Young-Shick Hong

Metabolomic unveiling of a diverse range of green tea (Camellia sinensis) metabolites dependent on geography

Numerous factors such as geographical origin, cultivar, climate, cultural practices, and manufacturing processes influence the chemical compositions of tea, in the same way as growing conditions and grape variety affect wine quality. However, the relationships between these factors and tea chemical compositions are not well understood. In this study, a new approach for non-targeted or global analysis, i.e., metabolomics, which is highly reproducible and statistically effective in analysing a diverse range of compounds, was used to better understand the metabolome of Camellia sinensis and determine the influence of environmental factors, including geography, climate, and cultural practices, on tea-making. We found a strong correlation between environmental factors and the metabolome of green, white, and oolong teas from China, Japan, and South Korea. In particular, multivariate statistical analysis revealed strong inter-country and inter-city relationships in the levels of theanine and catechin derivatives found in green and white teas. This information might be useful for assessing tea quality or producing distinct tea products across different locations, and highlights simultaneous identification of diverse tea metabolites through an NMR-based metabolomics approach.

Nicotinamide Riboside Ameliorates Hepatic Metaflammation by Modulating NLRP3 Inflammasome in a Rodent Model of Type 2 Diabetes.

Low-grade chronic inflammation (metaflammation) is a major contributing factor for the onset and development of metabolic diseases, such as type 2 diabetes, obesity, and cardiovascular disease. Nicotinamide riboside (NR), which is present in milk and beer, is a functional vitamin B3 having advantageous effects on metabolic regulation. However, the anti-inflammatory capacity of NR is unknown. This study evaluated whether NR modulates hepatic nucleotide binding and oligomerization domain-like receptor family, pyrin domain containing 3 (NLRP3) inflammasome. Male, 8-week-old KK/HlJ mice were allocated to the control or NR group. NR (100 mg/kg/day) or vehicle (phosphate-buffered saline) was administrated by an osmotic pump for 7 days. Glucose control, lipid profiles, NLRP3 inflammasome, and inflammation markers were analyzed, and structural and histological analyses were conducted. NR treatment did not affect body weight gain, food intake, and liver function. Glucose control based on the oral glucose tolerance test and levels of serum insulin and adiponectin was improved by NR treatment. Among tested lipid profiles, NR lowered the total cholesterol concentration in the liver. Histological and structural analysis by hematoxylin and eosin staining and transmission electron microscopy, respectively, showed that NR rescued the disrupted cellular integrity of the mitochondria and nucleus in the livers of obese and diabetic KK mice. In addition, NR treatment significantly improved hepatic proinflammatory markers, including tumor necrosis factor-alpha, interleukin (IL)-6, and IL-1. These ameliorations were accompanied by significant shifts of NLRP3 inflammasome components (NLRP3, ASC, and caspase1). These results demonstrate that NR attenuates hepatic metaflammation by modulating the NLRP3 inflammasome.

Inhibitory Effect of Epigallocatechin Gallate on the Virulence of Clostridium difficile PCR Ribotype 027

Clostridium difficile infection (CDI) is the most prevalent cause of health-care-associated infections. CDI-related health-care costs and deaths are both increasing annually on a global scale. C. difficile have been reported in food products in Canada, Europe, and the United States; however, the systematic transmission of C. difficile between humans and animals is yet to be understood. Because of the limitations of current therapeutic options, there is a need for the development of new patient treatments. Epigallocatechin gallate (EGCG) is a major catechin compound found in green tea extracts and exhibits antioxidant and antimicrobial activities. This study was conducted to investigate the inhibitory effects of EGCG on the expression of virulence genes in C. difficile and in C. difficile-associated diseases by inhibition of quorum sensing. The protein expression of autoinducer-2 (AI-2) was evaluated by AI-2 activity. EGCG at various concentrations had an inhibitory effect on AI-2 production, especially at 10 μg/mL. EGCG also significantly repressed the transcription of virulence genes, including luxS and tcdA, and prolonged the survival of Caenorhabditis elegans infected with C. difficile. Furthermore, treatment with EGCG effectively protected C. difficile-infected mice from C. difficile-induced death. Histological analysis of the colon and cecum of these mice revealed that EGCG protected tissues of the lower intestinal tract from damage. EGCG exerted growth-inhibitory and bactericidal activities on C. difficile in C. difficile-infected mice. Our results suggest that EGCG has significant antipathogenic effects on C. difficile and can be used to prevent or treat C. difficile-associated diseases or C. difficile infections.

