Enhanced Cell Growth of Adipocyte-Derived Mesenchymal Stem Cells Using Chemically-Defined Serum-Free Media.

The multipotency and anti-inflammatory effects of mesenchymal stem cells (MSCs) make them attractive for cell therapy in regenerative medicine. A large number of MSCs is required for efficient therapy owing to the low homing efficiency of MSCs to target sites. Furthermore, owing to limitations in obtaining sufficient amounts of MSCs, in vitro expansion of MSCs that preserves their differentiation and proliferative potential is essential. The animal factor included in culture media also limits clinical application. In this study, adipose-derived MSCs showed a significantly higher proliferation rate in STK2, a chemically-defined medium, than in DMEM/FBS. The expression of MSC surface markers was increased in the culture using STK2 compared to that using DMEM/FBS. Tri-lineage differentiation analyses showed that MSCs cultured in STK2 were superior to those cultured in DMEM/FBS. In addition, MSCs cultured in STK2 showed a reduced senescence rate, small and homogenous cell size, and were more genetically stable compared to those cultured in DMEM/FBS. Furthermore, secretome analysis showed that the expression of factors related to proliferation/migration, anti-inflammation, and differentiation were increased in STK2 culture medium compared to DMEM/FBS. Taken together, these results suggest that culture using STK2 medium offers many advantages through which it is possible to obtain safer, superior, and larger numbers of MSCs.

Oligochitosan as a potential anti-acne vulgaris agent: combined antibacterial effects against Propionibacterium acnes.

To develop an antibacterial treatment for acne vulgaris using natural substance with few side effects, we investigated the antibacterial activities of oligochitosan against acne-related bacteria, particularly Propionibacterium acnes. Oligochitosan showed potent antibacterial effect on P. acnes. Especially, 10 kDa oligochitosan presented the highest antimicrobial effect with minimum inhibitory concentration values of 32-64 µg/mL on P. acnes. In addition, oligochitosan clearly reversed the antibacterial effect of tetracycline and erythromycin on P. acnes in the combination mode. The combination of tetracycline- or erythromycine-10 kDa oligochitosan resulted in a median ΣFIC range of 0.02-0.56, suggesting that the antibiotics-oligochitosan combination resulted in an antibacterial synergy against P. acnes. Thus, the results obtained in this research strongly supported that erythromycin and tetracycline will restore the antibacterial activity against P. acnes in the combination mode with 10 kDa oligochitosan.

Studies on antimicrobial activity of Poncirus trifoliata ethyl extract fraction against methicillin-resistant Staphylococcus aureus and to elucidate its antibacterial mechanism.

Traditional medicinal plants contain a wide variety of chemicals that have potent antibacterial activity. To find an alternative agent of overcoming the problems of methicillin-resistant Staphylococcus aureus (MRSA), the antibacterial mechanism of Ponciruss trifoliata against MRSA was investigated. Ethyl acetate (EtOAc)-soluble extract of P. trifoliata methanolic extract was evaluated for antibacterial activity using minimum inhibitory concentration (MIC). An EtOAc sub-fraction 08 (EA08) from silica-gel open column chromatography exhibited strong anti-MRSA activity. Apart from the study to isolate single compound from EA08, a synergistic antibacterial effect between the sub-fraction and ß-lactam antibiotics against MRSA was determined. In order to elucidate the antibacterial restoring mechanism of EA08 on MRSA, mRNA expression of mecA gene and production penicillin-binding protein 2a (PBP2a) encoded by mecA gene were monitored. EA 08 showed the strongest antibacterial activity with MIC value of 256 µg ml(-1). MIC of oxacillin against MRSA was dramatically reduced from 512 to 16 µg ml(-1) in combination with 256 µg ml(-1) of EA08. The fractional inhibitory concentration index of oxacillin was measured at 0.53 in combination with EA08 against MRSA, suggesting that EA08-oxacillin combinations exert synergetic effect against MRSA. The analysis of RT-PCR and Western blotting profiles revealed that EA08 inhibited mRNA expression of mecA gene and production PBP2a, which is a key determinant for ß-lactam antibiotic resistance, in a dose-dependent manner. These results indicated that EA08 eventually led to the reduction or inhibition of PBP2a production through translational inhibition in MRSA.

