Anti-Helicobacter pylori Activity of Six Major Compounds Isolated from Rumex acetosa.

Gastric problems are often caused by the well-known Helicobacter pylori (H. pylori) bacterium. One of the biggest obstacles to the treatment of H. pylori infections is increasing the antibiotic resistance. During our search for naturally derived anti-H. pylori compounds, six major compounds were isolated from the methylene chloride (CH2Cl2) and ethyl acetate (EtOAc) fractions of Rumex acetosa that showed anti-H. pylori activity. Three anthraquinones and three anthraquinone glucosides were identified as the major chemical constituents of the CH2Cl2 and EtOAc fractions, respectively. The chemical structures were identified to be emodin (1), chrysophanol (2), physcion (3), emodin-8-O-ß-d-glucoside (4), chrysophanol-8-O-ß-d-glucoside (5), and physcion-8-O-ß-d-glucoside (6) by UV, 1H NMR, 13C NMR, and mass spectrometry. Anti-H. pylori activity, including the minimum inhibitory concentration (MIC) value of each compound, was evaluated against two H. pylori strains. All isolates exhibited anti-H. pylori activity with different potencies, with an MIC value ranging between 3.13 and 25 µM. However, some variations were found between the two strains. While compound 5 displayed the most potent antibacterial activity with an MIC50 value of 8.60 µM and an MIC90 value of 15.7 µM against H. pylori strain 51, compound 1 exhibited the most potent inhibitory activity against H. pylori strain 43504. The two compounds also showed moderate urease inhibitory activity, with compound 1 demonstrating activity higher than that of compound 5. Furthermore, a molecular docking study revealed the high binding ability of compounds 1 and 5 to the active site of H. pylori urease. The present study suggests that the six anthraquinones isolated from R. acetosa with the whole parts of this plant may be natural candidates for the treatment of H. pylori infection. Further studies are required to determine the exact mechanism of action and to evaluate safety issues in the human body.

Reproducibility of MRI-derived liver surface nodularity score: analysis of patients with repeated MRI in various scanners.

PURPOSE: To assess trans-regional differences, reproducibility across different MRI scanners, and interobserver agreement of liver surface nodularity (LSN) score from routine liver MRI and to evaluate the correlation between LSN score and liver stiffness (LS) value on MR elastography. MATERIALS AND METHODS: Ninety patients who underwent gadoxetic acid-enhanced liver MRI twice using different MRI scanners within a year were evaluated. On axial hepatobiliary phase images, right anterior (LSNRT_ANT), right posterior (LSNRT_POST), and left anterior hepatic surface (LSNLT) were chosen for the quantification of LSN score. Repeated-measures ANOVA, paired t test, Pearson's correlation coefficient analysis, and intraclass correlation coefficient (ICC) were used for statistical analysis. RESULTS: LSN scores from high to low were LSNRT_POST, LSNRT_ANT, and LSNLT, representing trans-regional differences (p < 0.001). Reproducibility of LSN measurement across different MRI scanners was high to excellent (ICC = 0.838-0.921). The mean difference between first and second examinations in LSNRT_ANT, LSNRT_POST, and LSNLT were 0.032 (p = 0.013), 0.002 (p = 0.910), and 0.010 (p = 0.285) for reader 1 and 0.051 (p = 0.004), 0.061 (p = 0.002), and 0.023 (p = 0.005) for reader 2. The first and second examinations were highly correlated in all hepatic regions (r = 0.712-0.839, p < 0.001). There was a low to moderate correlation between LSN score and LS value (r = 0.364-0.592, p ≤ 0.001), which was higher in the chronic hepatitis B (CHB) group than in the non-CHB group in all hepatic regions. CONCLUSIONS: In our study, LSN measurement on liver MRI showed trans-regional differences and excellent reproducibility across different MRI scanners. To use LSN score more widely, standardization of quantification software and selected hepatic regions is needed.

Long-term results of Bernese periacetabular osteotomy using a dual approach in hip dysplasia.

