Brown adipose tissue evaluation using water and triglyceride as indices by diffuse reflectance spectroscopy.

Brown adipose tissue (BAT) is related to lipid and glucose metabolism, and BAT evaluation is expected to contribute to disease prevention and treatment. We aimed to establish a BAT evaluation method using simple and non-invasive diffuse reflectance spectroscopy (DRS). We acquired diffuse reflectance spectra of BAT using DRS from rats with cold stimulation and analyzed the second-derivative spectra. To predict the amount of triglyceride in BAT from the second-derivative spectra, partial least-squares regression analysis was performed, and we examined whether BAT weight can be predicted from the amount of triglyceride by single regression analysis. By focusing on changes in the amount of triglyceride in BAT with cold stimulation, it was suggested that this amount could be predicted spectroscopically, and the predicted amount of triglyceride could be used to estimate the BAT weight with cold stimulation. If these results can be translated into humans, they may contribute to preventing metabolic disorders.

Water and lipid content of breast tissue measured by six-wavelength time-domain diffuse optical spectroscopy.

Significance: The water and lipid content of normal breast tissue showed mammary gland characteristics with less influence from the chest wall using six-wavelength time-domain diffuse optical spectroscopy (TD-DOS) in a reflectance geometry. Aim: To determine the depth sensitivity of a six-wavelength TD-DOS system and evaluate whether the optical parameters in normal breast tissue can distinguish dense breasts from non-dense breasts. Approach: Measurements were performed in normal breast tissue of 37 breast cancer patients. We employed a six-wavelength TD-DOS system to measure the water and lipid content in addition to the hemoglobin concentration. The breast density in mammography and optical parameters were then compared. Results: The depth sensitivity of the system for water and lipid content was estimated to be ∼15 mm. Our findings suggest that the influence of the chest wall on the water content is weaker than that on the total hemoglobin concentration. In data with evaluation conditions, the water content was significantly higher (p < 0.001) and the lipid content was significantly lower (p < 0.001) in dense breast tissue. The water and lipid content exhibited a high sensitivity and specificity to distinguish dense from non-dense breasts in receiver-operating-characteristic curve analysis. Conclusions: With less influence from the chest wall, the water and lipid content of normal breast tissue measured by a reflectance six-wavelength TD-DOS system, together with ultrasonography, can be applied to distinguish dense from non-dense breasts.

Artificial intelligence-based classification of peripheral blood nucleated cells using label-free imaging flow cytometry.

Label-free image identification of circulating rare cells, such as circulating tumor cells within peripheral blood nucleated cells (PBNCs), the vast majority of which are white blood cells (WBCs), remains challenging. We previously described developing label-free image cytometry for classifying live cells using computer vision technology for pattern recognition, based on the subcellular structure of the quantitative phase microscopy images. We applied our image recognition methods to cells flowing in a flow cytometer microfluidic channel, and differentiated WBCs from cancer cell lines (area under receiver operating characteristic curve = 0.957). We then applied this method to healthy volunteers' and advanced cancer patients' blood samples and found that the non-WBC fraction rates (NWBC-FRs), defined as the percentage of cells classified as non-WBCs of the total PBNCs, were significantly higher in cancer patients than in healthy volunteers. Furthermore, we monitored NWBC-FRs over the therapeutic courses in cancer patients, which revealed the potential ability in monitoring the clinical status during therapy. Our image recognition system has the potential to provide a morphological diagnostic tool for circulating rare cells as non-WBC fractions.

Multiple-scattering suppressive refractive index tomography for the label-free quantitative assessment of multicellular spheroids.

Refractive index (RI) tomography is a quantitative tomographic technique used to visualize the intrinsic contrast of unlabeled biological samples. Conventional RI reconstruction algorithms are based on weak-scattering approximation, such as the Born or Rytov approximation. Although these linear algorithms are computationally efficient, they are invalid when the fields are strongly distorted by multiple scattering (MS) of specimens. Herein, we propose an approach to reconstruct the RI distributions of MS objects even under weak-scattering approximation using an MS-suppressive operation. The operation converts the distorted fields into MS-suppressed fields, where weak-scattering approximation is applicable. Using this approach, we reconstructed a whole multicellular spheroid and successfully visualized its internal subcellular structures. Our work facilitates the realization of RI tomography of MS specimens and label-free quantitative analysis of 3D multicellular specimens.