Metabolic phenotyping of various tea (Camellia sinensis L.) cultivars and understanding of their intrinsic metabolism

Recently, we selected three tea (Camellia sinensis) cultivars that are rich in taste, epigallocatechin-3-O-gallate (EGCG) and epigallocatechin-3-O-(3-O-methyl)-gallate (EGCG3″Me) and then cultivated them through asexual propagation by cutting in the same region. In the present study, proton nuclear magnetic resonance (1H NMR)-based metabolomics was applied to characterize the metabotype and to understand the metabolic mechanism of these tea cultivars including wild type tea. Of the tea leaf metabolite variations, reverse associations of amino acid metabolism with catechin compound metabolism were found in the rich-taste, and EGCG- and EGCG3″Me-rich tea cultivars. Indeed, the metabolism of individual catechin compounds in the EGCG3″Me-rich cultivar differed from those of other tea cultivars. The current study highlights the distinct metabolism of various tea cultivars newly selected for cultivation and the important role of metabolomics in understanding the metabolic mechanism. Thus, comprehensive metabotyping is a useful method to assess and then develop a new plant cultivar.

Single-walled carbon nanotubes disturbed the immune and metabolic regulation function 13-weeks after a single intratracheal instillation.

Due to their unique physicochemical properties, the potential health effects of single-walled carbon nanotubes (SWCNTs) have attracted continuous attention together with their extensive application. In this study, we aimed to identify local and systemic health effects following pulmonary persistence of SWCNTs. As expected, SWCNTs remained in the lung for 13 weeks after a single intratracheal instillation (50, 100, and 200μg/kg). In the lung, the total number of cells and the percentages of lymphocytes and neutrophils significantly increased at 200μg/kg compared to the control, and the Th1-polarized immune response was induced accompanying enhanced expression of tissue damage-related genes and increased release of chemokines. Additionally, SWCNTs enhanced the expression of antigen presentation-related proteins on the surface of antigen-presenting cells, however, maturation of dendritic cells was inhibited by their persistence. As compared to the control, a significant increase in the percentage of neutrophils and a remarkable decrease of BUN and potassium level were observed in the blood of mice treated with the highest dose. This was accompanied by the down-regulation of the expression of antigen presentation-related proteins on splenocytes. Moreover, protein and glucose metabolism were disturbed with an up-regulation of fatty acid β-oxidation. Taken together, we conclude that SWCNTs may induce adverse health effects by disturbing immune and metabolic regulation functions in the body. Therefore, careful application of SWCNTs is necessary for the enforcement of safety in nano-industries.

Metabolic phenotyping of berries in different six grape (Vitis vinifera) cultivars

Metabolic behaviors of different grapevine cultivars, grown in the same greenhouse, were characterized through a 1H NMR-based metabolomic approach. Pattern recognition, including the principal component analysis, revealed clear dependence of the grape metabolome on the grape cultivar. Interestingly, high accumulations of proline in the purebred grape cultivars of Cabernet Sauvignon, Merlot, and Chardonnay were found, whereas the proline levels were depleted in the crossbred grape cultivars of Steuben, Campbell Early (C. E.), and Seibel. Intrinsic levels of alanine, glutamine, and trans-feruloyl derivative were highest in C. E. cultivar, which grows easily in a wild vineyard, particularly in Korea, suggesting that their levels play important roles in the improvement of resistance or adaptation of the plant to environmental stress, such as freezing stress during the winter season in Korea. The present study highlights that metabolomics is a powerful approach for better understanding the differences of intrinsic metabolic variables of grape berries among various grape cultivars and their associations with the plant physiological mechanisms.