We report the long-term results of Bernese periacetabular osteotomy using a dual approach in hip dysplasia. Fifty-three hips (49 patients, mean age 39.9 years: 13-62 years; bilateral hips: four patients) that underwent periacetabular osteotomy using a dual approach (combined Smith-Peterson and Kocher-Langenbeck techniques) between May 1997 and December 2005 were analyzed in this study. The clinical and radiologic outcomes and complications were analyzed and the final survival rates of the operated hips were investigated with survival analysis curves. Forty-nine hips survived until the final follow-up without arthroplasty, and four hips underwent arthroplasty. The average follow-up period was 11.5 years (8-16 years). The pain visual analogue scale improved from 6.3 to 1.1, while the Harris hip score improved from 61.9 to 91.1. Radiologic findings showed that all cases showed improvements in the center edge angle, acetabular angle, acetabular depth, and femoral head coverage. Two patients underwent intraarticular osteotomy due to a complication, and one patient underwent additional osteotomy due to an under-correction. Three cases showed an asymptomatic nonunion of the superior pubic ramus osteotomy site. One patient developed an avulsion fracture of the anterior superior iliac spine, and none of the cases had an infection or permanent neurologic damage. Kaplan-Meier analysis revealed that the 10-year survival rate was 93% (95% confidence interval [CI] 81-98%) with arthroplasty as the endpoint and 86% (95% CI 70-91%) with the progression of osteoarthritis based on Tönnis osteoarthritis rating as the endpoint. Based on the outcomes of a long-term follow-up of more than 10 years on average, Bernese periacetabular osteotomy via a dual approach was found to be a satisfactory method for lowering the incidence of complications while preserving hips.

Catechin-7-O-α-L-rhamnopyranoside can reduce α-MSH-induced melanogenesis in B16F10 melanoma cells through competitive inhibition of tyrosinase.

Although melanogenesis is a defense mechanism against ultraviolet (UV)-induced skin damage, abnormally excessive melanin production causes pigmentation disorders. Tyrosinase, as a key factor for melanin synthesis, plays an important role in inducing skin pigmentation. Therefore, the inhibition of tyrosinase is crucial in preventing skin pigmentation in the cosmetics and medicine fields. However, the majority of well-known tyrosinase inhibitors have been discontinued due to toxic effects on the skin or lack of selectivity and/or stability. In this study, we evaluated possible anti-melanogenic effects of catechin-7-O-α-L-rhamnopyranoside (C7R) isolated from the stem bark of Ulmus parvifolia, to discover a new tyrosinase inhibitor that has both safety and stability. When C7R was pretreated in B16F10 melanoma cells stimulated by α-melanocyte-stimulating hormone, this compound reduced melanin accumulation and murine tyrosinase activity. In line with these results, C7R inhibits tyrosinase purified from a mushroom in vitro like kojic acid and arbutin. Furthermore, C7R exhibited a competitive inhibition on a Lineweaver-Burk plot. Next, the underlying mechanisms of the C7R-mediated tyrosinase inhibitory effect were sought through docking simulation and pharmacophore analysis between tyrosinase residues and C7R. The results of these analyses showed that C7R had binding energy of -14.5kcal/mol, and indicated that C7R interacts with tyrosinase through an aromatic ring and various hydrophobic and hydrogen bonds. Together, our results suggest that C7R can be applied as a novel natural anti-melanogenic agent that inhibits tyrosinase.

Protective Effect of Carotenoid Extract from Orange-Fleshed Sweet Potato on Gastric Ulcer in Mice by Inhibition of NO, IL-6 and PGE2 Production.