The mechanism of antibacterial activity of phlorofucofuroeckol-A against methicillin-resistant Staphylococcus aureus.

To find more effective ways of overcoming methicillin-resistant Staphylococcus aureus (MRSA), there has been considerable interest in the use of marine-derived constituents as alternatives to control pathogenic microorganisms. In this study, we investigated whether phlorofucofuroeckol-A (PFF) isolated from the edible brown alga Eisenia bicyclis suppressed production or function of penicillin-binding protein 2a (PBP2a). The antimicrobial mode of action of PFF in MRSA was identified by measuring cell membrane integrity and using the time-kill curve method. We attempted to determine the antimicrobial effects of PFF on the expression level of the resistance determinants mecA and its regulatory genes mecI and mecR1 in MRSA by reverse transcriptase polymerase chain reaction. PFF suppressed mecI, mecR1, and mecA gene expression in a dose-dependent manner. In addition, we revealed PFF mediates the suppressive effect of PBP2a expression in MRSA by Western blot analysis. PFF suppressed production of the PBP2a protein, suggesting that PFF probably acts by controlling the methicillin resistance-associated genes involved in the cell wall and production of PBP2a. These results demonstrate that PFF isolated from E. bicyclis significantly suppressed the expression of the methicillin resistance-associated genes and production of PBP2a, which is considered the primary cause of methicillin resistance.

Highly dispersed ruthenium nanoparticle-embedded mesoporous silica as a catalyst for the production of gamma-butyrolactone from succinic anhydride.

In this study, a novel, strategic method was developed for the synthesis of a mesoporous silica catalyst embedded with ruthenium nanoparticles (RuNPs/SiO2) by combining the polyol and modified sol-gel methods. By applying this new procedure, uniformly synthesized ruthenium nanoparticles with an average size of 3.8 nm and 95% spherical shape were highly dispersed in the mesoporous silica support material. Coordinated carbonyl groups of PVP remaining from the synthesis of the RuNPs were effectively removed by the thermal treatment (calcined at 573 K for 4 h) and the sythesized RuNPs/SiO2 catalysts were reduced under hydrogen at 20 bar for 2 h. These catalysts were analyzed using transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR), N2 adsorption-desorption, and X-ray diffraction (XRD). After the thermal treatment and the reduction procedure, the size and shape of the embedded RuNPs were nearly unchanged, and the catalyst was active in the liquid-phase hydrogenation of succinic anhydride (SAN) to selectively form y-butyrolactone (GBL) with a maximum yield of 90.1%. This novel catalyst preparation is a potentially useful method for the synthesis of metal nanoparticles as heterogeneous catalysts.

In vitro and in vivo immunostimulatory effects of hot water extracts from the leaves of Artemisia princeps Pampanini cv. Sajabal.

