3-epi-18ß-glycyrrhetinic acid or its glucuronide, the metabolites of glycyrrhizinic acid with individual differences, correlated with diagnostic maker for licorice-induced pseudoaldosteronism in humans.

Licorice is a crude drug that is used in traditional Japanese Kampo medicine and is also used as a sweetener. Occasionally, it causes pseudoaldosteronism (PsA) as a side effect. The major symptoms include hypokalemia, hypertension, edema, and low plasma aldosterone levels. PsA might be caused by the metabolites of glycyrrhizinic acid (GL), a component of licorice. The development of PsA markedly varies among individuals; however, the factors that cause these individual differences remain unknown. In this study, 78 patients who consumed Kampo medicines containing licorice were enrolled, and their laboratory data, including serum potassium levels, plasma aldosterone concentrations (PAC), and the concentrations of GL metabolites in the residual blood and/or urine samples were evaluated. Of the 78 participants, 18ß-glycyrrhetinic acid (GA), 3-epi-GA, 3-oxo-GA, 18ß-glycyrrhetinyl-30-O-glucuronide (GA30G), and 3-epi-GA30G were detected in the serum samples of 65, 47, 63, 62, and 3 participants, respectively. Of the 29 urine samples collected, GA30G and 3-epi-GA30G were detected in 27 and 19 samples. 3-epi-GA30G is a newly found GL metabolite. Moreover, 3-epi-GA, 3-oxo-GA, and 3-epi-GA30G were identified in human samples for the first time. High individual differences were found in the appearances of 3-epi-GA in serum and 3-epi-GA30G in urine, and the concentrations of these metabolites were correlated with serum PsA markers. The inhibitory titers of 3-epi-GA, 3-oxo-GA, GA30G, and 3-epi-GA30G on human 11ß-hydroxysteroid dehydrogenase type 2 (11ß-HSD2) were almost similar. These findings suggest that 3-epi-GA and/or 3-epi-GA30G are associated with individual differences in the development of PsA. Significance Statement In this study, we detected 3-epi-GA in human serum for the first time. We also identified 3-epi-GA30G as a novel GL metabolite in human urine. These GL metabolite levels showed correlations with markers of PsA. Additionally, there are individual differences in whether or not they appear in the serum/urine. In conclusion, 3-epi-GA/3-epi-GA30G correlates with individual differences in the development of PsA.

Development of telemedicine tools with an emphasis on visual observation.

We developed a system to improve the quality of telemedicine, and the test results obtained have been presented in this paper, along with the technical details of the system. The spread of COVID-19 has accelerated the need for telemedicine to effectively prevent infections. However, in traditional Japanese medicine (Kampo), where color is essential, an accurate diagnosis cannot be made without color reproduction. Because commercial smartphones cannot reproduce colors with the level of fidelity required for medical treatments, we created a color chart that includes the human skin and tongue colors to help doctors identify their colors accurately during a telemedicine examination. Further, we developed a telemedicine system that allows for automatic color correction using a mobile device, with a color chart and non-contact heart rate measurements.

Assessing the effects of Kampo medicine on human skin texture and microcirculation.

In this study, we verified the effectiveness of Kampo medicine by evaluating the changes in the feature values of facial skin texture and microcirculation at two distinct tissue depths (subcutaneous 2 mm and 8 mm). A total of 80 patients who took the Kampo formula participated in this study, and the changes in the feature values of facial skin texture and microcirculation were measured before and after Kampo treatment. The treatment period lasted 6-18 months, according to the doctor's judgment. The total area of the sulci cutis and the average thickness of the sulci cutis significantly decreased (P < 0.05), and the pixels of the grayscale image increased after Kampo treatment (P < 0.05). Moreover, the blood flow velocity at 8 mm depth significantly increased after Kampo treatment (P < 0.05). In this study, we specifically noted changes in the skin texture and microcirculation after Kampo treatment.

Development of a camera-based remote diagnostic system focused on color reproduction using color charts.

