Cesium uptake and translocation from tea cutting roots (Camellia sinensis L.).

To estimate the uptake of radiocesium (137Cs) by tea plant roots, 1-year-old rooted tea cuttings (Camellia sinensis L. cv. Yabukita) at the time of bud opening were cultivated hydroponically for 27 days in pots containing nutrient solutions with or without 137CsCl (600 Bq mL-1). Total 137Cs radioactivity of whole tea plants were 6.1 kBq g-1 dry weight. The plant/solution 137Cs transfer factors of different tissues were in the range of 2.6 (in mature leaves) to 28.2 mL g-1 dry weight (in roots), which were lower than those reported in wheat and spinach. In total, 69% of 137Cs remained in roots and 31% was transported from roots to shoots. The results indicated that 137Cs was preferentially translocated to new shoots, which are used for manufacturing tea, over mature leaves.

Potential of spectroscopic analyses for non-destructive estimation of tea quality-related metabolites in fresh new leaves.

Spectroscopic sensing provides physical and chemical information in a non-destructive and rapid manner. To develop non-destructive estimation methods of tea quality-related metabolites in fresh leaves, we estimated the contents of free amino acids, catechins, and caffeine in fresh tea leaves using visible to short-wave infrared hyperspectral reflectance data and machine learning algorithms. We acquired these data from approximately 200 new leaves with various status and then constructed the regression model in the combination of six spectral patterns with pre-processing and five algorithms. In most phenotypes, the combination of de-trending pre-processing and Cubist algorithms was robustly selected as the best combination in each round over 100 repetitions that were evaluated based on the ratio of performance to deviation (RPD) values. The mean RPD values were ranged from 1.1 to 2.7 and most of them were above the acceptable or accurate threshold (RPD = 1.4 or 2.0, respectively). Data-based sensitivity analysis identified the important hyperspectral regions around 1500 and 2000 nm. Present spectroscopic approaches indicate that most tea quality-related metabolites can be estimated non-destructively, and pre-processing techniques help to improve its accuracy.

Dissection of hyperspectral reflectance to estimate nitrogen and chlorophyll contents in tea leaves based on machine learning algorithms.

Nondestructive techniques for estimating nitrogen (N) status are essential tools for optimizing N fertilization input and reducing the environmental impact of agricultural N management, especially in green tea cultivation, which is notably problematic. Previously, hyperspectral indices for chlorophyll (Chl) estimation, namely a green peak and red edge in the visible region, have been identified and used for N estimation because leaf N content closely related to Chl content in green leaves. Herein, datasets of N and Chl contents, and visible and near-infrared hyperspectral reflectance, derived from green leaves under various N nutrient conditions and albino yellow leaves were obtained. A regression model was then constructed using several machine learning algorithms and preprocessing techniques. Machine learning algorithms achieved high-performance models for N and Chl content, ensuring an accuracy threshold of 1.4 or 2.0 based on the ratio of performance to deviation values. Data-based sensitivity analysis through integration of the green and yellow leaves datasets identified clear differences in reflectance to estimate N and Chl contents, especially at 1325-1575 nm, suggesting an N content-specific region. These findings will enable the nondestructive estimation of leaf N content in tea plants and contribute advanced indices for nondestructive tracking of N status in crops.

Genomic predictions and genome-wide association studies based on RAD-seq of quality-related metabolites for the genomics-assisted breeding of tea plants.

Effectively using genomic information greatly accelerates conventional breeding and applying it to long-lived crops promotes the conversion to genomic breeding. Because tea plants are bred using conventional methods, we evaluated the potential of genomic predictions (GPs) and genome-wide association studies (GWASs) for the genetic breeding of tea quality-related metabolites using genome-wide single nucleotide polymorphisms (SNPs) detected from restriction site-associated DNA sequencing of 150 tea accessions. The present GP, based on genome-wide SNPs, and six models produced moderate prediction accuracy values (r) for the levels of most catechins, represented by ( -)-epigallocatechin gallate (r = 0.32-0.41) and caffeine (r = 0.44-0.51), but low r values for free amino acids and chlorophylls. Integrated analysis of GWAS and GP detected potential candidate genes for each metabolite using 80-160 top-ranked SNPs that resulted in the maximum cumulative prediction value. Applying GPs and GWASs to tea accession traits will contribute to genomics-assisted tea breeding.

