Chamaecyparis obtusa Suppresses Virulence Genes in Streptococcus mutans.

Chamaecyparis obtusa (C. obtusa) is known to have antimicrobial effects and has been used as a medicinal plant and in forest bathing. This study aimed to evaluate the anticariogenic activity of essential oil of C. obtusa on Streptococcus mutans, which is one of the most important bacterial causes of dental caries and dental biofilm formation. Essential oil from C. obtusa was extracted, and its effect on bacterial growth, acid production, and biofilm formation was evaluated. C. obtusa essential oil exhibited concentration-dependent inhibition of bacterial growth over 0.025 mg/mL, with 99% inhibition at a concentration of 0.2 mg/mL. The bacterial biofilm formation and acid production were also significantly inhibited at the concentration greater than 0.025 mg/mL. The result of LIVE/DEAD® BacLight™ Bacterial Viability Kit showed a concentration-dependent bactericidal effect on S. mutans and almost all bacteria were dead over 0.8 mg/mL. Real-time PCR analysis showed that gene expression of some virulence factors such as brpA, gbpB, gtfC, and gtfD was also inhibited. In GC and GC-MS analysis, the major components were found to be α-terpinene (40.60%), bornyl acetate (12.45%), α-pinene (11.38%), ß-pinene (7.22%), ß-phellandrene (3.45%), and α-terpinolene (3.40%). These results show that C. obtusa essential oil has anticariogenic effect on S. mutans.

Artemisia princeps Inhibits Biofilm Formation and Virulence-Factor Expression of Antibiotic-Resistant Bacteria.

In this study, we used ethanol extract of A. princeps and investigated its antibacterial effects against MRSA. Ethanol extract of A. princeps significantly inhibited MRSA growth and organic acid production during glucose metabolism at concentrations greater than 1 mg/mL (P < 0.05). MRSA biofilm formation was observed using scanning electron microscopy (SEM) and safranin staining. A. princeps extract was found to inhibit MRSA biofilm formation at concentrations higher than 2 mg/mL significantly (P < 0.05). Bactericidal effects of the A. princeps were observed using confocal laser microscopy, which showed that A. princeps was bactericidal in a dose-dependent manner. Using real-time PCR, expression of mecA, an antibiotic-resistance gene of MRSA, was observed, along with that of sea, agrA, and sarA. A. princeps significantly inhibited mecA, sea, agrA, and sarA, mRNA expression at the concentrations greater than 1 mg/mL (P < 0.05). The phytochemical analysis of A. princeps showed a relatively high content of organic acids and glycosides. The results of this study suggest that the ethanol extract of A. princeps may inhibit proliferation, acid production, biofilm formation, and virulence gene expressions of MRSA, which may be related to organic acids and glycosides, the major components in the extract.

Chamaecyparis obtusa Essential Oil Inhibits Methicillin-Resistant Staphylococcus aureus Biofilm Formation and Expression of Virulence Factors.

The emergence of antibiotic-resistant bacteria has caused difficulty in treating infectious diseases. Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most commonly recognized antibiotic-resistant bacteria. Novel antibiotics are urgently required to treat these bacteria. Raw materials derived from natural sources can be used for the development of novel antibiotics, such as Chamaecyparis obtusa (C. obtusa), which has been traditionally used in treating asthmatic disease. In this study, the antibacterial activity of the essential oil (EO) extracted from C. obtusa leaves against MRSA was investigated. MRSA growth and acid production from glucose metabolism were inhibited at concentrations greater than 0.1 mg/mL C. obtusa EO. MRSA biofilm formation was observed using scanning electron microscopy and safranin staining. C. obtusa EO inhibited MRSA biofilm formation at concentrations greater than 0.1 mg/mL. Using real-time polymerase chain reaction, mRNA expression of virulence factor genes, sea, agrA, and sarA, was observed. agrA expression was inhibited with C. obtusa EO concentrations greater than 0.2 mg/mL, whereas inhibition of sea and sarA expression was also observed at a concentration of 0.3 mg/mL. C. obtusa EO was analyzed by gas chromatography (GC) and GC coupled for mass spectrometry, which identified 59 constituents, accounting to 98.99% of the total EO. These findings suggest that C. obtusa EO has antibacterial effects against MRSA, which might be associated with the major components of C. obtusa EO, such as sabinene (19.06%), α-terpinyl acetate (16.99%), bornyl acetate (10.48%), limonene (8.54%), elemol (7.47%), myrcene (5.86%), γ-terpinene (4.04%), and hibaene (3.01%).

Effects of pulsed electromagnetic field therapy on delayed-onset muscle soreness in biceps brachii.

OBJECTIVE: to compare the effects of pulsed electromagnetic field (PEMF) therapy and sham treatment on DOMS-related variables in elbow flexors at 24, 48 and 72 h after delayed onset muscle soreness (DOMS) induction exercise. DESIGN: randomized, double-blind, placebo-controlled study. SETTING: Yonsei University laboratory. PARTICIPANTS: In total, 30 healthy male college students. MAIN OUTCOME MEASURES: Muscle soreness, peak torque, median frequency (MDF) and electromechanical delay (EMD) during isometric contraction at 24, 48 and 72 h after DOMS induction exercise. RESULTS: Overall, the application of the PEMF was found to be effective in reducing the physiological deficits associated with DOMS, including improved recovery of perceived muscle soreness, MDF, and EMD during isometric contraction. Our results did not show that PEMF treatment was mechanically more effective for isometric peak torque generation compared to the sham group. CONCLUSION: this study indicates that PEMF may be useful as a modality to reduce DOMS symptoms. However, further well-designed experiments are required to determine optimal treatment dosage and duration, and to investigate the physiological and clinical mechanisms of PEMF on DOMS.

