Structures of three ependymin-related proteins suggest their function as a hydrophobic molecule binder

Ependymin was first discovered as a predominant protein in brain extracellular fluid in fish and was suggested to be involved in functions mostly related to learning and memory. Orthologous proteins to ependymin called ependymin-related proteins (EPDRs) have been found to exist in various tissues from sea urchins to humans, yet their functional role remains to be revealed. In this study, the structures of EPDR1 from frog, mouse and human were determined and analyzed. All of the EPDR1s fold into a dimer using a monomeric subunit that is mostly made up of two stacking antiparallel beta-sheets with a curvature on one side, resulting in the formation of a deep hydrophobic pocket. All six of the cysteine residues in the monomeric subunit participate in the formation of three intramolecular disulfide bonds. Other interesting features of EPDR1 include two asparagine residues with glycosylation and a Ca2+-binding site. The EPDR1 fold is very similar to the folds of bacterial VioE and LolA/LolB, which also use a similar hydrophobic pocket for their respective functions as a hydrophobic substrate-binding enzyme and a lipoprotein carrier, respectively. A further fatty-acid binding assay using EPDR1 suggests that it indeed binds to fatty acids, presumably via this pocket. Additional interactome analysis of EPDR1 showed that EPDR1 interacts with insulin-like growth factor 2 receptor and flotillin proteins, which are known to be involved in protein and vesicle translocation

Accumulation of Phenolic Compounds and Glucosinolates in Sprouts of Pale Green and Purple Kohlrabi (Brassica oleracea var. gongylodes) under Light and Dark Conditions

Kohlrabi is considered an important dietary vegetable worldwide. In this study, we investigated the growth and accumulation of phenolic compounds (PCs) and glucosinolates in sprouts of pale green and purple kohlrabi (Brassica oleracea var. gongylodes) in response to light and dark conditions. Pale green kohlrabi presented high fresh weight and root length irrespective of light treatment, whereas under dark conditions, it presented higher fresh weight and shoot length than purple kohlrabi. In contrast, the root length of both kohlrabies increased markedly under light conditions compared to that under dark conditions. Thirteen PCs and eight glucosinolates were detected and quantified in 10-day-old pale green and purple kohlrabies. In both kohlrabies, the individual and total phenolic levels were much higher under the light treatment than under the dark treatment. Under light and dark conditions, the total phenolic content was 6362.13 and 5475.04 µg/g dry weight in the pale green kohlrabi, respectively, whereas in the purple kohlrabi, it was 10,115.76 and 9361.74 µg/g dry weight, respectively. Dark conditions favored higher accumulation of glucosinolates than light conditions. Progoitrin, neoglucobrassicin, glucoerucin, and 4-methoxyglucobrassicin were the predominant glucosinolates in both kohlrabies and were present in much higher amounts in the pale green kohlrabi. In pale green kohlrabi under dark conditions, the total glucosinolates content was 4.75 and 2.62 times higher than that of the purple kohlrabi under light and dark conditions, respectively. Among individual glucosinolates, in the pale green kohlrabi under the dark condition, progoitrin was found to have the highest content, which was 90.28 and 54.51 times higher than that in the purple kohlrabi under light and dark conditions, respectively. These results show that the phenolic and glucosinolates levels varied widely, and these variations between the two types of kohlrabi under both light and dark conditions were significant. Our findings suggest that light and dark conditions enhance the accumulation of PCs and glucosinolates, respectively, during the development of kohlrabi seedlings

Effects of Chilling Treatment on Baicalin, Baicalein, and Wogonin Biosynthesis in <i>Scutellaria baicalensis</i> Plantlets