Analyses of single nucleotide polymorphisms identified by ddRAD-seq reveal genetic structure of tea germplasm and Japanese landraces for tea breeding.

To obtain genetic information about the germplasm of tea (Camellia sinensis L.) in Japan, 167 accessions including 138 var. sinensis (96 Japanese var. sinensis and 42 exotic var. sinensis) and 29 Assam hybrids were analyzed using single nucleotide polymorphisms (SNPs) markers identified by double-digest restriction-site-associated DNA sequencing (ddRAD-seq) analysis. Approximately 10,000 SNPs were identified by ddRAD-seq and were mapped across the whole genome. The 167 tea accessions were classified into three genetic subgroups: (1) Japanese var. sinensis; (2) Japanese and exotic var. sinensis; (3) Assam hybrids and exotic var. sinensis. Leaf morphology varied widely within each genetic subgroups. The 96 Japanese var. sinensis were classified into four genetic subgroups as follows; two subgroups of Shizuoka (the largest tea production region) landraces, Uji (most ancient tea production region) landraces, and the pedigree of 'Yabukita', the leading green tea cultivar in Japan. These results indicated that the SNP markers obtained from ddRAD-seq are a useful tool to investigate the geographical background and breeding history of Japanese tea. This genetic information revealed the ancestral admixture situation of the 'Yabukita' pedigree, and showed that the genome structure of 'Yabukita' is clearly different from those of other Japanese accessions.

Stress-Reducing Function of Matcha Green Tea in Animal Experiments and Clinical Trials.

Theanine, a major amino acid in green tea, exhibits a stress-reducing effect in mice and humans. Matcha, which is essentially theanine-rich powdered green tea, is abundant in caffeine. Caffeine has a strong antagonistic effect against theanine. The stress-reducing effect of matcha was examined with an animal experiment and a clinical trial. The stress-reducing effect of matcha marketed in Japan and abroad was assessed based on its composition. The stress-reducing effect of matcha in mice was evaluated as suppressed adrenal hypertrophy using territorially-based loaded stress. High contents of theanine and arginine in matcha exhibited a high stress-reducing effect. However, an effective stress-reducing outcome was only possible when the molar ratio of caffeine and epigallocatechin gallate (EGCG) to theanine and arginine was less than two. Participants (n = 39) consumed test-matcha, which was expected to have a stress-reducing effect, or placebo-matcha, where no effect was expected. Anxiety, a reaction to stress, was significantly lower in the test-matcha group than in the placebo group. To predict mental function of each matcha, both the quantity of theanine and the ratios of caffeine, EGCG, and arginine against theanine need to be verified.

Deep Decompression of the Lateral Femoral Cutaneous Nerve Under Local Anesthesia.

OBJECTIVE: Meralgia paresthetica is a mononeuropathy of the lateral femoral cutaneous nerve (LFCN) caused by compression around the inguinal ligament. We report a surgical alternative for the treatment of meralgia paresthetica under local anesthesia and its outcomes. METHODS: We operated on 12 patients with unilateral meralgia paresthetica whose age at surgery ranged from 62 to 75 years. The mean postoperative follow-up period was 19 months. Microsurgical deep decompression of LFCN was performed with the patient under local anesthesia. Clinical outcomes of surgical treatment were assessed based on the patient's most recent follow-up visit and were classified into 3 categories: complete, partial, or no relief of symptoms. Symptoms of pain or numbness in the anterolateral part of the thigh were evaluated, using a visual analog scale, before surgery and after surgery, i.e., at the most recent follow-up visit. RESULTS: All patients reported symptom improvement: complete relief in 9 patients (75%) and partial relief in 3 patients (25%). In the 3 patients with partial relief, the remaining symptoms did not affect their daily living. Overall, the visual analog scale scores were significantly improved in all patients (P < 0.05), and no patient experienced postoperative recurrence of their symptoms at the time of the last follow-up visit. CONCLUSIONS: Symptoms of meralgia paresthetica can resemble those of a lumbosacral disorder. Microsurgical deep decompression under local anesthesia produces good surgical outcomes. The use of local anesthesia contributes not only to reduction of pain during surgery but also eliminates excessive surgical procedures and reduces the duration of hospital stay.