Ipomoea batatas (L.) Lam., Convolvulaceae is widely distributed in Asian areas from tropical to warm-temperature regions. Their tubers are known for their antioxidant, anti-bacterial, anti-diabetic, wound healing, anti-inflammatory, and anti-ulcer activities. The preventive and therapeutic effects of orange-fleshed sweet potato on gastric ulcers have not been investigated. In this study, the carotenoid extract (CE) of orange-fleshed sweet potato was found to protect against gastric ulcers induced by HCl/ethanol in mice. The anti-inflammatory and antioxidant activities of the carotenoid pigment extract were also evaluated as possible evidence of their protective effects. Administration of CE reduced gastric ulcers. Oral administration of CE (100 mg/kg) protected against gastric ulcers by 78.1%, similar to the positive control, sucralfate (77.5%). CE showed potent reducing power and decreased nitric oxide production in a mouse macrophage cell line, RAW 264.7, in a concentration-dependent manner. The production of the inflammatory cytokine interleukin-6 and prostaglandin E2 was also reduced by CE in a dose-dependent manner. The high carotenoid content of orange-fleshed sweet potato could play a role in its protective effect against gastric ulcers. This result suggests the possibility of developing functional products using this nutrient-fortified material.

Anatomical and Chemical Characterization of Ulmus Species from South Korea.

Ulmus species (Ulmaceae) are large deciduous trees distributed throughout Korea. Although their root and stem bark have been used to treat gastrointestinal diseases and wounds in folk medicine, commercial products are consumed without any standardization. Therefore, we examined anatomical and chemical differences among five Ulmus species in South Korea. Transverse sections of leaf, stem, and root barks were examined under a microscope to elucidate anatomical differences. Stem and root bark exhibited characteristic medullary ray and secretary canal size. Leaf surface, petiole, and midrib exhibited characteristic inner morphologies including stomatal size, parenchyma, and epidermal cell diameter, as well as ratio of vascular bundle thickness to diameter among the samples. Orthogonal projections to latent structures discriminant analysis of anatomical data efficiently differentiated the five species. To evaluate chemical differences among the five species, we quantified (-)-catechin, (-)-catechin-7-O-ß-D-apiofuranoside, (-)-catechin-7-O-α-L-rhamnopyranoside, (-)-catechin-7-O-ß-D-xylopyranoside, (-)-catechin-7-O-ß-D-glucopyranoside, and (-)-catechin-5-O-ß-D-apiofuranoside using high-performance liquid chromatography with a diode-array detector. (-)-Catechin-7-O-ß-D-apiofuranoside content was the highest among all compounds in all species, and (-)-catechin-7-O-α-L-rhamnopyranoside content was characteristically the highest in Ulmus parvifolia among the five species. Overall, the Ulmus species tested was able to be clearly distinguished on the basis of anatomy and chemical composition, which may be used as scientific criteria for appropriate identification and standard establishment for commercialization of these species.

The changes of joint-position sense, peroneal strength, postural control, and functional performance ability after the modified Broström procedure for chronic ankle instability.

Purpose: Residual functional ankle instability regardless of the restoration of mechanical stability after the lateral ligament repair or reconstruction can cause recurrent sprain. The purpose of this study was to identify the sequential changes of joint-position sense, peroneal strength, postural control, and functional performance ability after the modified Broström procedure (MBP) for chronic ankle instability. Methods: A total of 46 patients (46 ankles) who underwent the MBP for chronic ankle instability were eligible for this study and were followed up for 1 year postoperatively. The changes of joint-position sense and peroneal strength were periodically evaluated with an isokinetic dynamometer. Postural control ability was evaluated using a one-leg stance test with eyes closed. The functional performance ability examination comprised a one-leg hop test, a six-meter hop test, and a cross three-meter hop test. Results: The error in joint-position sense significantly improved from a mean 4.3° to 2.8° (p < 0.001). Peak torque for eversion significantly improved from a mean 18.2 Nm to 21.2 Nm (p = 0.024). Balance retention time significantly improved from a mean 4.7 s to 6.4 s (p < 0.001). Among the functional performance tests, only the one-leg hop test showed a significant improvement postoperatively (p = 0.031). At 1 year postoperatively, the recovery ratios compared to the unaffected ankle were 67.9% in joint-position sense (p < 0.001), 86.9% in peroneal strength (p = 0.012), and 74.4% in postural control (p < 0.001), with significant side-to-side differences. Conclusion: Although joint-position sense, peroneal strength, postural control, and functional performance ability were significantly improved after the MBP, recovery ratios compared to the unaffected ankle were insufficient up to 1 year postoperatively. Level of Evidence: Level IV (prospective case series).