In this paper, we propose a color reproduction method using color charts to improve the color quality of a telemedicine system. Owing to the spread of COVID-19, the need for telemedicine is rapidly increasing to prevent infections more effectively. However, in practices such as traditional Japanese (Kampo) medicine, where color is used as an important examination factor, an accurate diagnosis cannot be made without adequate color reproduction. In telemedicine using a commercially available smartphone, color reproducibility may deteriorate owing to differences in the devices and lighting, which may result in a misdiagnosis. Therefore, we created a color chart that includes the colors of the human skin and tongue as a tool to help doctors identify the color of patients more accurately when conducting a telemedicine examination. Through a subjective evaluation by eight medical doctors, it was unanimously found that the proposed method is practical in terms of a color examination. The developed color chart can also be used for an automatic color correction.

Identification of glycyrrhizin metabolites in humans and of a potential biomarker of liquorice-induced pseudoaldosteronism: a multi-centre cross-sectional study.

Liquorice [main ingredient, glycyrrhizin (GL)] is widely used as a food sweetener and herbal medicine. Occasionally, liquorice consumption causes pseudoaldosteronism as a side effect which causes oedema, hypokalaemia, and hypertension due to hyperactivity of mineral corticoid receptor. We aimed to detect GL metabolites in human blood and urine samples and to determine the pathological relationship between GL metabolites and pseudoaldosteronism. For this multi-centre, retrospective, cross-sectional study, we recruited patients who had visited Center for Kampo Medicine in Keio University Hospital, Department of Japanese Oriental (Kampo) Medicine in Chiba University Hospital, Clinic of Japanese Oriental (Kampo) Medicine in Kanazawa University Hospital, and Department of Oriental Medicine in Kameda Medical Center from November 2011 to July 2018. We collected laboratory data including concentration of serum potassium, plasma activity of renin and aldosterone, and residual blood and/or urine samples of participants who had experienced symptoms/signs of pseudoaldosteronism in the form of increase in blood pressure and occurrence or aggregation of oedema while taking liquorice-containing herbal preparations, and measured GL metabolites using a highly selective liquid chromatography tandem mass spectrometer system. We registered 97 participants (mean age 60 ± 15 years; male:female 14:83). 18ß-glycyrrhetinic acid (GA) was detected in 67 serum samples (median 122 nM, range 5 nM-1.8 µM) and 18ß-glycyrrhetyl-3-O-sulfate (compound 3) in 68 samples (median 239 nM, range 2 nM-4.2 µM). 3-Monoglucuronyl 18ß-glycyrrhetinic acid, 22α-hydroxy-18ß-glycyrrhetyl-3-O-sulfate-30-glucuronide, 22α-hydroxy-18ß-glycyrrhetyl-3-O-sulfate, and GL itself were not or rarely detected. We could not find any correlation between blood pressure or peripheral oedema and serum concentration of GL metabolites. Sulfotransferase 2A1 catalysed the metabolic reaction of GA to compound 3, a major GL metabolite in human blood. High serum concentration of compound 3 was related to lower renin, aldosterone, and potassium levels, suggesting a pathological relationship between compound 3 and liquorice-induced pseudoaldosteronism. This is the first study to identify the association between a novel metabolite, compound 3, and the incidence of pseudoaldosteronism, highlighting it as a promising biomarker.

Quantitative Capillary Refill Time with image-based finger force estimation.

Skin color observation provides a simple and non-invasive method to estimate the health status of patients. Capillary Refill Time (CRT) is widely used as an indicator of pathophysiological conditions, especially in emergency patients. While the measurement of CRT is easy to perform, its evaluation is highly subjective. This study proposes a method to aid quantified CRT measurement using an RGB camera. The procedure consists in applying finger compression to the forearm, and the CRT is calculated based on the skin color change after the pressure release. We estimate compression applied by a finger from its fingernail color change during compression. Our study shows a step towards camera-based quantitative CRT for untrained individuals.

Beneficial Effects of Ginger Extract on Eye Fatigue and Shoulder Stiffness: A Randomized, Double-Blind, and Placebo-Controlled Parallel Study.