Quantitative validation of nicotine production in tea (Camellia sinensis L.).

Endogenous nicotine was confirmed to be present in tea plants (Camellia sinensis L.) by liquid chromatography-tandem mass spectrometry of tea samples from tea-producing regions in six Asian countries. All samples contained nicotine (0.011-0.694 µg g-1 dry weight). Nicotine contents remained constant during manufacturing of green, oolong and black teas, implying that nicotine is stable against heating, drying, enzymatic oxidation and mechanical damage during processing. Flower buds and seeds of cultivar Yabukita also contained nicotine (0.030-0.041 µg g-1 dry weight). A comparison of two cultivars revealed that higher nicotine contents were found in the black tea cultivar Benifuki. All plant parts of hydroponic Yabukita contained nicotine (0.003-0.013 µg g-1 dry weight). Tea cells cultured in B5 medium as well as roots and stems of tea seedlings contained nicotine levels similar to those of new leaves from field-grown plants. Although the levels of endogenous nicotine in tea plants are extremely low and sample contamination cannot be discounted, these levels exceed the maximum acceptable limit in Japan (0.01 µg g-1 dry weight).

[Chronic Low Back Pain Associated with Gluteus Medius Muscle:A Case Report].

The clinical features and etiology of low back pain(LBP)and buttock pain(BuP)has been poorly understood. We report a case of long-term BuP that was successfully treated with gluteus medius muscle(GMeM)decompression under local anesthesia. A 71-year-old man was referred to our hospital because of long-term BuP and claudication. Left BuP that radiated to the left thigh was observed. The pain was mostly triggered by palpation at the middle of the iliac crest and greater trochanter. Lumbar and pelvic radiograms showed no significant lesions. Lumbar magnetic resonance imaging revealed a mild lumbar spinal canal stenosis at the L4/L5 segment. Based on the evidence of a trigger point and pain relieved after GMeM block injection, we made a diagnosis of GMeM pain. Although several GMeM block injections relieved his pain, the analgesic effect was transient and the claudication remained. Then, we decided to perform GMeM decompression. We made a 5-cm-long skin incision across the trigger point on the buttock. After confirming a wide exposure of the gluteal aponeurosis over the GMeM, we cut and opened it for sufficient GMeM decompression, and the GMeM expansion was confirmed. After surgery, his symptoms immediately improved. No evidence of recurrence was observed 6 months after his treatment. For the treatment of LBP and BuP, GMeM pain would be considered a causative factor. We report that it can be treated with a less invasive surgical technique, which would contribute to good clinical outcome.

Radiocesium uptake through leaf surfaces of tea plants (Camellia sinensis L.).

To clarify the source of radiocesium detected in newly emerged tea leaves contaminated just before the time of bud opening by fallout of radionuclides from Fukushima Dai-ichi Nuclear Power Plant, 137CsCl solution (0.185 M Bq mL-1) was applied to the front or the backside surfaces of mature leaves of tea plant (Camellia sinensis L. cv. Yabukita) at the time of bud opening. A 21 days after foliar application, the buds had grown and developed to the three- or four-leaf stage. In the front treatment, almost all (95%) of the applied 137Cs was present in the mature leaves (hot mother leaves). In the backside treatment, 68% of applied 137Cs also remained in hot mother leaves, but 22% and 10% was found in the new shoots attached to hot mother leaves and the other parts (non-applied mature leaves, stems and roots), respectively. The images of a hot leaf and its attached new shoots by imaging plate analysis revealed that the results coincided with those of the 137Cs distribution above. These suggested that radiocesium was primarily absorbed from the backside surface of tea leaves through the stoma, and then the greater part was transported to newly emerged tea organs during the new shoot growth period.