In vitro analysis of mechanism of pulsed-laser thrombolysis.

Thrombolytic therapy in the treatment of cardiogenic acute cerebral embolism caused by coagulated blood carries the risk of hemorrhagic complications, and there is a need to develop safer and more reliable treatment methods. Laser thrombolysis therapy, which utilizes the difference in energy absorption between the thrombus and the arterial wall, has shown promise as a new treatment method because it can selectively act only on the thrombus. It has not been applied clinically, however, and one of the main reasons for this is that its underlying mechanism has not been elucidated. We developed a pulse laser thrombolysis system for treating cerebral blood vessels that consists of a diode-pumped solid-state neodymium-yttrium aluminum garnet laser, which has excellent stability and maintainability and is suitable for clinical applications coupled to a small-diameter optical fiber. Moreover, we analyzed the mechanisms that occur during pulsed laser irradiation of transparent glass tubes and gelatin phantoms. We found that bubbles form as a thermal effect in addition to ablation of the pulsed laser irradiation. Furthermore, we detected no shock waves or water jets associated with the bubbles. We analyzed the bubbles' dynamics and growth rate, and their effect on a rabbit blood clot phantom. We concluded that the bubbles generated by the laser irradiation physically cut the thrombus and thereby had a thrombectomy effect. We believe that this study will clarify the mechanism of laser thrombolysis therapy and contribute greatly to the realization of its clinical application.

Single-shot quantitative phase imaging as an extension of differential interference contrast microscopy.

Various studies have been conducted to obtain quantitative phase information based on differential interference contrast (DIC) microscopy. As one such attempt, we propose in this study a single-shot quantitative phase imaging (QPI) method by combining two developments. First, an add-on optical system to a commercialized DIC microscope was developed to perform quantitative phase gradient imaging (QPGI) with single image acquisition using a polarization camera. Second, an algorithm was formulated to reconstitute QPI from the obtained QPGI by reducing linear artifacts, which arise in simply integrated QPGI images. To demonstrate the applicability of the developed system in cell biology, the system was used to measure various cell lines and compared with fluorescence microscopy images of the same field of view. Consistent with previous studies, nucleoli and lipid droplets can be imaged by the system with greater optical path lengths (OPL). The results also implied that combining fluorescence microscopy and the developed system might be more informative for cell biology research than using these methods individually. Exploiting the single-shot performance of the developed system, time-lapse imaging was also conducted to visualize the dynamics of intracellular granules in monocyte-/macrophage-like cells. Our proposed approach may accelerate the implementation of QPI in standard biomedical laboratories.

Label-free imaging flow cytometer for analyzing large cell populations by line-field quantitative phase microscopy with digital refocusing.

We propose a line-field quantitative phase-imaging flow cytometer for analyzing large populations of label-free cells. Hydrodynamical focusing brings cells into the focus plane of an optical system while diluting the cell suspension, resulting in decreased throughput rate. To overcome the trade-off between throughput rate and in-focus imaging, our cytometer involves digitally extending the depth-of-focus on loosely hydrodynamically focusing cell suspensions. The cells outside the depth-of-focus range in the 70-µm diameter of the core flow were automatically digitally refocused after image acquisition. We verified that refocusing was successful with our cytometer through statistical analysis of image quality before and after digital refocusing.

Effect of Source-Detector Distance on the Measurement of Hemoglobin Using Near-Infrared Spectroscopy in Breast Cancer.

We measured total hemoglobin concentrations in breast tumors by near-infrared time-resolved spectroscopy. Muscles interfere with measurement when the probe is close to the chest wall. Since the target area of measurement depends on the distance between the light source and probe detector, we inferred that this issue could be solved by reducing the source-detector distance. The purpose of this study was to examine the effects of the source-detector distance on the measurement of total hemoglobin concentration in the breast. We examined 26 patients with breast tumors. Total hemoglobin concentration was measured in tumors and the contralateral normal breasts at source-detector distances of 20 and 30 mm. The difference in total hemoglobin concentration between each tumor and the contralateral breast was calculated. The normal breast total hemoglobin concentration was significantly smaller for the source-detector distance of 20 mm than for the source-detector distance of 30 mm. Differences in source-detector distance did not significantly affect tumor total hemoglobin. The difference in total hemoglobin concentration between the tumor and the contralateral breast obtained at the source-detector distance of 20 mm was significantly higher than that obtained at the source-detector distance of 30 mm. From these results, we considered that measurement with a source-detector distance of 20 mm is less affected by the chest wall than with a source-detector distance of 30 mm and that the difference in total hemoglobin concentration between the tumor and the contralateral breast at a source-detector distance of 20 mm can better reflect the net total hemoglobin concentrations of the breast tumors. In conclusion, using a probe with a source-detector distance of 20 mm can more accurately evaluate the total hemoglobin concentration in breast tumors.

