BackgroundGreen tea has various health promotion effects. Although there are numerous tea cultivars, little is known about the differences in their nutraceutical properties. Metabolic profiling techniques can provide information on the relationship between the metabolome and factors such as phenotype or quality. Here, we performed metabolomic analyses to explore the relationship between the metabolome and health-promoting attributes (bioactivity) of diverse Japanese green tea cultivars.Methodology/Principal FindingsWe investigated the ability of leaf extracts from 43 Japanese green tea cultivars to inhibit thrombin-induced phosphorylation of myosin regulatory light chain (MRLC) in human umbilical vein endothelial cells (HUVECs). This thrombin-induced phosphorylation is a potential hallmark of vascular endothelial dysfunction. Among the tested cultivars, Cha Chuukanbohon Nou-6 (Nou-6) and Sunrouge (SR) strongly inhibited MRLC phosphorylation. To evaluate the bioactivity of green tea cultivars using a metabolomics approach, the metabolite profiles of all tea extracts were determined by high-performance liquid chromatography-mass spectrometry (LC-MS). Multivariate statistical analyses, principal component analysis (PCA) and orthogonal partial least-squares-discriminant analysis (OPLS-DA), revealed differences among green tea cultivars with respect to their ability to inhibit MRLC phosphorylation. In the SR cultivar, polyphenols were associated with its unique metabolic profile and its bioactivity. In addition, using partial least-squares (PLS) regression analysis, we succeeded in constructing a reliable bioactivity-prediction model to predict the inhibitory effect of tea cultivars based on their metabolome. This model was based on certain identified metabolites that were associated with bioactivity. When added to an extract from the non-bioactive cultivar Yabukita, several metabolites enriched in SR were able to transform the extract into a bioactive extract.Conclusions/SignificanceOur findings suggest that metabolic profiling is a useful approach for nutraceutical evaluation of the health promotion effects of diverse tea cultivars. This may propose a novel strategy for functional food design.

Delphinidin is a member of the anthocyanidin class of plant pigments. We examined the effects of delphinidin on muscle atrophy. Oral administration of delphinidin suppressed the muscle weight loss induced by mechanical unloading. Microarray analysis showed that delphinidin suppresses the upregulation of oxidative stress-related gene expression, including the expression of Cbl-b. Thus, delphinidin may prevent unloading-mediated muscle atrophy.

We show that an extract of a new green tea cultivar Cha Chuukanbohon Nou6 (Nou6), which rich in anthocyanin, has an inhibitory effect on myosin regulatory light chain (MRLC) phosphorylation in rat aortic smooth muscle cell line A7r5. The inhibitory effect of Nou6 extract was abolished when the extract was treated with polyvinylpyrrolidone which traps polyphenols. However, none of the major tea constituents tested had an inhibitory effect. In addition, we found that Nou6 extract had a strong effect on reducing the phosphorylation of myosin phosphatase targeting subunit 1 at Thr696. These results suggest that Nou6 extract inhibits MRLC phosphorylation induced by thrombin through activating myosin light chain phosphatase, and polyphenols in the tea extract may contribute to this effect.Keywords: anthocyanin, smooth muscle cell, myosin regulatory light chain, polyphenol, myosin light chain phosphatase, myosin phosphatase targeting subunit 1 *To whom correspondence should be addressed. E-mail: tatibana@agr.kyushu-u.ac.jp IntroductionContractile state of vascular smooth muscle cells is the primary determinant of blood vessel tone (Ogita et al., 2003). Contractile state has been shown to be directly related to the phosphorylation of myosin regulatory light chain (MRLC) at Ser-19 (Murthy et al., 2006). MRLC phosphorylation controls the activity of myosin II, a major motor protein in animal cells, which is involved in a wide range of processes, including muscle contraction, cell locomotion, cell division, and receptor capping (Maciver, 1996). Phosphorylation of MRLC is regulated by two classes of enzymes: myosin light chain kinase (MLCK) and myosin light chain phosphatase (MLCP) (Somlyo, Somlyo, 2003). MLCP is composed with three subunits: a 37-kDa catalytic subunit, a 20-kDa subunit of unknown function, and a myosin phosphatase targeting subunit 1 (MYPT1). The activity of myosin phosphatase is known to be regulated by phosphorylation of MYPT1, and two major sites, Thr696 and Thr853, have been extensively investigated and identified as inhibitory sites (Hartshorne et al., 2004).Tea (Camellia sinensis L.) has been consumed worldwide for a long time. Among the green tea polyphenols, (-)-epigallocatechin-3-O-gallate (EGCG) is the most abundant and most active polyphenol in inhibiting experimental carcinogenesis and related reactions (Yang et al., 2009). Recently, we have identified 67-kDa laminin receptor (67LR) as a cell surface EGCG receptor that mediates the anticancer action of EGCG (Tachibana et al., 2004). We also reported that EGCG induced reduction of the phosphorylation of MRLC at Thr18/ Ser19 through 67LR in HeLa cells (Umeda et al., 2005), human basophilic KU812 cells (Fujimura et al., 2006), mouse melanoma B16 cells , and human colon adenocarcinoma Caco-2 cells . We found that MYPT1 phosphorylation at Thr696, which inhibits myosin phosphatase and promotes MRLC phosphorylation, was reduced in response to EGCG .New green tea cultivars (Cha Chuukanbohon Nou6 and Sunrouge) are characterized as having a large anthocy...