When plants are exposed to stressful conditions, they modulate their nutrient balance by regulating their primary and secondary metabolisms to adapt. In this study, changes in primary and secondary metabolites elicited by chilling stress treatment and the effects of treatment duration were examined in roots of Scutellaria baicalensis (S. baicalensis) plantlets. The concentrations of most sugars (maltose, glucose, sucrose, and fructose) and of several amino acids (proline and GABA), which are crucial regarding plant defense mechanisms, increased with increasing duration of chilling stress. Furthermore, salicylic acid levels increased after two-day chilling treatments, which may enhance plant tolerance to cold temperatures. The concentrations of flavones (baicalin, baicalein, and wogonin) increased during chilling stress, and those of phenolic acids (ferulic acid and sinapic acid) increased after two-day chilling treatments. The concentrations of these flavones were positively correlated with sucrose levels which acted as energy sources

Comprehensive histologic analysis of interstitial lipolysis with the 1444 nm wavelength during a 3-month follow-up

A number of near-infrared wavelengths have been proposed and studied for laser lipolysis, but the histologic evaluation of tissue response to laser lipolysis during long-term follow-up has been lacking. A 1444 nm Nd:YAG laser with better absorption in both fat and water has recently attracted attention. The present study was designed to investigate the comprehensive histopathology of 1444 nm Nd:YAG laser-assisted lipolysis at different energy levels during a 3-month follow-up. Laser lipolysis was performed on porcine fat tissue in vivo using a 1444 nm Nd:YAG laser (AccuSculpt®, Lutronic Corporation, Ilsan, Republic of Korea) and the total energies delivered interstitially to 10x10 cm2 areas were 750 J, 1500 J, 2250 J, 3000 J, 3750 J, 4500 J, and 5250 J. Biopsy samples were taken and histologically analyzed immediately after biopsy and at 1, 2, 4, and 12 weeks postoperatively. With a fluence setting above 3000J/100 cm2, inflammation was severe and remained by the 3-month follow-up, resulting in severe scarring of the fat tissue. Below this energy level, mild lobular inflammation in the early phase biopsy had resolved with no scarring by the 3-month follow-up. No histologic changes in the epidermis or dermal connective tissue were present. This study suggested that controlling the energy level is important for clinical applications of laser lipolysis with no significant complications

Effects of Carbohydrates on Rosmarinic Acid Production and In Vitro Antimicrobial Activities in Hairy Root Cultures of Agastache rugosa

Agastache rugosa (popularly known as Korean mint) belongs to the Lamiaceae family and comprises 22 species of perennial aromatic medicinal species native to East Asian countries, such as Korea, Taiwan, Japan, and China. A. rugosa contains many phenolic compounds that exhibit pharmacological and physiological activities, including antioxidant, anticancer, antiviral, antifungal, and antibacterial activities. The highest concentrations of rosmarinic acid and its isomers have been reported in the roots of A. rugosa. In this in vitro study, hairy roots of A. rugosa were obtained and the carbohydrates (sorbitol, mannitol, glucose, maltose, galactose, mannose, and sucrose) were evaluated to determine those that were optimal for rosmarinic acid production and hairy root growth. Antioxidant and antibacterial activities of extracts of A. rugosa were also assessed. The best carbon source for A. rugosa hairy root cultures was sucrose, considering biomass productivity (0.460 &plusmn; 0.034 mg/30 mL), rosmarinic acid production (7.656 &plusmn; 0.407 mg/g dry weight), and total phenolic content (12.714 &plusmn; 0.202 mg/g gallic acid equivalent). Antioxidant and antimicrobial activities were displayed by A. rugosa hairy roots cultured in liquid medium supplemented with 100 mM sucrose. Twenty-five bacterial strains, including multidrug-resistant bacteria and one pathogenic yeast strain, were used for antimicrobial screening of A. rugosa hairy roots. The hairy root extracts displayed antibacterial activity against Micrococcus luteus (KCTC 3063) and Bacillus cereus (KCTC 3624). The inhibition of these bacteria was greater using A. rugosa hairy roots with the highest levels of phenolic compounds cultured in the presence of sucrose, compared to hairy roots with the lowest levels of phenolic compounds cultured in the presence of fructose. Considering hairy root biomass, phenolic compound production, and antibacterial activity, sucrose is the best carbon source for A. rugosa hairy root cultures

Effects of Carbohydrates on Rosmarinic Acid Production and In Vitro Antimicrobial Activities in Hairy Root Cultures of <i>Agastache rugosa</i>