Label-free classification of cells based on supervised machine learning of subcellular structures.

It is demonstrated that cells can be classified by pattern recognition of the subcellular structure of non-stained live cells, and the pattern recognition was performed by machine learning. Human white blood cells and five types of cancer cell lines were imaged by quantitative phase microscopy, which provides morphological information without staining quantitatively in terms of optical thickness of cells. Subcellular features were then extracted from the obtained images as training data sets for the machine learning. The built classifier successfully classified WBCs from cell lines (area under ROC curve = 0.996). This label-free, non-cytotoxic cell classification based on the subcellular structure of QPM images has the potential to serve as an automated diagnosis of single cells.

Stable tissue-simulating phantoms with various water and lipid contents for diffuse optical spectroscopy.

We introduced a method for producing solid phantoms with various water-to-lipid ratios that can simulate the absorption, and to some extent the scattering characteristics of human breast tissue. We also achieved phantom stability for a minimum of one month by solidifying the emulsion phantoms. The characteristics of the phantoms were evaluated using the six-wavelength time-domain diffuse optical spectroscopy (TD-DOS) system we developed to measure water and lipid contents and hemoglobin concentration. The TD-DOS measurements were validated with a magnetic resonance imaging system.

Monitoring of Brain Oxygenation During and After Cardiopulmonary Resuscitation: A Prospective Porcine Study.

This study aimed to evaluate the usefulness of near-infrared time-resolved spectroscopy (TRS) for the monitoring of post-resuscitation encephalopathy. Cardiac arrest (CA) was induced in pigs by electrical stimuli; then, return of spontaneous circulation (ROSC) was achieved by direct current. The changes in cerebral oxygenation were analyzed by two methods: (1) the time-independent calculation based on the modified Beer-Lambert law (MBL), and (2) the curve-fitting method based on the photon diffusion theory (DT). The changes in reduced scattering coefficient (µs') in DT were also calculated. Post-resuscitation encephalopathy was evaluated by MRI findings. During CA, cerebral oxygen saturation (ScO2) decreased to the lowest level, and then gradually increased during the chest compression period. When ROSC was achieved, ScO2 (DT) increased further, but ScO2 (MBL) decreased transiently. This strange phenomenon disappeared when the scalp was peeled off and the probes were directly fixed to the cranial bone. In some cases, a sustained decrease in µs' was observed several hours after ROSC and, in such cases, MRI Diffusion Enhancement Image (DWI) showed findings suggestive of post-resuscitation encephalopathy. In conclusion, simultaneous monitoring of cerebral oxygenation with MBL and DT may provide more information about the vascular response of different layers. Also, the monitoring of µs' may help us to recognize the occurrence of post-resuscitation encephalopathy in real time.

Fracture probability assessed using FRAX® in elderly women with benign paroxysmal positional vertigo.