This study aimed to assess ginger extract's impact on ocular and peripheral blood flow and its potential to alleviate eye fatigue and shoulder stiffness. This study included 100 healthy individuals aged 20-73 years with eye fatigue and shoulder stiffness. Participants were randomly assigned to receive either placebo capsules or ginger extract capsules daily for eight weeks. Ocular blood flow, peripheral blood flow, eye fatigue (visual analog scale [VAS]), shoulder stiffness (VAS), body warmth (VAS), and shoulder muscle stiffness were assessed at weeks 0, 4, and 8, respectively. No improvement in ocular blood flow was observed under the study conditions. Conversely, peripheral blood flow in deep areas was enhanced in females (p = 0.033). Subgroup analysis by age (≥51 or <51 years) revealed that ginger's effect on enhancing peripheral blood flow in deep vessels was restricted in females under 51 (p = 0.017). Similarly, subjective complaints of eye fatigue and shoulder stiffness were improved by ginger consumption in females under 51. Body warmth was favorably changed significantly in males ≥51 years due to ginger consumption. The muscle stiffness showed no statistically significant changes. In conclusion, ginger consumption reduces eye fatigue and shoulder stiffness by enhancing peripheral blood flow in relatively young females.

Noise-Robust Pulse Wave Estimation from Near-Infrared Face Video Images Using the Wiener Estimation Method.

In this paper, we propose a noise-robust pulse wave estimation method from near-infrared face video images. Pulse wave estimation in a near-infrared environment is expected to be applied to non-contact monitoring in dark areas. The conventional method cannot consider noise when performing estimation. As a result, the accuracy of pulse wave estimation in noisy environments is not very high. This may adversely affect the accuracy of heart rate data and other data obtained from pulse wave signals. Therefore, the objective of this study is to perform pulse wave estimation robust to noise. The Wiener estimation method, which is a simple linear computation that can consider noise, was used in this study. Experimental results showed that the combination of the proposed method and signal processing (detrending and bandpass filtering) increased the SNR (signal to noise ratio) by more than 2.5 dB compared to the conventional method and signal processing. The correlation coefficient between the pulse wave signal measured using a pulse wave meter and the estimated pulse wave signal was 0.30 larger on average for the proposed method. Furthermore, the AER (absolute error rate) between the heart rate measured with the pulse wave meter was 0.82% on average for the proposed method, which was lower than the value of the conventional method (12.53% on average). These results show that the proposed method is more robust to noise than the conventional method for pulse wave estimation.

Preliminary research of the effectiveness of acupuncture (spoon needle) on stress relief of Japanese expatriates in a developing country Myanmar: A retrospective study.

This was a preliminary retrospective study to clarify the effectiveness of traditional Japanese acupuncture on the stress level of Japanese expatriates who studied or worked in Myanmar. Total 22 Japanese expatriate patients were selected for this study. The profile of mood states second edition-adult short form scale was used to evaluate the stress-relief effect of acupuncture before every treatment. Negative mood (tension and confusion) and total mood disturbances significantly decreased after acupuncture treatment. Regarding the influence of background, the experience of staying abroad significantly influenced the efficacy of the acupuncture treatment. These results indicate that acupuncture treatment can relieve stress experienced by Japanese expatriates. The results also suggest that the experience of staying abroad might be one of the leading factors that can modulate the effects of treatment.

Blood flow in the optic nerve head in patients with primary aldosteronism.

PURPOSE: Optic nerve head (ONH) blood flow decrease without changes in intraocular pressure in a possible rat model of retinal ganglion cell loss by systemic administration of aldosterone. To compare the blood flow in the ONH, using laser speckle flowgraphy (LSFG), in healthy eyes and in eyes with primary aldosteronism (PA). METHODS: The ONH tissue area mean blur rate (MT) was evaluated in this single center, retrospective, cross-sectional study using LSFG. In order to compare the MT between PA patients and normal subjects, mixed-effects models were used, with adjustments made for the mean arterial pressure, disc area, and ß-peripapillary atrophy (ß-PPA) area. Mixed-effects models were also used to analyze the risk factors affecting the MT. RESULTS: This study evaluated a total of 29 eyes of 17 PA patients and 61 eyes of 61 normal subjects. There was a significantly lower MT in PA patients (10.8 ± 0.4) as compared to the normal subjects (12.3 ± 0.3) (P = 0.004). The MT was significantly lower in PA patients (10.8 ± 0.6) even after adjusting for the potential confounding factors when compared to normal subjects (12.3 ± 0.3) (P = 0.046). Multivariate mixed-effects model analysis demonstrated that the MT was significantly associated with the PA and ß-PPA. CONCLUSIONS: There was a significantly lower ONH blood flow in PA patients as compared to normal subjects.