Impact of germination on the structures and in vitro digestibility of starch from waxy brown rice

The in vitro digestibility as well as the molecular and crystalline structures of waxy rice starches isolated from brown rice, germinated brown rice (GBR), ultrasonicated GBR, and heat-moisture treated GBR were investigated. The germinated brown rice starch (GBRS) had a lower average molecular weight and a higher proportion of DP 6-12 in amylopectin than brown rice starch (BRS). The relative crystallinity, intensity ratio of the band at 1,047 cm(-1) and 1,022 cm(-1), gelatinization temperature and pasting temperature of waxy rice starch were reduced by germination. However, the ultrasonication and heat-moisture treatment of GBRS increased the relative crystallinity and gelatinization temperature. The digestibility of starch from brown waxy rice was increased by germination. The rapidly digestible starch (RDS), slowly digestible starch (SDS) and resistant starch (RS) contents were 50.5%, 42.4%, and 7.1% in BRS, and 69.0%, 27.9% and 3.1% in GBRS, respectively. The ultrasonication and heat-moisture treatment of GBRS reduced RDS content and increased RS content in raw and gelatinized starches. The decrease in starch digestibility of cooked GBR was more pronounced after heat-moisture treatment than after ultrasonication.

Diverse Metabolite Variations in Tea (Camellia sinensis L.) Leaves Grown Under Various Shade Conditions Revisited: A Metabolomics Study

With the increase of tea (Camellia sinensis) consumption, its chemical or metabolite compositions play a crucial role in the determination of tea quality. In general, metabolite compositions of fresh tea leaves including shoots depend on plucking seasons and tea cultivators. Therefore, choosing a specific plucking time of tea leaves can provide use-specified tea products. Artificial control of tea growing, typically shade treatments, can lead to significant changes of the tea metabolite compositions. However, metabolic characteristics of tea grown under various shade treatment conditions remain unclear. Therefore, the objective of the current study was to explore effects of various shade conditions on metabolite compositions of tea through a 1H NMR-based metabolomics approach. It was noteworthy that the levels of catechins and their derivatives were only influenced at the initial time of shade treatments while most amino acids were upregulated as amounts of shade and periods were increased: that is, the levels of alanine, asparagine, aspartate, isoleucine, threonine, leucine, and valine in fresh tea leaves were conspicuously elevated when shade levels were raised from 90% to 100% and when period of shade treatments was increased by 20 days. Such increased synthesis of amino acids along with large reductions of glucose level reflected carbon starvation under the dark conditions, indicating remarkable proteolysis in the chloroplast of tea leaves. This study provides important information about making amino acid-enhanced tea products based on global characteristics of diverse tea leaf metabolites induced by various shade treatment conditions.

Metabolomics approach for understanding geographical dependence of soybean leaf metabolome

The soybean plant (Glycine max) is widely used as an ingredient in various foods, nutraceuticals and cosmetics, due to their diverse bioactive compounds. Their metabolic compositions are likely affected by environmental conditions during growth. To investigate the influence of different environmental conditions on the metabolite composition of soybean leaves, we cultivated soybean (G. max Sinhwa) in the southernmost island and volcanic region of Korea, and in the central section and limestone region of the Korean peninsula. Comprehensive metabolite variations of their leaves were analyzed through 1H NMR-based metabolomics approach. With marked differences in soil compositions and climatic conditions between the two growing areas, differences in accumulations of pinitol and diverse flavonoids were noted between the soybean leaves, reflecting the distinct metabolism of soybean plants for physiological adaptation toward different environmental conditions. Therefore, the current study highlights the geographical dependences of diverse soybean leaf metabolites for developing biofunction-enhanced soybean products.

Metabotyping of rice (Oryza sativa L.) for understanding its intrinsic physiology and potential eating quality

Rice (Oryza sativa L.), the major staple food in many countries, has genetic diversity adapted to different environmental conditions. However, metabolic traits about diverse rice plants are rarely discovered. In the present study, rice leaves and grains were collected at whole growth stages from late (LMC) and early (EMC) maturing cultivars. Metabolic dependences of rice plants on both growth and cultivar were investigated in their leaves and grains through NMR-based metabolomics approach. Rice leaf metabolome were differently regulated between two rice cultivars, thereby affecting variations of rice grain metabolome. Sucrose levels in leaves of EMC were markedly decreased compared to those in LMC, and more accumulations of sucrose, amino acids and free fatty acids were found in grains of EMC. These distinct metabolisms between EMC and LMC rice cultivars were associated with temperature during their growing seasons and might affect the eating quality of rice. The current study highlights that metabolomic approach of rice leaves and grains could lead to better understanding of the relationship between their distinct metabolisms and environmental conditions, and provide novel insights to metabolic qualities of rice grains.