ETHNOPHARMACOLOGICAL RELEVANCE: Artemisia princeps Pampanini (Asteraceae) is used as a traditional medicine to immune function-related diseases, such as dysmenorrhea, inflammation, cancer, and ulcers. AIM OF THE STUDY: The purpose of this study is to evaluate the immunostimulatory effects of the hot water extract from the leaves of Artemisia princeps Pampanini (WAPP) in recombinant interferon-γ (rIFN-γ)-primed RAW 264.7 macrophages and in cyclophosphamide (20mg/kg, i.p.)-induced immunosuppressed Sprague-Dawley rats. MATERIALS AND METHODS: RAW 264.7 macrophages were treated with WAPP and production and expressions of nitric oxide (NO) and tumor necrosis factor-α (TNF-α) via nuclear factor-κB (NF-κB) were detected by immunoassay, western blot, qRT-PCR and reporter gene assay. In addition, in vivo immunomodulatory activity was studied by cyclophosphamide-induced myelosuppression in rats. RESULTS: In rIFN-γ-primed RAW 264.7 macrophages, pretreatment with WAPP increased the productions of nitric oxide (NO) and tumor necrosis factor-α (TNF-α),and increased the expressions of inducible nitric oxide synthase (iNOS) at the protein level and of iNOS and TNF-α at the mRNA level. Molecular data revealed that WAPP upregulated the transcriptional activity and translocation of nuclear factor-κB (NF-κB) by activating inhibitory kappa B-α (IκB-α) degradation and phosphorylation. Furthermore, WAPP upregulated the phosphorylations of p38 MAP kinase, c-Jun NH2-terminal kinase (JNK), and extracellular signal-regulated kinase 1/2 (ERK1/2). In cycloheximide-induced immunosuppressed rats, pretreatment with WAPP (100, 200, or 400mg/kg, p.o.) increased the serum levels of albumin and globulin, and reduced immobility times. CONCLUSION: Our results suggest that upregulations of the expressions of iNOS and TNF-α via the activations of NF-κB and MAPK are responsible for the immunostimulatory effects of WAPP.

Anti-methicillin-resistant Staphylococcus aureus (MRSA) substance from the marine bacterium Pseudomonas sp. UJ-6.

A multivalent approach to discover a novel antibiotic substance against methicillin-resistant Staphylococcus aureus (MRSA), a marine bacterium, UJ-6, exhibiting an antibacterial activity against MRSA was isolated from seawater. The isolated strain was identified to be Pseudomonas sp. by the morphology, biochemical, and genetical analyses. The ethyl acetate extract of Pseudomonas sp. UJ-6 culture showed significant ant-MRSA activity. Bioassay-guided isolation of the extract using a growth inhibitory assay led to the isolation and identification of an active compound exhibiting anti-MRSA activity. Based on the analyses of the physicochemical and spectroscopic data including nuclear magnetic resonance and mass, the compound was identified to be 1-acetyl-beta-carboline. The minimum inhibitory concentration (MIC) of the compound was determined to be in a range of 32-128 µg/ml against MRSA strains. The MIC values against MRSA were superior or equal to those of other natural compounds such as catechins, suggesting that 1-acetyl-beta-carboline would be a good candidate in applications of the treatment of MRSA infection.

Pancreatic lipase inhibitory activity of phlorotannins isolated from Eisenia bicyclis.

Pancreatic lipase is a potential therapeutic target for the treatment of diet-induced obesity in humans. In an ongoing search for new pancreatic lipase inhibitors from natural sources, a methanolic extract of marine brown algae, Eisenia bicyclis, showed a significant inhibitory effect against pancreatic lipase. Bioassay-guided isolation of this methanolic extract using a pancreatic lipase inhibitory assay led to the isolation and identification of six known phlorotannins: eckol (1), fucofuroeckol A (2), 7-phloroeckol (3), dioxindehydroeckol (4), phlorofucofuroeckol A (5), and dieckol (6). The structures were established on the basis of nuclear magnetic resonance and mass spectrometry spectroscopic data interpretation. Among the isolated phloroglucinol polymers, compounds 2 and 3 showed potent inhibitory effects on pancreatic lipase with IC50 values ranging from 37.2 ± 2.3 to 12.7 ± 1.0 µM, respectively.

In vitro antibacterial activity and synergistic antibiotic effects of phlorotannins isolated from Eisenia bicyclis against methicillin-resistant Staphylococcus aureus.

Six phlorotannins, isolated from Eisenia bicyclis, were evaluated for antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA). The minimum inhibitory concentrations (MIC) of the compounds were in the range 32 to 64 µg/mL. Phlorofucofuroeckol-A (PFF) exhibited the highest anti-MRSA activity, with an MIC of 32 µg/mL. An investigation of the interaction between these compounds and the ß-lactam antibiotics ampicillin, penicillin, and oxacillin revealed synergistic action against MRSA in combination with compound PFF. To our knowledge, this is the first report on the anti-MRSA activity of phlorotannins from E. bicyclis. The results obtained in this study suggest that the compounds derived from E. bicyclis can be a good source of natural antibacterial agents against MRSA.