Overexpression of 4-hydroxyphenylpyruvate dioxygenase (IbHPPD) increases abiotic stress tolerance in transgenic sweetpotato plants.

Tocopherols are lipid-soluble compounds regarded as vitamin E compounds and they function as antioxidants in scavenging lipid peroxyl radicals and quenching reactive oxygen species (ROS). In our previous studies, we isolated five tocopherol biosynthesis genes from sweetpotato (Ipomoea batatas [L.] Lam) plants including 4-hydroxyphenylpyruvate dioxygenase (IbHPPD). HPPD is the first regulatory enzyme in vitamin E biosynthesis and serves to catalyze in the first steps α-tocopherol and plastoquinone biosynthesis by converting 4-hydroxyphenylpyruvate (HPP) to homogentisic acid (HGA). In this study, we generated transgenic sweetpotato plants overexpressing IbHPPD under the control of cauliflower mosaic virus (CaMV) 35S promoter (referred to as HP plants) via Agrobacterium-mediated transformation to understand the function of IbHPPD in sweetpotato. Three transgenic lines (HP3, HP14 and HP15) with high transcript levels of IbHPPD were selected for further characterization. Compared with non-transgenic (NT) plants, HP plants exhibited enhanced tolerance to multiple environmental stresses, including salt, drought, and oxidative stresses. In addition, HP plants showed increased tolerance to the herbicide sulcotrione, which is involved in the inhibition of the HPPD. Interestingly, after stress treatments, HP plants also showed higher abscisic acid (ABA) contents than NT plants. Under dehydrated condition, HP plants displayed an elevated α-tocopherol content to 19-27% in leaves compared with NT plants. These results indicate that increased abiotic stress tolerance in HP plants is related to inducing enhancement of α-tocopherol and ABA contents.

Dibenzocyclooctadiene Lignans in Plant Parts and Fermented Beverages of Schisandra chinensis.

The fruit of Schisandra chinensis, Omija, is a well-known traditional medicine used as an anti-tussive and anti-diarrhea agent, with various biological activities derived from the dibenzocyclooctadiene-type lignans. A high-pressure liquid chromatography-diode array detector (HPLC-DAD) method was used to determine seven lignans (schisandrol A and B, tigloylgomisin H, angeloylgomisin H, schisandrin A, B, and C) in the different plant parts and beverages of the fruit of S. chinensis grown in Korea. The contents of these lignans in the plant parts descended in the following order: seeds, flowers, leaves, pulp, and stems. The total lignan content in Omija beverages fermented with white sugar for 12 months increased by 2.6-fold. Omija was fermented for 12 months with white sugar, brown sugar, and oligosaccharide/white sugar (1:1, w/w). The total lignan content in Omija fermented with oligosaccharide/white sugar was approximately 1.2- and 1.7-fold higher than those fermented with white sugar and brown sugar, respectively. A drink prepared by immersion of the fruit in alcohol had a higher total lignan content than these fermented beverages. This is the first report documenting the quantitative changes in dibenzocyclooctadiene-type lignans over a fermentation period and the effects of the fermentable sugars on this eco-friendly fermentation process.

Overexpression of the Golden SNP-Carrying Orange Gene Enhances Carotenoid Accumulation and Heat Stress Tolerance in Sweetpotato Plants.