Background: Patients recovering from COVID-19 often suffer long-term Long-COVID (e.g., depression, poor concentration, anxiety, sleep disturbances, and fatigue). Similar symptoms also rarely seem to occur after COVID-19 vaccination. There is still no effective treatment for these symptoms. We have had a clinical experience that patients presenting with psychiatric/physical symptoms due to COVID-19 or COVID-19 vaccination (defined as Long-COVID and Post-Vaccine patients) often recover after transcranial magnetic stimulation (TMS) and that TMS poorly heals depression in strongly fatigued patients. Aims: 1. Determine whether there are differences in background characteristics and symptoms between Long-COVID and Post-Vaccine patients; 2. Examine whether TMS led to an improvement in their symptoms; 3. Test the involvement of fatigue in the recovery of depression of Long-COVID and Post-Vaccine patients with TMS. Methods: We conducted a retrospective analysis using the medical records of the outpatient clinic of Tokyo TMS Clinic. Results: 1. We found no differences in initial symptoms and courses of treatment between Long-COVID and Post-Vaccine patients. 2. All psychiatric/physical symptom scores after TMS were significantly better than before. Though these results are of before-and-after studies, numerous reports have suggested that TMS effectively improves depression, insomnia, anxiety, and related neuropsychiatric symptoms, which were also primary complaints of patients in this study. We thus attributed the improvement in QIDS, PHQ9 (Both indices of depression), and GAD7 (anxiety indicator) to TMS. 3. The recovery rate of depression in Long-Covid and Post-Vaccine patients with TMS decreased with the severity of fatigue. Conclusions: This is the first report to elucidate the efficacy of TMS and the factors affecting it for psychiatric symptoms after COVID-19 and COVID-19 vaccination. Our study may lead to further validation of the effectiveness and mechanisms of TMS in patients suffering from Long-COVID and COVID-19 vaccine long-term adverse reactions.

COVID-19’s long-term effects, known as Long-COVID, present psychiatric and physical challenges in recovered patients. Similarly, rare long-term post-vaccination side effects, resembling Long-COVID, are emerging (called Post-Vaccine). However, effective treatments for both conditions are scarce. Our clinical experience suggests that transcranial magnetic stimulation (TMS) often aids recovery in Long-COVID and Post-Vaccine patients. However, its effectiveness is reduced in patients with severe fatigue. Therefore, we retrospectively analysed Tokyo TMS Clinic’s outpatient records (60 in total; mean age, 38 years) to compare Long-COVID and post-vaccine patients’ characteristics and symptoms, assess the impact of TMS on their symptoms, and investigate the role of fatigue in depression recovery with TMS. The primary outcome was the regression coefficient of the initial fatigue score on depression score improvement using TMS. Secondary outcomes included psychiatric/physical scores before and after TMS and their improvement rates. We found no differences in the initial symptoms and background factors between Long-COVID and Post-Vaccine patients. After ten TMS sessions, all psychiatric and physical symptom scores improved significantly. TMS improves depression, insomnia, anxiety, and related neuropsychiatric symptoms, which were the primary complaints in this study. Thus, we conclude that TMS improves depression and anxiety. The effectiveness of TMS in treating depression in Long-COVID and Post-Vaccine patients decreased as fatigue severity increased. In conclusion, TMS relieved depressive symptoms following COVID-19 and vaccination; however, fatigue may hinder its effectiveness.