OBJECTIVE: Patients with benign paroxysmal positional vertigo (BPPV) can have vitamin D deficiency, which is a cause of abnormal bone turnover. Several studies have established a relationship between osteoporosis and BPPV. The World Health Organization Fracture Risk Assessment Tool, widely known as FRAX® (http://www.shef.ac.uk/FRAX), is a computer-based algorithm for assessing fracture risk. No direct comparison has been made between the FRAX scores of patients with BPPV and controls. The purpose of this study was to determine whether women with BPPV are at high risk of fracture as assessed using FRAX. METHODS: The study involved 40 postmenopausal women diagnosed with BPPV between July 2015 and April 2016, and 40 postmenopausal women as controls. The 10-year major osteoporotic and hip fracture risks were calculated using FRAX and were compared between BPPV patients and controls using Welch's t test and a general linear model. RESULTS: The 10-year major osteoporotic fracture risk was 20.4%±12.1% for BPPV patients (aged 72.4±8.6years) and 14.3%±6.5% for controls (aged 71.2±6.3years). The 10-year hip fracture risk was 9.0%±9.8% for BPPV patients and 5.0%±3.9% for controls. The BPPV group had significantly higher 10-year major risks of osteoporotic fracture (p=0.0069) and hip fracture (p=0.0202) compared with controls. Similarly, after adjustment for age, the BPPV group had significantly higher 10-year risks of major osteoporotic fracture (p=0.0007) and hip fracture (p=0.0092) compared with controls. CONCLUSION: Fracture risk calculated using FRAX was significantly higher in the BPPV group than in controls. Women with BPPV may need early intervention to prevent future fractures.

Factors affecting measurement of optic parameters by time-resolved near-infrared spectroscopy in breast cancer.

The purpose of this study was to evaluate the effects of the thickness and depth of tumors on hemoglobin measurements in breast cancer by optical spectroscopy and to demonstrate tissue oxygen saturation (SO2) and reduced scattering coefficient (µs') in breast tissue and breast cancer in relation to the skin-to-chest wall distance. We examined 53 tumors from 44 patients. Total hemoglobin concentration (tHb), SO2, and µs' were measured by time-resolved spectroscopy (TRS). The skin-to-chest wall distance and the size and depth of tumors were measured by ultrasonography. There was a positive correlation between tHb and tumor thickness, and a negative correlation between tHb and tumor depth. SO2 in breast tissue decreased when the skin-to-chest wall distance decreased, and SO2 in tumors tended to be lower than in breast tissue. In breast tissue, there was a negative correlation between µs' and the skin-to-chest wall distance, and µs' in tumors was higher than in breast tissue. Measurement of tHb in breast cancer by TRS was influenced by tumor thickness and depth. Although SO2 seemed lower and µs' was higher in breast cancer than in breast tissue, the skin-to-chest wall distance may have affected the measurements.

Near-Infrared Diffuse Optical Imaging for Early Prediction of Breast Cancer Response to Neoadjuvant Chemotherapy: A Comparative Study Using 18F-FDG PET/CT.

UNLABELLED: Diffuse optical spectroscopic imaging (DOSI) is used as an indicator of tumor blood volume quantified by tissue hemoglobin concentrations. We aimed to determine whether early changes in tumor total hemoglobin (tHb) concentration can predict a pathologic complete response (pCR) to neoadjuvant chemotherapy in patients with operable breast cancer, and we compared the predictive value of pCR between DOSI and (18)F-FDG PET combined with CT. METHODS: Of the 100 patients enrolled, 84 patients were prospectively evaluated for primary objective analysis. Sixty-four of the patients underwent both sequential DOSI scans at baseline after their first and second chemotherapy courses and (18)F-FDG PET/CT at baseline and after their second chemotherapy course. The mean tHb (tHbmean) concentration and SUVmax of the lesion were measured using DOSI and (18)F-FDG PET/CT, respectively, and the percentage change in tHbmean (∆tHbmean) and change in SUVmax (∆SUVmax) were calculated. We compared the diagnostic performances of DOSI and (18)F-FDG PET/CT for predicting pCR via the analysis of the receiver-operating-characteristic curves. RESULTS: pCR was achieved in 16 patients, and neoadjuvant chemotherapy caused a significant reduction of ∆tHbmean in pCR compared with non-pCR after the 2 chemotherapy courses. When the tentative ∆tHbmean cutoff values after the first and second courses were used, the ability to predict pCR was as follows: 81.2% sensitivity/47.0% specificity and 93.7% sensitivity/47.7% specificity, respectively. Comparison of the diagnostic performances of DOSI and (18)F-FDG PET/CT revealed areas under the curve of 0.69 and 0.75 of ∆tHbmean after the first and second courses, respectively, which were lower than those of ∆SUVmax (0.90). CONCLUSION: DOSI predicted pCR in patients with breast cancer with moderate accuracy. The diagnostic performance of DOSI was inferior to that of the early metabolic response as monitored by (18)F-FDG PET/CT.