Carotenoids function as photosynthetic accessory pigments, antioxidants, and vitamin A precursors. We recently showed that transgenic sweetpotato calli overexpressing the mutant sweetpotato (Ipomoea batatas [L.] Lam) Orange gene (IbOr-R96H), which carries a single nucleotide polymorphism responsible for Arg to His substitution at amino acid position 96, exhibited dramatically higher carotenoid content and abiotic stress tolerance than calli overexpressing the wild-type IbOr gene (IbOr-WT). In this study, we generated transgenic sweetpotato plants overexpressing IbOr-R96H under the control of the cauliflower mosaic virus (CaMV) 35S promoter via Agrobacterium-mediated transformation. The total carotenoid contents of IbOr-R96H storage roots (light-orange flesh) and IbOr-WT storage roots (light-yellow flesh) were 5.4-19.6 and 3.2-fold higher, respectively, than those of non-transgenic (NT) storage roots (white flesh). The ß-carotene content of IbOr-R96H storage roots was up to 186.2-fold higher than that of NT storage roots. In addition, IbOr-R96H plants showed greater tolerance to heat stress (47 °C) than NT and IbOr-WT plants, possibly because of higher DPPH radical scavenging activity and ABA contents. These results indicate that IbOr-R96H is a promising strategy for developing new sweetpotato cultivars with improved carotenoid contents and heat stress tolerance.

Anti-obesity activity of anthocyanin and carotenoid extracts from color-fleshed sweet potatoes.

The anti-obesity effects of anthocyanin and carotenoid extracts from color-fleshed potatoes were studied with 3T3-L1 cells in vitro and high-fat diet (HFD)-induced obese mice in vivo. Treatment of 3T3-L1 adipocytes with anthocyanin and carotenoid extracts, respectively, after differentiation induction significantly inhibited fat accumulation by 63.1 and 83.5%. Studies of adipogenesis inhibition showed that the anthocyanin extract acts at intermediate stages, whereas the carotenoid extract influences all the stages. The extracts significantly diminished triglyceride (TG) content and peroxisome proliferator-activated receptor gamma (PPARγ) protein expression during adipogenesis of the intermediate stage. Oral administration of anthocyanin and carotenoid extracts, respectively, to HFD-fed mice significantly reduced weight gain and restored TG levels to normal or lower as compared to the HFD-fed group with improvement of a lipid profile, TG to HDL-C ratio. Histological differences in liver tissues revealed that the extracts protected the liver tissue from adipogenesis by HFD fed. This research presents the first direct demonstration that the two pigment extracts from sweet potato exhibit anti-obesity activities. PRACTICAL APPLICATIONS: Anthocyanins and carotenoids are the main pigments of purple- and orange-fleshed sweet potatoes, respectively, which are highly nutritious foods with antidiabetic and antioxidant properties. Obesity is a rapidly growing health problem that increases major risk factors of several serious diseases including cardiovascular diseases, diabetes, and cancer. The results of this research suggest that anthocyanin and carotenoid-rich extracts from color-fleshed sweet potatoes may be useful as supplementary ingredients for the treatment of obesity and related diseases.

Development and characterization of a superabsorbing hydrogel film containing Ulmus davidiana var. Japonica root bark and pullulan for skin wound healing.

Ulmus davidiana var. japonica (UD) has widely been used in Korean traditional medicine for the treatment of various types of diseases including inflammation and skin wounds. The UD root bark powders possess gelling activity with an excellent capacity for absorbing water. This distinct property could make the UD root bark powders to be a great material for manufacturing a gel film specifically for the healing of large and highly exudating wounds (e.g., pressure sores and diabetic ulcers). In this research, we separated the UD root bark powder into 4 different samples based on their sizes and then tested their water absorption capacity and flowability. Based on these results, 75-150 µm sized and below 75 µm sized samples of UD root bark powders were chosen, and UD gel films were prepared. The UD gel films showed good thermal stability and mechanically improved properties compared with pullulan only gel film with excellent swelling capacity and favorable skin adhesiveness. Further, in the animal studies with the skin wound mice model, the UD gel films exhibited significant therapeutic effects on accelerating wound closure and dermal regeneration. Overall, this study demonstrated the applicability of UD root bark powders for hydrogel wound dressing materials, and the potential of UD gel films to be superior wound dressings to currently available ones.

Comparative Studies of Fraxinus Species from Korea Using Microscopic Characterization, Phytochemical Analysis, and Anti-Lipase Enzyme Activity.