Removal of off-flavors from sea tangle (Laminaria japonica) extract by fermentation with Aspergillus oryzae.

This study was conducted to reduce off-flavors from sea tangle (Laminaria japonica) extract since the typical odor of seaweed is an impediment to consumption of seaweed products. Fifty-six odor compounds emitted from the extract were identified using a thermal desorbor and gas chromatography-mass spectrometry. On the basis of expected odor intensity (EOI), isovaleric acid (41.2%), allyl isothiocyanate (22.6%), octanal (11.1%) and acetaldehyde (10.6%) were responsible for about 85.5% of total odor strength in the extract, suggesting that the odor of the extract might mainly be attributed to these compounds. The EOIs of odor compounds were dramatically reduced or disappeared during fermentation by Aspergillus oryzae. Total EOI of the odor compounds decreased about four-fold, compared with that of the raw extract (control) after the four days of fermentation at 30°C with agitation at 120 rpm. These results indicate that microbial fermentation may be a good strategy to reduce off-flavors of seaweeds.

α-Glucosidase- and α-amylase-inhibitory activities of phlorotannins from Eisenia bicyclis.

BACKGROUND: In an effort to develop alternative therapeutic agents, strong inhibitory activity against α-glucosidase and α-amylase was detected in Eisenia bicyclis methanolic extract. RESULTS: In this study, two phlorotannins were isolated from E. bicyclis and characterised by chromatography and nuclear magnetic resonance. The active substances were identified as fucofuroeckol A (FF) and dioxinodehydroeckol (DD). To the authors' knowledge, this is the first report of the identification of these substances in E. bicyclis. However, to date, no antidiabetic activity of FF and DD has been reported. Both phlorotannins demonstrated significant inhibitory activity against α-glucosidase and α-amylase. FF showed potent antidiabetic activity, with IC(50) values of 131.34 nmol L(-1) against α-glucosidase and 42.91 µmol L(-1) against α-amylase. The corresponding IC(50) values of DD were 93.33 nmol L(-1) and 472.7 µmol L(-1) . Furthermore, kinetic analysis revealed that FF and DD exhibited non-competitive inhibitory activity against α-glucosidase. CONCLUSION: These results suggest that FF and DD may be candidates for the development of an antidiabetic pharmaceutical agent or food additive.

Protective effect of fermented sea tangle against ethanol and carbon tetrachloride-induced hepatic damage in Sprague-Dawley rats.

Sea tangle has long been used as Korean folk remedy to promote material health, and is one of the popular dietary supplement. This study was designed to evaluate the protective effect of fermented sea tangle (FST) against ethanol and carbon tetrachloride (CCl(4))-induced hepatotoxicity in rats. Sprague-Dawley rats were orally treated with FST (25, 250, 2500 mg/kg/day) with administration of ethanol (5 mL/kg) for 13 weeks and the single intraperitoneal (i.p.) dose of 50% CCl(4) (5 mL/kg/day, CCl(4) in olive oil) at 12 week, and repeated i.p. dose of 20% CCl(4) (2 mL/kg/day) for 1 week. Hepatotoxicity was evaluated by measuring the serum levels of glutamic pyruvate transaminase (GPT), gamma glutamyl transpeptidase (gamma-GT) and malondialdehyde (MDA) as well as the tissue levels of antioxidant enzyme such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). Ethanol and CCl(4)-induced the rat liver damage, and significantly increased (p<0.05) the GPT, gamma-GT and MDA levels, and decreased the SOD, CAT and GPx levels. However, treatment with FST could decrease serum GPT, gamma-GT, and MDA levels significantly in plasma, and increase the activities of SOD, CAT, and GPx in liver tissues compared with ethanol and CCl(4)-treated group.

Synergistic effects between aminoethyl-chitosans and beta-lactams against methicillin-resistant Staphylococcus aureus (MRSA).