Effect of the chest wall on the measurement of hemoglobin concentrations by near-infrared time-resolved spectroscopy in normal breast and cancer.

BACKGROUND: Optical imaging and spectroscopy using near-infrared light have great potential in the assessment of tumor vasculature. We previously measured hemoglobin concentrations in breast cancer using a near-infrared time-resolved spectroscopy system. The purpose of the present study was to evaluate the effect of the chest wall on the measurement of hemoglobin concentrations in normal breast tissue and cancer. METHODS: We measured total hemoglobin (tHb) concentration in both cancer and contralateral normal breast using a near-infrared time-resolved spectroscopy system in 24 female patients with breast cancer. Patients were divided into two groups based on menopausal state. The skin-to-chest wall distance was determined using ultrasound images obtained with an ultrasound probe attached to the spectroscopy probe. RESULTS: The apparent tHb concentration of normal breast increased when the skin-to-chest wall distance was less than 20 mm. The tHb concentration in pre-menopausal patients was higher than that in post-menopausal patients. Although the concentration of tHb in cancer tissue was statistically higher than that in normal breast, the contralateral normal breast showed higher tHb concentration than cancer in 9 of 46 datasets. When the curves of tHb concentrations as a function of the skin-to-chest wall distance in normal breast were applied for pre- and post-menopausal patients separately, all the cancer lesions plotted above the curves. CONCLUSIONS: The skin-to-chest wall distance affected the measurement of tHb concentration of breast tissue by near-infrared time-resolved spectroscopy. The tHb concentration of breast cancer tissue was more precisely evaluated by considering the skin-to-chest wall distance.

Reduction of moiré for coarse integral volumetric imaging.

This paper proposes two moiré reduction methods for coarse integral volumetric displays composed of multilayer display panels. The quality of the image presented by the conventional multilayer integral imaging stays relatively low because of the moiré pattern, which appears due to the layering structure of display panels with limited pixel aperture. The authors propose two methods to solve this problem. The first method is to add another layer of convex lens array with short focal length on the front side of the layered display panels. The second method is to enlarge the pixel pitch of the back-layer panel while fixing that of the front-layer panel.

Integral volumetric imaging with dual layer fly-eye lenses.

This paper proposes an integral volumetric imaging system that uses a coarse fly-eye lens and a fine fly-eye lens to show smooth and deep 3D image. Conventional integral volumetric imaging displays using a coarse fly-eye lens have suffered from low image quality due to distinct seam of lenses and moiré pattern caused by layered panel structure. To solve these problems the proposed system uses a fine fly-eye lens whose elemental lens has a long focal distance. By placing a fine fly-eye lens near the layered real image, the seam and the moiré are removed while the degradation of the presented 3D image is kept small.

Low-density lipoprotein apheresis therapy with a direct hemoperfusion column: a Japanese multicenter clinical trial.

Low-density lipoprotein (LDL) apheresis has been applied to patients with familial hypercholesterolemia (FH) with coronary artery disease (CAD). To examine the efficacy and safety of a new type of LDL adsorption column (KLD01, Kaneka, Osaka, Japan), which deals with whole blood without separating plasma, the new system was evaluated in a multicenter trial. The present study included 33 FH patients with CAD (24 males, 9 females, 57 +/- 13 years) who were treated five times with a mean interval of 2.12 +/- 0.60 weeks between treatments. We studied the removal efficacies for serum LDL cholesterol, Lipoprotein(a) (Lp(a)) and triglyceride, the times for the preparation of the system and for treatment, symptoms, and the biochemical data. The scheduled treatments were completed by 31 patients. Serum levels of LDL cholesterol, Lp(a) and triglycerides were all significantly reduced with KLD01; 61.5 +/- 6.2%, 72.4 +/- 5.9% and 69.5 +/- 9.7%, respectively. The times for both setting up the column system (26 +/- 7 min) and treatment (138 +/- 20 min) were shorter with KLD01 than conventional methods. Adverse reactions occurred in eight cases (17 episodes), but the patients fully recovered immediately after each apheresis therapy session. We conclude that the new type of LDL adsorption column, one that deals with whole blood, is a promising apheresis therapy for FH patients in view of its efficacy, reduced time for treatment, and safety.