Fraxinus species belongs to the Oleaceae family, commonly known as Ash tree, and has been utilized as a folk medicine with various medicinal properties, including anti-obesity activity. The goal of the present study was to establish quality control parameters using microscopic characterization, phytochemical differentiation, and anti-lipase activity evaluation of five Fraxinus plants in Korea. Microscopic evaluation of the lower surface, petiole, and midrib of leaves, and stem bark showed discriminative anatomical characteristics, such as the stomatal index of the lower leaf surface; the number of sclerenchyma cells, and the diameter of parenchyma cells in the petiole and midrib; and the cork cell size and fiber frequency in the stem bark. Phytochemical analysis using high-performance liquid chromatography revealed the significant variation in the chemical profiles of the 12 major secondary metabolites among the samples. The orthogonal projections to latent structure-discrimination analysis efficiently differentiated each group belonging to each Fraxinus plant with the anatomical and quantification data. F. rhynchophylla and ligstroside showed the most potent anti-lipase activity among the plants and the 12 major metabolites, respectively. These findings could serve as the scientific criteria for the appropriate identification and establishment of standards for the use of Fraxinus species as medicinal plants.

Comparative Inner Morphological and Chemical Studies on Reynoutria Species in Korea.

Reynoutria species are medicinal plants that belong to the family Polygonaceae and are widely distributed in eastern Asia, North America and Europe. Although the phylogeny and morphological and anatomical studies of some species in Korea have been previously reported, there are no discriminative anatomical and chemical data available. Therefore, anatomical characterization of the leaf, stem and root, and high performance liquid chromatography-diode array detector (HPLC-DAD) analyses were carried out to assess the differences in anatomical and chemical profiles among the Reynoutria plants in Korea, i.e., R. japonica, R. sachalinensis, R. forbesii and R. japonica for. elata. The anatomical evaluation showed discriminative characteristics, such as the shape of the stomata and the stomatal index of the lower leaf surface; the ratio of the adaxial/abaxial height, the size of the vascular bundles and the frequency of druse in the midrib, petiole, and stem; and the pericycle number in the root. For the HPLC analysis, ten compounds corresponding to each major peak were isolated from R. japonica roots and their structures were identified by comprehensive spectroscopic studies. Samples collected before the flowering season showed higher contents of these ten major compounds than those collected after the flowering season. The orthogonal projections to latent structures-discrimination analysis (OPLS-DA) with the inner morphological and HPLC quantification results, clearly discriminated these plants. These results provide anatomical parameters and HPLC profiling that can be used to distinguish the four Reynoutria plants, which supports quality control for their precise identification.

Anatomical characterization and LC-MS profiling of Adenophora roots from Korea

ABSTRACT "Sasam (沙參)" is a crude drug that is defined in the in Korean Herbal Pharmacopoeia as the root of Adenophora triphylla var. japonica (Regel.) Hara or A. stricta Miq., Campanulaceae. The dried roots of the Adenophora spp. are available in markets, and the roots of various species are similar to each other in shape, making it difficult to distinguish one from another using only the outer morphological appearance. Therefore, the present study aimed to establish quality control parameters for pharmacognostic evaluation and differentiation of five Adenophora species and two varieties grown in Korea. Inner morphological evaluation of the root of these plants was accomplished and preliminary chemical analysis was performed by liquid chromatography-mass spectrometry profiling. As a result, significant differences among samples were found in anatomical characteristics such as number and thickness of cork layer, existence of stone cell in cork layer, frequency of vessels, and area of intercellular space. Significant differences were found among the samples in the content of three components including shashenoside I and a new alkyl glycoside, adenophoroside I. These findings could provide the scientific criteria for the proper identification and establishment of standards for the use of "Sasam".

Transgenic sweetpotato plants overexpressing tocopherol cyclase display enhanced α-tocopherol content and abiotic stress tolerance.