Agastache rugosa (popularly known as Korean mint) belongs to the Lamiaceae family and comprises 22 species of perennial aromatic medicinal species native to East Asian countries, such as Korea, Taiwan, Japan, and China. A. rugosa contains many phenolic compounds that exhibit pharmacological and physiological activities, including antioxidant, anticancer, antiviral, antifungal, and antibacterial activities. The highest concentrations of rosmarinic acid and its isomers have been reported in the roots of A. rugosa. In this in vitro study, hairy roots of A. rugosa were obtained and the carbohydrates (sorbitol, mannitol, glucose, maltose, galactose, mannose, and sucrose) were evaluated to determine those that were optimal for rosmarinic acid production and hairy root growth. Antioxidant and antibacterial activities of extracts of A. rugosa were also assessed. The best carbon source for A. rugosa hairy root cultures was sucrose, considering biomass productivity (0.460 ± 0.034 mg/30 mL), rosmarinic acid production (7.656 ± 0.407 mg/g dry weight), and total phenolic content (12.714 ± 0.202 mg/g gallic acid equivalent). Antioxidant and antimicrobial activities were displayed by A. rugosa hairy roots cultured in liquid medium supplemented with 100 mM sucrose. Twenty-five bacterial strains, including multidrug-resistant bacteria and one pathogenic yeast strain, were used for antimicrobial screening of A. rugosa hairy roots. The hairy root extracts displayed antibacterial activity against Micrococcus luteus (KCTC 3063) and Bacillus cereus (KCTC 3624). The inhibition of these bacteria was greater using A. rugosa hairy roots with the highest levels of phenolic compounds cultured in the presence of sucrose, compared to hairy roots with the lowest levels of phenolic compounds cultured in the presence of fructose. Considering hairy root biomass, phenolic compound production, and antibacterial activity, sucrose is the best carbon source for A. rugosa hairy root cultures

RT-PCR analysis of osteoblast-specific genes expression during 2 weeks of culture in osteogenically differentiated hTMSCs.

<p>Bone morphogenetic protein-2 (BMP-2), runt-related transcription factor 2 (Runx2), bone sialoprotein (BSP), type I collagen (Col1), osteocalcin (OC), osterix (Osx), and osteopontin (OP) mRNA expression were analyzed. The experiment was performed in triplicate for each sample. Differences among the groups were evaluated by one-way analysis of variance (ANOVA; <i>p</i><0.05). Except for OP, there were no significant differences in expression levels among the groups.</p

Effect of donor age on the osteogenic differentiation potential of hTMSCs.

<p>Cells were cultured in osteogenic induction medium. Cells aggregated, formed nodules, and accumulated calcium deposits over a 2-week period. A–D: Alkaline phosphatase staining of hTMSCs cultured from each age group before osteogenic induction. E–H: Alkaline phosphatase staining of hTMSCs cultured from each age group after osteogenic induction. Alkaline phosphatase activity, which is indicative of osteoblastic differentiation, is shown as red staining. I–L: Alizarin red staining of hTMSCs cultured from each age group after osteogenic induction. (A, E, and I: group I; B, F, and J: group II; C, G, and K: group III; D, H, and L: group IV). Alizarin Red staining was used to detect precipitated calcium salt, which is a marker of differentiation. Scale bars: 100 µm. Visual assessment data demonstrated that hTMSCs cultured from donors of all age groups showed the alkaline phosphatase activity increase after osteogenic induction and similar levels of alkaline phosphatase and Alizarin Red staining.</p

Proliferation of hTMSCs according to age group.

<p>Cellular proliferation was monitored over a period of 14 days. hTMSCs from all groups exhibited rapid proliferation from day 2 to 4; hTMSCs from group (II) expanded more rapidly than those from the other groups. However, there were no statistically significant differences in proliferation rate among the groups.</p

Effects of donor age on the immunophenotypic characteristics of hTMSCs.

<p>Effects of donor age on the immunophenotypic characteristics of hTMSCs.</p