Two kinds of aminoethyl-chitosans (AEC), AEC90 and AEC50, which had degrees of deacetylation of 90% and 50%, respectively, were prepared and their synergistic effects in combination with beta-lactams including ampicillin, penicillin, and oxacillin against two standard methicillin-resistant Staphylococcus aureus (MRSA) strains and twelve clinical isolated MRSA strains were investigated. When AECs and beta-lactams were combined, synergistic effects were observed with fractional inhibitory concentration (FIC) indices of 0.252-0.508, and the MICs of beta-lactams in the presence of AECs were dramatically reduced.

Antibacterial activity of aminoderivatized chitosans against methicillin-resistant Staphylococcus aureus (MRSA).

This work describes the anti-MRSA activity of aminoderivatized chitosans. Two kinds of aminoethyl-chitosans (AEC), AEC90 and AEC50, having degrees of deacetylation of 90% and 50%, respectively, exhibited the strongest anti-MRSA activities by presenting MICs of 16-64 microg/mL against two standard strains and twelve clinical isolates. The bactericidal activity, thermal and pH stability, and cell membrane integrity effects of AEC90 and AEC50 are also discussed.

Isolation and characterization of a marine algicidal bacterium against the harmful raphidophyceae Chattonella marina.

A bacterial strain named AB-4 showing algicidal activity against Chattonella marina was isolated from coastal water of ULjin, Republic of Korea. The isolated strain was identified as Bacillus sp. by culture morphology, biochemical reactions, and homology research based on 16S rDNA. The bacterial culture led to the lysis of algal cells, suggesting that the isolated strain produced a latent algal-lytic compound. Amongst changes in algicidal activity by different culture filtrate volumes, the 10% (100 microl/ml) concentration showed the biggest change in algicidal activity; there, estimated algicidal activity was 95%. The swimming movements of Chattonella marina cells were inhibited because of treatment of the bacterial culture; subsequently, Chattonella marina cells became swollen and rounded. With longer exposure time, algal cells were disrupted and cellular components lost their integrity and decomposed. The released algicide(s) were heat-tolerant and stable in pH variations, except pH 3, 4, and 5. Culture filtrate of Bacillus sp. AB-4 was toxic against harmful algae bloom (HAB) species and nontoxic against livefood organisms. Bacillus sp. AB-4 showed comparatively strong activity against Akashiwo sanguinea, Fibriocapsa japonica, Heterosigma akashiwo, and Scrippsiella trochoidea. These results suggest that the algicidal activity of Bacillus sp. AB-4 is potentially useful for controlling outbreaks of Chattonella marina.

[The effect of a health diary program in solitary elderly].

PURPOSE: This study was to develop and to evaluate a health diary program for solitary elderly. METHODS: The research design was a nonequivalent control group pretest-posttest design. There were 30 subjects the experimental group and 54 in the control group among solitary elderly over age 65. The independent variable was the health diary program, and the dependent variables were perceived health status, ADL, IADL, Blood pressure, blood sugar, quality of life, self efficacy and health promoting behaviors. The health diary program was performed for 50 minutes, twice a week for 8 weeks. Data was collected from April 3 to June 23, 2006. RESULTS: The experimental group showed a significant difference in perceived health status, blood sugar, ADL, IADL, quality of life, self efficacy, and health promotion behaviors than the control group. There were no significant differences in systolic & diastolic blood pressure. CONCLUSION: A health diary program showed good effects on improving health status, quality of life, self efficacy, and health promotion behaviors. Therefore, we recommend this program be utilized as a health promoting program for the solitary elderly in the community.

Relation of urinary incontinence, menopausal symptoms, and life satisfaction in middle-age Korean women.

Associations of urinary incontinence, menopausal symptoms and life satisfaction of 235 middle-age Korean women (40 to 59 years) were investigated, based on responses to a self-report questionnaire. Urinary incontinence was positively correlated with menopausal symptoms (r =.24) and negatively correlated with life satisfaction (r=-.11). Life satisfaction was negatively correlated with menopausal symptoms (r = -.28). All rs indicate weak associations and account for small common variables. Even so, results suggest effective treatment of urinary incontinence and menopausal symptoms could enhance life satisfaction through promoting healthy behavior and emotional health in such middle-age women.