Oxidative stress caused by reactive oxygen species (ROS) under various environmental stresses significantly reduces plant productivity. Tocopherols (collectively known as vitamin E) are a group of lipophilic antioxidants that protect cellular components against oxidative stress. Previously, we isolated five tocopherol biosynthesis genes from sweetpotato (Ipomoea batatas [L.] Lam) plants, including tocopherol cyclase (IbTC). In this study, we generated transgenic sweetpotato plants overexpressing IbTC under the control of cauliflower mosaic virus (CaMV) 35S promoter (referred to as TC plants) via Agrobacterium-mediated transformation to understand the function of IbTC in sweetpotato. Three transgenic lines (TC2, TC9, and TC11) with high transcript levels of IbTC were selected for further characterization. High performance liquid chromatography (HPLC) analysis revealed that α-tocopherol was the most predominant form of tocopherol in sweetpotato tissues. The content of α-tocopherol was 1.6-3.3-fold higher in TC leaves than in non-transgenic (NT) leaves. No significant difference was observed in the tocopherol content of storage roots between TC and NT plants. Additionally, compared with NT plants, TC plants showed enhanced tolerance to multiple environmental stresses, including salt, drought, and oxidative stresses, and showed consistently higher levels of photosystem II activity and chlorophyll content, indicating abiotic stress tolerance. These results suggest IbTC as a strong candidate gene for the development of sweetpotato cultivars with increased α-tocopherol levels and enhanced abiotic stress tolerance.

A single amino acid change at position 96 (Arg to His) of the sweetpotato Orange protein leads to carotenoid overaccumulation.

KEY MESSAGE: IbOr-R96H resulted in carotenoid overaccumulation and enhanced abiotic stress tolerance in transgenic sweetpotato calli. The Orange (Or) protein is involved in the regulation of carotenoid accumulation and tolerance to various environmental stresses. Sweetpotato IbOr, with strong holdase chaperone activity, protects a key enzyme, phytoene synthase (PSY), in the carotenoid biosynthetic pathway and stabilizes a photosynthetic component, oxygen-evolving enhancer protein 2-1 (PsbP), under heat and oxidative stresses in plants. Previous studies of various plant species demonstrated that a single-nucleotide polymorphism (SNP) from Arg to His in Or protein promote a high level of carotenoid accumulation. Here, we showed that the substitution of a single amino acid at position 96 (Arg to His) of wild-type IbOr (referred to as IbOr-R96H) dramatically increases carotenoid accumulation. Sweetpotato calli overexpressing IbOr-WT or IbOr-Ins exhibited 1.8- or 4.3-fold higher carotenoid contents than those of the white-fleshed sweetpotato Yulmi (Ym) calli, and IbOr-R96H overexpression substantially increased carotenoid accumulation by up to 23-fold in sweetpotato calli. In particular, IbOr-R96H transgenic calli contained 88.4-fold higher levels of ß-carotene than those in Ym calli. Expression levels of carotenogenesis-related genes were significantly increased in IbOr-R96H transgenic calli. Interestingly, transgenic calli overexpressing IbOr-R96H showed increased tolerance to salt and heat stresses, with similar levels of malondialdehyde to those in calli expressing IbOr-WT or IbOr-Ins. These results suggested that IbOr-R96H is a useful target for the generation of efficient industrial plants, including sweetpotato, to cope with growing food demand and climate change by enabling sustainable agriculture on marginal lands.

Chemical constituents of the root bark of Ulmus davidiana var. japonica and their potential biological activities.