Efficiency of Japanese herbal medicine shokenchuto for nocturnal enuresis: An observational study.

Japanese traditional (Kampo) medicine has been empirically used for nocturnal enuresis (NE). This study aims to investigate the efficacy of one of the most popular formulas, shokenchuto (SKT). We retrospectively analyzed 110 patients with NE who were referred to our department. Following the diagnosis of NE, treatment was started with either alarm or/and desmopressin (DDAVP) therapy. Patient refractory to DDAVP monotherapy or to combination therapy consisting of DDAVP and bedwetting alarm were selected. SKT (Tsumura Co., Tokyo, Japan) extract at a dose of 2.5 g was administered orally to all intractable cases twice daily before meals. The treatment outcomes and safety were assessed. In total, 24 cases were patient refractory to DDAVP monotherapy or to combination therapy consisting of DDAVP and bedwetting alarm. SKT was highly effective in 8, effective in 7, and ineffective in 9. A significant difference was observed between ages 10 and over (P = 0.031). SKT was significantly effective as a treatment for NE in patients aged ≥10 years and could be a good alternative if alarm or DDAVP therapies are ineffective. We proposed evaluating SKT prospectively for NE.

Blood pressure estimation by spatial pulse-wave dynamics in a facial video.

We propose a remote method to estimate continuous blood pressure (BP) based on spatial information of a pulse-wave as a function of time. By setting regions of interest to cover a face in a mutually exclusive and collectively exhaustive manner, RGB facial video is converted into a spatial pulse-wave signal. The spatial pulse-wave signal is converted into spatial signals of contours of each segmented pulse beat and relationships of each segmented pulse beat. The spatial signal is represented as a time-continuous value based on a representation of a pulse contour in a time axis and a phase axis and an interpolation along with the time axis. A relationship between the spatial signals and BP is modeled by a convolutional neural network. A dataset was built to demonstrate the effectiveness of the proposed method. The dataset consists of continuous BP and facial RGB videos of ten healthy volunteers. The results show an adequate estimation of the performance of the proposed method when compared to the ground truth in mean BP, in both the correlation coefficient (0.85) and mean absolute error (5.4 mmHg). For comparison, the dataset was processed using conventional pulse features, and the estimation error produced by our method was significantly lower. To visualize the root source of the BP signals used by our method, we have visualized spatial-wise and channel-wise contributions to the estimation by the deep learning model. The result suggests the spatial-wise contribution pattern depends on the blood pressure, while the pattern of pulse contour-wise contribution pattern reflects the relationship between percussion wave and dicrotic wave.

Clinical and epidemiological features of healthcare workers after a coronavirus disease 2019 cluster infection in Japan and the effects of Kampo formulas-Hochuekkito and Kakkonto: A retrospective cohort study.

It is expected that a low-toxicity natural compound like Kampo formulas would exhibit a preventive effect on COVID-19, in a global outbreak of coronavirus disease 2019 (COVID-19). Although the biological properties and safety of the representative Kampo, Hochuekkito (HET), and Kakkonto (KKT) have been confirmed in various animal model experiments and clinical studies, and in a few human studies to induce biological effects on various infectious diseases without significant toxicity, it is unclear whether HET and KKT are safe and effective for COVID-19 prevention. We summarized the clinical characteristics of HCWs and the preventive effects of HET and KKT. We performed a retrospective, single-center, cohort study that included 175 HCWs (aged 21-77 years) from a total number of 217 in a hospital with a history of COVID-19 cluster infection. In total, 175 HCWs were tested for severe acute respiratory coronavirus virus 2 (SARS-CoV-2) antibodies. We identified 27 patients (median age: 49 ± 10.7 years) who were diagnosed with COVID-19. The patients in the group that had a body mass index ≥ 25 had a high COVID-19 infection risk, while those in the group with a Kampo formula adherence rate ≥ 40% had a low COVID-19 risk. Patients in the group with an adherence rate ≥ 40%, as well as those in the current alcohol consumption group, were at a low risk of developing severe COVID-19. In conclusion, HET and KKT may have prevented the onset or worsening of COVID-19, which could be clinically used. Obesity might have increased the patients' susceptibility to COVID-19 and the disease severity.