Acute Poststroke Depression Is Associated with Thalamic Lesions and Clinical Outcomes: A Case-Control Study.

BACKGROUND: We investigated the role of acute-phase stroke lesions and patient characteristics in poststroke depression (PSD) and its effect on the clinical outcome. PATIENTS AND METHODS: Five and 30 days after admission, 175 patients self-reported their depressive symptoms on the Patient Health Questionnaire-9. We compared the clinical characteristics and outcomes in patients with (n = 41) and without PSD (n = 134). Stroke severity was assessed with the National Institutes of Health Stroke Scale (NIHSS); the modified Rankin Scale (mRS) was used to determine the functional outcome. RESULTS: There was no significant difference between patients with and without PSD in the age, gender ratio, lesion side, and the history of hypertension, diabetes mellitus, alcohol and tobacco use, and previous stroke. Thalamic lesions were significantly associated with PSD (P = .03), although there was no significant difference in both the NIHSS score and the final mRS score of patients with thalamic lesions. Backward stepwise logistic regression analysis showed that a higher NIHSS score and thalamic lesions were independent predictors of PSD. Total hospitalization was significantly longer in patients with PSD. At the time of admission, the NIHSS score was significantly higher in patients who developed moderate to severe PSD than in those with mild PSD or without PSD. CONCLUSIONS: PSD in the acute phase was associated with thalamic lesions and severe stroke. Hospitalization was significantly longer in patients with PSD and their functional disability was more severe, suggesting that PSD played a role in the unsatisfactory results of poststroke rehabilitation.

Reduced Stress and Improved Sleep Quality Caused by Green Tea Are Associated with a Reduced Caffeine Content.

Caffeine, one of the main components in green tea, can interfere with sleep and block the effect of theanine. Since theanine, the main amino acid in tea leaves, has significant anti-stress effects in animals and humans, we examined the effects of green tea with lowered caffeine content, i.e., low-caffeine green tea (LCGT), on stress and quality of sleep of middle-aged individuals (n = 20, mean age 51.3 ± 6.7 years) in a double-blind crossover design. Standard green tea (SGT) was used as the control. These teas (≥300 mL/day), which were eluted with room temperature water, were consumed over a period of seven days after a single washout term. The level of salivary α-amylase activity (sAA), a stress marker, was significantly lower in participants that consumed LCGT (64.7 U/mL) than in those that consumed SGT (73.9 U/mL). Sleep quality was higher in participants that consumed a larger quantity of LCGT. In addition, a self-diagnostic check for accumulated fatigue was significantly lower in those participants that consumed LCGT than SGT. These results indicate that LCGT intake can reduce stress in middle-aged individuals and improve their quality of sleep. The reduction in caffeine is suggested to be a valid reason for enhancing the anti-stress effect of green tea.

Anti-stress Effect of Green Tea with Lowered Caffeine on Humans: A Pilot Study.

Theanine, an amino acid in tea, has significant anti-stress effects on animals and humans. However, the effect of theanine was blocked by caffeine and gallate-type catechins, which are the main components in tea. We examined the anti-stress effect of green tea with lowered caffeine, low-caffeine green tea, on humans. The study design was a single-blind group comparison and participants (n=20) were randomly assigned to low-caffeine or placebo tea groups. These teas (≥500 mL/d), which were eluted with room temperature water, were taken from 1 week prior to pharmacy practice and continued for 10 d in the practice period. The participants ingested theanine (ca. 15 mg/d) in low-caffeine green tea. To assess the anxiety of participants, the state-trait anxiety inventory test was used before pharmacy practice. The subjective stress of students was significantly lower in the low-caffeine-group than in the placebo-group during pharmacy practice. The level of salivary α-amylase activity, a stress marker, increased significantly after daily pharmacy practice in the placebo-group but not in the low-caffeine-group. These results suggested that the ingestion of low-caffeine green tea suppressed the excessive stress response of students. This study was registered at the University Hospital Medical Information Network (ID No. UMIN14942).

Anti-stress effects of drinking green tea with lowered caffeine and enriched theanine, epigallocatechin and arginine on psychosocial stress induced adrenal hypertrophy in mice.