The root bark of Ulmus davidiana var. japonica (Ulmaceae), commonly known as yugeunpi, has been used as a traditional Korean medicine for the treatment of gastroenteric and inflammatory disorders. As part of continuing projects to discover bioactive natural products from traditional medicinal plants with pharmacological potential, phytochemical investigation of the root bark of this plant was carried out. This led to the successful isolation of a new chromane derivative (1) and 22 known compounds: catechin derivatives (2-5), megastigmane glycoside (6), dihydrochalcone glycosides (7 and 8), flavanone glycosides (9 and 10), coumarins (11 and 12), lignan derivatives (13-17), and phenolic compounds (18-23). The structure of the new compound (1) was determined with 1D and 2D NMR spectroscopy and HR-ESIMS, and its absolute configurations were achieved by chemical reactions and the gauge-including atomic orbital (GIAO) NMR chemical shifts calculations. All the isolated compounds were evaluated for their potential biological activities including neuro-protective, anti-neuroinflammatory, and anti-Helicobacter pylori activities. Among the isolates, compounds 1, 8, and 20 displayed stronger potency by causing a greater increase in the production and the activity of nerve growth factor (NGF) in C6 glioma cells (147.04 ±â€¯4.87, 206.27 ±â€¯6.70, and 143.70 ±â€¯0.88%, respectively), whereas compounds 11, 14, and 19 inhibited nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated murine microglial cells (IC50 of 18.72, 12.31, and, 21.40 µM, respectively). In addition, compounds 1, 11, 18, and 20 showed anti-H. pylori activity with MIC values of 25 or 50 µM against two strains of H. pylori 51 and 43504. These findings provide scientific evidence that supports the traditional usage of U. davidiana var. japonica root bark in the treatment of gastroenteric and inflammatory disorders.

A new anti-Helicobacter pylori juglone from Reynoutria japonica.

A 70% ethanol extract from the root portion of Reynoutria japonica afforded one new and three known juglone derivatives, namely, 2-methoxy-6-acetyl-7-methyljuglone (1), 2-ethoxy-6-acetyl-7-methyljuglone (2), 2-methoxy-7-acetonyljuglone (3), and 3-acetyl-7-methoxy-2-methyljuglone (4) together with two phenolics (5 and 6), an anthraquinone (7), a stilbene (8) and a phthalide (9). Their structures were elucidated on the basis of comprehensive spectroscopic studies including IR, MS, and 1H, 13C, 2D NMR spectra. Compound 3 is a new compound in nature, and compounds 4-6 have been isolated for the first time from R. japonica. The isolates were evaluated for their antibacterial activity against three strains (43504, 51, and 26695) of Helicobacter pylori. The four isolated juglone derivatives (1-4) showed potent growth inhibitory activity. Among them, compounds 1-3 exhibited stronger inhibitory activity than those of the positive controls, juglone and metronidazole, for the three strains and that of another reference, clarithromycin, for the 43504 and 51 strains. Specifically, the new juglone compound 3 displayed the most potent antibacterial activity against all three strains, 43504, 51, and 26695, with MIC values of 0.06, 0.06 and 0.13 µM, respectively, and MIC50 values of 0.14, 0.11 and 0.15 µM, respectively.

Down-regulation of lycopene ε-cyclase expression in transgenic sweetpotato plants increases the carotenoid content and tolerance to abiotic stress.

Carotenoids are required for many biological processes in plants and humans. Lycopene ε-cyclase (LCY-ε) catalyzes the conversion of lycopene into lutein via the α-branch carotenoid biosynthesis pathway. Down-regulation of IbLCY-ε by RNAi increases carotenoid accumulation and salt stress tolerance in transgenic sweetpotato calli. As the role of IbLCY-ε in carotenoid biosynthesis and environmental stress responses in whole plants is poorly understood, transgenic sweetpotato (RLE plants) with reduced expression of IbLCY-ε were developed. RLE plants contained higher levels of total carotenoid and ß-carotene, due to an elevated ß-carotene/lutein ratio rather than increased de novo biosynthesis. RLE plants showed high reactive oxygen species/radical-scavenging activity. They also exhibited an enhanced tolerance of both salt and drought stress, which was associated with lower membrane permeability and a higher photosynthetic rate, respectively. Elevated carotenoid accumulation in RLE plants mitigated the reductions in leaf photosystem II efficiency and chlorophyll induced by abiotic stress. Expression of the carotenoid cleavage genes 9-cis-epoxycarotenoid dioxygenase, carotenoid cleavage dioxygenase 1 (CCD1) and CCD4 was higher in RLE plants, as was abscisic acid accumulation. IbLCY-ε silencing thus offers an effective approach for developing sweetpotato plants with increased tolerance to abiotic stress that will grow on global marginal lands with no reduction in nutritional value.