U-Net-Based Segmentation of Microscopic Images of Colorants and Simplification of Labeling in the Learning Process.

Colored product textures correspond to particle size distributions. The microscopic images of colorants must be divided into regions to determine the particle size distribution. The conventional method used for this process involves manually dividing images into areas, which may be inefficient. In this paper, we have overcome this issue by developing two different modified architectures of U-Net convolution neural networks to automatically determine the particle sizes. To develop these modified architectures, a significant amount of ground truth data must be prepared to train the U-Net, which is difficult for big data as the labeling is performed manually. Therefore, we also aim to reduce this process by using incomplete labeling data. The first objective of this study is to determine the accuracy of our modified U-Net architectures for this type of image. The second objective is to reduce the difficulty of preparing the ground truth data by testing the accuracy of training on incomplete labeling data. The results indicate that efficient segmentation can be realized using our modified U-Net architectures, and the generation of ground truth data can be simplified. This paper presents a preliminary study to improve the efficiency of determining particle size distributions with incomplete labeling data.

In vitro Suppression of SARS-CoV-2 Infection by Existing Kampo Formulas and Crude Constituent Drugs Used for Treatment of Common Cold Respiratory Symptoms.

Several traditional Japanese Kampo formulas are known to have inhibitory effects on infections with viruses that cause respiratory symptoms. Although some herbs and their components have been reported to suppress SARS-CoV-2 replication in vitro, it is difficult to compare effective Kampo formulas because of the different methods used in studies. Thus, we carried out in vitro experiments on the suppression of SARS-CoV-2 infection by Kampo formulas and crude drugs used for the common cold to compare their suppressive effects on virus infection. After infecting VeroE6/TMPRSS2 cells with SARS-CoV-2, lysates of the Kampo formulas and crude drugs were added, and after 24 h, the infectious titer in the medium was measured by the TCID50 method. Maoto was the most effective among the Kampo formulas, and Ephedrae herba was the most effective among the constituent crude drugs. However, a comparison of the suppressive effects of Ephedrae herba and Kampo formulas containing Ephedrae herba showed that the suppressive effect on virus infection did not depend on the content of Ephedrae herba. Based on the results, we believe that the use of Maoto among Kampo formulas is suitable as a countermeasure against COVID-19.

Corrigendum: Identification of an alternative glycyrrhizin metabolite causing liquorice-induced pseudohyperaldosteronism and the development of ELISA system to detect the predictive biomarker.

[This corrects the article DOI: 10.3389/fphar.2021.688508.].

A Circumference-Measurement Method Using a Model of a Leg and a 3D Camera.

The circumference of a limb is an important parameter in the follow-up of an edema. Recently, several methods of measuring the circumference on a limb using 3D cameras have been proposed. However, the 3D cameras used are expensive and difficult to implement in general medical facilities. In this study, we propose a circumference-measurement method using a Structure Sensor. First, the leg is photographed and unnecessary background objects are removed from the obtained point cloud. Next, a cross-sectional view is obtained by slicing the point cloud at the specified leg height. Finally, the circumference measurement at a specified leg height is performed by calculating the circumference using the acquired cross-sectional view. Using this method, the leg circumferences of two healthy subjects were measured at two points. For comparison, circumferences were also measured with a measuring tape. The difference between the values estimated using our method and the measured values was generally less than 0.5 cm.

Effect of Japanese Kampo medicine, eppikajutsuto, in patients with lymphatic malformation: A retrospective observational study.

ABSTRACT: Lymphatic malformations (LMs) are congenital malformations of the lymphatic system that cause considerable cosmetic and functional complications. In this study, we present 8 children with LM who were treated with the Kampo medicine eppikajutsuto (EKJT).Between 2001 and 2020, 8 children (male: 4, female: 4) with LMs who underwent magnetic resonance imaging (MRI) evaluation both before and after treatment or observation were selected for investigating the effect of EKJT. Two patients were observed without any treatment for 24 and 60 months. EKJT was evaluated based on percentage reduction, defined as the percentage of total lesions that decreased in size, confirmed by radiological examination after initiating treatment with EKJT or determined by observation alone. Volumetric analysis of LMs on MRI was performed using the Digital Imaging and Communications in Medicine viewer.Six patients were treated with EKJT. The mean observational period was 13.2 months (range: 6-24 months). The mean reduction in LM volume on MRI was 73.0% in treated patients and -66.3% in observed patients. Two of the 6 lesions exhibited complete reduction, 2 exhibited marked (>90%) reduction, 1 exhibited moderate reduction, and 1 exhibited a small response. The treatment was well-tolerated, with no severe adverse events.This preliminary study demonstrated the beneficial effects of EKJT. Prospective evaluations of this promising therapeutic modality are warranted based on the results of this study.

