSC images. In particular, the added noise affects image quality metrics by introducing large variations in target contrast and significantly reducing target CNR and SNR when compared to minimal-noise cases. These results provide insight into additional requirements for optimization of coherence-based photoacoustic image quality.

Temporal Regulation by Innate Type 2 Cytokines in Food Allergies.

PURPOSE OF REVIEW: Food allergies (FAs) are a growing epidemic in western countries with poorly defined etiology. Defined as an adverse immune response to common food allergens, FAs present heterogeneously as a single- or multi-organ response that ranges in severity from localized hives and angioedema to systemic anaphylaxis. RECENT FINDINGS: Current research focusing on epithelial-derived cytokines contends that temporal regulation by these factors impact initial sensitization and persistence of FA responses upon repeated food allergen exposure. Mechanistic understanding of FA draws insight from a myriad of atopic conditions studied in humans and modeled in mice. In this review, we will highlight how epithelial-derived cytokines initiate and then potentiate FAs. We will also review existing evidence of the contribution of other atopic diseases to FA pathogenesis and whether FA symptoms overlap with other atopic diseases.

Label-Free Classification of Bax/Bak Expressing vs. Double-Knockout Cells.

We combine optical scatter imaging with principal component analysis (PCA) to classify apoptosis-competent Bax/Bak-expressing, and apoptosis resistant Bax/Bak-null immortalized baby mouse kidney cells. We apply PCA to 100 stacks each containing 236 dark-field cell images filtered with an optically implemented Gabor filter with period between 0.3 and 2.9 µm. Each stack yields an "eigencell" image corresponding to the first principal component obtained at one of the 100 Gabor filter periods used. At each filter period, each cell image is multiplied by (projected onto) the eigencell image. A Feature Matrix consisting of 236 × 100 scalar values is thus constructed with significantly reduced dimension compared to the initial dataset. Utilizing this Feature Matrix, we implement a supervised linear discriminant analysis and classify successfully the Bax/Bak-expressing and Bax/Bak-null cells with 94.7% accuracy and an area under the curve (AUC) of 0.993. Applying a feature selection algorithm further reveals that the Gabor filter period ranges most significant for the classification correspond to both large (likely nuclear) features as well as small sized features (likely organelles present in the cytoplasm). Our results suggest that cells with a genetic defect in their apoptosis pathway can be differentiated from their normal counterparts by label-free multi-parametric optical scatter data.

Structural basis of omalizumab therapy and omalizumab-mediated IgE exchange.

Omalizumab is a widely used therapeutic anti-IgE antibody. Here we report the crystal structure of the omalizumab-Fab in complex with an IgE-Fc fragment. This structure reveals the mechanism of omalizumab-mediated inhibition of IgE interactions with both high- and low-affinity IgE receptors, and explains why omalizumab selectively binds free IgE. The structure of the complex also provides mechanistic insight into a class of disruptive IgE inhibitors that accelerate the dissociation of the high-affinity IgE receptor from IgE. We use this structural data to generate a mutant IgE-Fc fragment that is resistant to omalizumab binding. Treatment with this omalizumab-resistant IgE-Fc fragment, in combination with omalizumab, promotes the exchange of cell-bound full-length IgE with omalizumab-resistant IgE-Fc fragments on human basophils. This combination treatment also blocks basophil activation more efficiently than either agent alone, providing a novel approach to probe regulatory mechanisms underlying IgE hypersensitivity with implications for therapeutic interventions.

Lessons learned from mice and man: mimicking human allergy through mouse models.

The relevance of using mouse models to represent human allergic pathologies is still unclear. Recent studies suggest the limitations of using models as a standard for assessing immune response and tolerance mechanisms, as mouse models often do not sufficiently depict human atopic conditions. Allergy is a combination of aberrant responses to innocuous environmental agents and the subsequent TH2-mediated inflammatory responses. In this review, we will discuss current paradigms of allergy - specifically, TH2-mediated and IgE-associated immune responses - and current mouse models used to recreate these TH2-mediated pathologies. Our overall goal is to highlight discrepancies that exist between mice and men by examining the advantages and disadvantages of allergic mouse models with respect to the human allergic condition.

Expression of the TEL-Syk fusion protein in hematopoietic stem cells leads to rapidly fatal myelofibrosis in mice.

The TEL-Syk fusion protein was isolated from a patient with myelodysplasia with megakaryocyte blasts. Expression of TEL-Syk transforms interleukin-3 (IL-3)-dependent Ba/F3 cells in vitro by deregulating STAT5-mediated signal transduction pathways. In vivo, TEL-Syk expression in pre-B cells blocks B cell differentiation, leading to lymphoid leukemia. Here, we demonstrate that TEL-Syk introduced into fetal liver hematopoietic cells, which are then adoptively transferred into lethally irradiated recipients, leads to an aggressive myelodysplasia with myelofibrosis that is lethal in mice by 60-75 days. Expression of TEL-Syk induces a short-lived myeloexpansion that is rapidly followed by bone marrow failure and extreme splenic/hepatic fibrosis accompanied by extensive apoptosis. The disease is dependent on Syk kinase activity. Analysis of serum from TEL-Syk mice reveals an inflammatory cytokine signature reminiscent of that found in the sera from patients and mouse models of myeloproliferative neoplasms. TEL-Syk expressing cells showed constitutive STAT5 phosphorylation, which was resistant to JAK inhibition, consistent with deregulated cytokine signaling. These data indicate that expression of TEL-Syk in fetal liver hematopoietic cells results in JAK-independent STAT5 phosphorylation ultimately leading to a uniquely aggressive and lethal form of myelofibrosis.