BACKGROUND: Theanine, an amino acid in tea, has significant anti-stress effects on animals and humans. However, the anti-stress effects of drinking green tea have not yet been elucidated. HYPOTHESIS/PURPOSE: The present study aimed to explore anti-stress effects of green tea and roles of tea components in a mouse model of psychosocial stress. STUDY DESIGN: We examined anti-stress effects of three types of green teas, theanine-rich "Gyokuro", standard "Sencha", and Sencha with lowered caffeine (low-caffeine green tea). Furthermore, the roles of tea components such as caffeine, catechins, and other amino acids in anti-stress effects were examined. METHODS: To prepare low-caffeine green tea, plucked new tea leaves were treated with a hot-water spray. Mice were psychosocially stressed from a conflict among male mice under confrontational housing. Mice consumed each tea that was eluted with room temperature water ad libitum. As a marker for the stress response, adrenal hypertrophy was compared with mice that ingested water. RESULTS: Caffeine was significantly lowered by spraying hot-water on tea leaves. While epigallocatechin gallate (EGCG) is the main catechin in tea leaves, epigallocatechin (EGC) was mainly infused into water at room temperature. Adrenal hypertrophy was significantly suppressed in mice that ingested theanine-rich and low-caffeine green tea that were eluted with water at room temperature. Caffeine and EGCG suppressed the anti-stress effects of theanine while EGC and arginine (Arg) retained these effects. CONCLUSION: These results suggest that drinking green tea exhibits anti-stress effects, where theanine, EGC and Arg cooperatively abolish the counter-effect of caffeine and EGCG on psychosocial stress induced adrenal hypertrophy in mice.

Hyperthermotherapy enhances antitumor effect of 5-aminolevulinic acid-mediated sonodynamic therapy with activation of caspase-dependent apoptotic pathway in human glioma.

Sonodynamic therapy (SDT) has shown great potential as an approach for cancer treatment, and hyperthermotherapy (HT) is also a promising cancer therapy. Here, we investigate whether HT could improve the efficacy of SDT and to make a preliminary exploration on potential mechanism. Xenograft tumor was established in nude mice model, and SNB19 and U87MG glioma cell lines were utilized for in vitro experiment. Alamar blue assay was performed to assess cell viability. Optical microscope was used to characterize the morphology changes of the glioma cells induced by SDT and HT treatments. Apoptotic rate, mitochondrial membrane potential (MMP), and intracellular production of reactive oxygen species (ROS) were examined by flow cytometer. The cell apoptosis of tumor tissues were detected by TUNEL assay. Furthermore, the expression of apoptosis-related proteins was detected with Western blot in vitro and immunohistochemistry in vivo. SDT plus HT group could significantly reduce the cell viability with circular-cell morphological change, compared with SDT group, and cell viability was decreased depending on raise of 5-ALA concentration, ultrasound exposure time, and temperature. The results also indicate that HT increased a conspicuous apoptosis, ROS production, and a remarkable loss in MMP induced by 5-ALA-SDT in vitro. Meanwhile, our data also demonstrated that the combined treatment could significantly induce apoptosis and delay tumor growth in vivo. Furthermore, in both in vitro and in vivo experiments, SDT plus HT group expressed significantly higher protein levels of Bax and cleaved caspase-3, 8, and 9 compared to SDT, HT, and control groups and significantly lower protein level of bcl-2 than the other three groups, while the expression of these proteins was unchanged between HT and control groups. HT may provide an important promotion on 5-ALA-SDT and further propose that SDT in combination with HT is a new potential application for the treatment of human glioma.

Neurovascular bundle decompression without excessive dissection for tarsal tunnel syndrome.