Glial cell type-specific gene expression in the mouse cerebrum using the piggyBac system and in utero electroporation.

Glial cells such as astrocytes and oligodendrocytes play crucial roles in the central nervous system. To investigate the molecular mechanisms underlying the development and the biological functions of glial cells, simple and rapid techniques for glial cell-specific genetic manipulation in the mouse cerebrum would be valuable. Here we uncovered that the Gfa2 promoter is suitable for selective gene expression in astrocytes when used with the piggyBac system and in utero electroporation. In contrast, the Blbp promoter, which has been used to induce astrocyte-specific gene expression in transgenic mice, did not result in astrocyte-specific gene expression. We also identified the Plp1 and Mbp promoters could be used with the piggyBac system and in utero electroporation to induce selective gene expression in oligodendrocytes. Furthermore, using our technique, neuron-astrocyte or neuron-oligodendrocyte interactions can be visualized by labeling neurons, astrocytes and oligodendrocytes differentially. Our study provides a fundamental basis for specific transgene expression in astrocytes and/or oligodendrocytes in the mouse cerebrum.

Identification of an Alternative Glycyrrhizin Metabolite Causing Liquorice-Induced Pseudohyperaldosteronism and the Development of ELISA System to Detect the Predictive Biomarker.

Liquorice is usually used as crude drug in traditional Japanese Kampo medicine and traditional Chinese medicine. Liquorice-containing glycyrrhizin (GL) can cause pseudohyperaldosteronism as a side effect. Previously, we identified 18ß-glycyrrhetyl-3-O-sulfate (3) as a GL metabolite in Eisai hyperbilirubinuria rats (EHBRs) with the dysfunction of multidrug resistance-related protein (Mrp2). We speculated that 3 was associated with the onset of liquorice-induced pseudohyperaldosteronism, because it was mainly detected in serum of patients with suspected to have this condition. However, it is predicted that other metabolites might exist in the urine of EHBRs orally treated with glycyrrhetinic acid (GA). We explored other metabolites in the urine of EHBRs, and investigated the pharmacokinetic profiles of the new metabolite in EHBRs and normal Sprague-Dawley rats. We further analyzed the serum concentrations of the new metabolite in the patients of pseudohyperaldosteronism. Finally, we developed the analyzing method of these metabolites as a preventive biomarker for the onset of pseudohyperaldosteronism using an enzyme-linked immunosorbent assay (ELISA). We isolated a new GL metabolite, 18ß-glycyrrhetyl-3-O-sulfate-30-O-glucuronide (4). Compound 4 significantly inhibited rat type-2 11ß-hydroxysteroid dehydrogenase (11ß-HSD2) and was a substrate of both organic anion transporter (OAT) 1 and OAT3. Compound 4 was also detected in the serum of patients with suspected pseudohyperaldosteronism at an approximately 10-fold lower concentrations than 3, and these concentrations were positively correlated. Compound 4 showed a lower serum concentration and weaker inhibitory titer on 11ß-HSD2 than 3. We developed an enzyme-linked immunosorbent assay system using an anti-18ß-glycyrrhetyl-3-O-glucuronide (3MGA) monoclonal antibody to measure the serum concentration of 3 to facilitate the measurement of biomarkers to predict the onset of pseudohyperaldosteronism. Although we found 4 as the secondary candidate causative agent, 3 could be the main potent preventive biomarker of liquorice-induced pseudohyperaldosteronism. Compound 3 was detected in serum at a higher concentration than GA and 4, implying that 3 may be a pharmacologically active ingredient mediating not only the development of pseudohyperaldosteronism but anti-inflammatory effects in humans administered GL or other liquorice-containing preparations.