The discovery of aurora kinase inhibitor by multi-docking-based virtual screening.

We report the discovery of aurora kinase inhibitor using the fragment-based virtual screening by multi-docking strategy. Among a number of fragments collected from eMololecules, we found four fragment molecules showing potent activity (>50% at 100 µM) against aurora kinase. Based on the explored fragment scaffold, we selected two compounds in our synthesized library and validated the biological activity against Aurora kinase.

Effects of a lipid-encapsulated zinc oxide supplement on growth performance and intestinal morphology and digestive enzyme activities in weanling pigs.

This study compared the effects of varying lipid content and dietary concentration of a lipid-encapsulated (LE) ZnO product to those of native ZnO and thereby to find insights into optimal lipid coating and dosage of the Zn supplement. A total of 192 21-d-old weanling pigs were allotted to 48 pens, after which each six pens received a ZnO-free basal diet supplemented with 125 ppm ZnO (100 ppm Zn; BASAL), 2,500 ppm Zn as native ZnO (HIGH), or 100 or 200 ppm Zn as LE ZnO (LE-100 or LE-250) containing 8%, 10%, or 12% lipid [LE-8%, LE-10%, or LE-12%, respectively; 2 × 3 factorial arrangement within the LE-ZnO diets (LE-ALL)] for 14 d. Forty pigs were killed at the end for histological and biochemical examinations. None of ADG, ADFI, gain:feed, and fecal consistency score differed between the LE-ALL and either of the BASAL and HIGH groups. Hepatic and serum Zn concentrations were greater (p <0.05) in the HIGH vs. LE-ALL group, but did not differ between LE-ALL and BASAL, between LE-100 and -250, or among LE-8%, -10%, and -12% groups. Villus height (VH), crypt depth (CD), and the VH:CD ratio in the duodenum, jejunum, and ileum did not differ between the LE-ALL and either of the BASAL and HIGH groups, except for a greater CD in the duodenum in the LE-ALL vs. HIGH group. Additionally, VH and CD in the duodenum and VH:CD in the jejunum were greater in the LE-250 vs. LE-100 group. Specific activities of sucrase, maltase, and leucine aminopeptidase in these intestinal regions and those of amylase and trypsin in the pancreas were not influenced by the lipid content or dietary concentration of LE ZnO and also did not differ between the LE-ALL and either of the BASAL and HIGH groups, except for a greater pancreatic amylase activity in the former vs. HIGH group. In conclusion, the present results indicate that the LE ZnO, regardless of its lipid percentage or supplementation level examined in this study, has no significant effect on growth performance, fecal consistency, or digestive enzyme activities of weanling pigs under the experimental conditions.

Antibacterial Activity of Rhus javanica against Methicillin-Resistant Staphylococcus aureus.

In the present study, the leaves of Rhus javanica (R. javanica) were extracted with ethanol, and we investigated the antimicrobial activity of the ethanol extract of R. javanica against methicillin-resistant Staphylococcus aureus (MRSA). Control groups were treated with media containing 0.1% DMSO. The ethanol extract of R. javanica inhibited the growth of MRSA at concentrations ranging from 0.05 to 0.2 mg/mL and inhibited acid production at concentrations higher than 0.1 mg/mL (P < 0.05). MRSA biofilm formation was determined by scanning electron microscopy and safranin staining. The ethanol extract of R. javanica inhibited the formation of MRSA biofilms at concentrations higher than 0.05 mg/mL. In confocal laser scanning microscopy, high concentration (0.4-1.6 mg/mL) of R. javanica extract showed bactericidal effect in a dose-dependent manner. In real-time PCR analysis, R. javanica extract showed the inhibition of the genetic expression of virulence factors such as mecA, sea, agrA, and sarA in MRSA. Preliminary phytochemical analysis revealed the strong presence of phenolics. These results suggest that R. javanica may be a useful medicinal plant for inhibiting MRSA, which may be related to the presence of phenolics in the R. javanica extract.

Kaurenoic Acid from Aralia continentalis Inhibits Biofilm Formation of Streptococcus mutans.

We isolated a single chemical compound from A. continentalis and identified it to be kaurenoic acid (KA) and investigated the influence of anticariogenic properties. Inhibitory effects of KA on cariogenic properties such as growth, acid production, biofilm formation, and the adherence of S. mutans were evaluated. Furthermore, real-time PCR analysis was performed to evaluate the influence of KA on the genetic expression of virulence factors. KA significantly inhibited the growth and acid production of S. mutans at 2-4 µ g/mL and 4 µ g/mL of KA, respectively. Furthermore, the adherence onto S-HAs was inhibited at 3-4 µ g/mL of KA and biofilm formation was significantly inhibited when treated with 3 µ g/mL KA and completely inhibited at 4 µ g/mL. Also, the inhibitory effect of KA on biofilm formation was confirmed by SEM. In confocal laser scanning microscopy, bacterial viability gradually decreased by KA in a dose dependent manner. Real-time PCR analysis showed that the expressions of gtfB, gtfC, gbpB, spaP, brpA, relA, and vicR were significantly decreased in S. mutans when it was treated with KA. These results suggest that KA from A. continentalis may be a useful agent for inhibiting the cariogenic properties of S. mutans.