Tarsal tunnel syndrome (TTS) is an entrapment neuropathy of the posterior tibial nerve and its branches in the tarsal tunnel. We present our less invasive surgical treatment of TTS in 69 patients (116 feet) and their clinical outcomes. The mean follow-up period was 64.6 months. With the patient under local anesthesia we use a microscope to perform sharp dissection of the flexor retinaculum and remove the connective tissues surrounding the posterior tibial nerve and vessels. To prevent postoperative adhesion and delayed neuropathy, decompression is performed to achieve symptom improvement without excessive dissection. Decompression is considered complete when the patient reports intraoperative symptom abatement and arterial pulsation is sufficient. The sensation of numbness and/or pain and of foreign substance adhesion was reduced in 92% and 95% of our patients, respectively. In self-assessments, 47 patients (68%) reported the treatment outcome as satisfactory, 15 (22%) as acceptable, and 7 (10%) were dissatisfied. Of 116 feet, 4 (3%) required re-operation, initial decompression was insufficient in 2 feet and further decompression was performed; in the other 2 feet improvement was achieved by decompression of the distal tarsal tunnel. Our surgical method involves neurovascular bundle decompression to obtain sufficient arterial pulsation. As we use local anesthesia, we can confirm symptom improvement intraoperatively, thereby avoiding unnecessary excessive dissection. Our method is simple, safe, and without detailed nerve dissection and it prevents postoperative adhesion.

Influence of exogenously applied abscisic acid on carotenoid content and water uptake in flowers of the tea plant (Camellia sinensis).

BACKGROUND: Carotenoids are a major class of plant pigments and fulfill many functions in different organisms that either produce or consume them. Although the color of the stamina of tea (Camellia sinensis) flowers is clearly due to the presence of carotenoids, the carotenoid profile and content remain to be discovered. RESULTS: We investigated the carotenoid profile of tea flowers and determined changes in concentrations over the floral development. The flowers contained oxygenated xanthophylls such as neoxanthin, lutein and zeaxanthin, as well as the hydrocarbons ß-carotene and α-carotene. Flowers of the tea plant contain to vegetables comparable amounts of carotenoids. The content of 9'-cis-epoxycarotenoids, which serve as abscisic acid precursors, as well as changes in concentration of abscisic acid were studied. The concentrations of carotenoids decreased whereas the abscisic acid content increased over the floral development. Exogenously applied S-abscisic acid affected water uptake, flower opening and carotenoid accumulation. CONCLUSION: In summary, this paper reports, for the first time, the carotenoid profile and content of tea flowers. The study revealed that carotenoids in tea flowers are an interesting target in respect of possible applications of tea flower extracts as well as biological functions of abscisic acid during floral development.

Sub-critical water extraction of residual green tea to produce a roasted green tea-like extract.

In this study, we investigated the effects of various temperatures between 140 to 260 degrees C during sub-critical water extraction of residual green tea in making a roasted green tea-like extract, a popular beverage in Japan. Each residual green tea extract was analyzed for sensory properties and antioxidative activities, and this revealed 140 degrees C to be the best extraction temperature. 5-Hydroxymethyl-2-furaldehyde, (+)-catechin, and (-)-epicatechin were found to be the major antioxidative compounds in the 140 degrees C extract, along with theanine and some important amino acids.

Mechanism for the detoxification of aluminum in roots of tea plant (Camellia sinensis (L.) Kuntze).

To determine the mechanism of aluminum (Al) detoxification in the roots of tea plants (Camellia sinensis (L.) Kuntze), the amounts of Al and Al-chelating compounds (fluoride (F), organic acids and catechins) were measured and the chemical forms of Al in root cell extracts were identified by the application of 27Al-nuclear magnetic resonance (NMR) spectroscopy. Tea plants were cultivated in nutrient solutions containing 0, 4, 1.0 and 4.0 mM of Al at pH 4.2 for approximately 10 weeks. The levels of soluble Al, water-soluble oxalate and citrate, but not F, malate or catechins in young roots increased with an increase in the concentration of Al in the treatment solution. The 27Al NMR spectra of root tips and cell sap extracted from root tips that had been treated with Al were almost identical and had four signals, with two (11 and 16 ppm) apparently corresponding to the known chemical shifts of Al-oxalate complexes. In the spectra of cell sap, the resonances at 11 and 16 ppm increased with an increase in the Al contents. These results suggest that the levels of Al-oxalate complexes increased in response to an increase in the Al level, implying that oxalate is a key Al-chelating compound in the mechanism of Al detoxification in the tea root.