Perceptions, knowledge and misuse of an oriental herbal drug: a survey of 608 Korean female nursing college students.

This study was designed to survey college students about their knowledge and use of Uwhangchungsimwon (UC), a Traditional Oriental herbal remedy for emergency and acute treatment of stroke, numbness, hypertension, epilepsy and arteriosclerosis. Students at three female nursing colleges in Korea completed a questionnaire containing 29 questions about the students' demographics, knowledge and use of UC, and the placebo effect. Of 608 respondents, 217 (36%) had taken UC. Eighty-three per cent reported that UC alleviated examination tension and 28% knew that it is used as an emergency drug for stroke. Forty-two per cent believed that UC has a "placebo effect" and 20% considered its effects were more than that of a placebo. Fifty-eight per cent of those using UC felt that it was effective in inducing psychological relaxation, 4% considered that it increased their energy levels, and 6% felt it was effective in "clearing the head"; 33% reported no effects. Users also reported adverse effects: 30% reported drowsiness, 17% reported a "heavy head" and 15% experienced indigestion. Most respondents (73%) obtained information about UC from family members and friends and 33% from newspapers or the media. These results confirm health practitioner should recognize that UC is used widely for purposes other than its intended purpose (i.e., to treat stroke and hypertension) and that such use can cause adverse effects. More systematic research is needed to increase the safety of consumers when using herbs and herbal supplements.

Histological grading of IgA nephropathy predicting renal outcome: revisiting H. S. Lee's glomerular grading system.

BACKGROUND: The glomerular grading system is useful to compare biopsy specimens and to predict the natural course of disease in IgA nephropathy (IgAN), although no grading system can be perfect. METHODS: H. S. Lee's grading system for IgAN was refined as follows: grade I, normal or focal mesangial cell proliferation; grade II, diffuse mesangial cell proliferation, or <25% of glomeruli with crescent (Cr)/segmental sclerosis (SS)/global sclerosis (GS); grade III, 25-49% of glomeruli with Cr/SS/GS; grade IV, 50-75% of glomeruli with Cr/SS/GS; grade V, >75% of glomeruli with Cr/SS/GS. This refined H. S. Lee grading system was then tested for clinical relevance on 187 patients with IgAN followed up for an average of 6.5 years (minimum, 3 years). In the survival analysis, a modified primary end-point (progressive renal disease) was used. RESULTS: The glomerular grades were significantly related to hypertension, serum creatinine levels and the amounts of proteinuria at time of biopsy. By univariate analysis, glomerular grades, hypertension, renal insufficiency and significant proteinuria (> or =1 g/day) were significantly associated with progressive renal disease. By multivariate analysis using the Cox regression model, glomerular grades, renal insufficiency and significant proteinuria were independent prognostic factors for progressive renal disease. At the end of follow-up, glomerular grades were significantly related to serum creatinine levels, amounts of proteinuria, hypertension and progressive renal disease. CONCLUSIONS: These findings indicate that the refined H. S. Lee grading system for IgAN is useful in assessing the patients' clinical outcome and is sufficiently simple and easy to reproduce as to be universally applicable in prognostic work.

Impact of qigong exercise on self-efficacy and other cognitive perceptual variables in patients with essential hypertension.

OBJECTIVES: The purpose of this study was to investigate the impact of practicing qigong on middle-age subjects with essential hypertension. Impacts on blood pressure, reported self-efficacy, perceived benefit, and emotion were observed. DESIGN: Thirty-six (36) adult volunteers were assigned to either a waiting list control or a qigong group that practiced two 30-minute qigong programs per week over 8 consecutive weeks. RESULTS: Systolic and diastolic blood pressure was significantly reduced in members of the qigong group after 8 weeks of exercise. Significant improvements in self-efficacy and other cognitive perceptual efficacy variables were also documented in the qigong group compared to the original situation described above. CONCLUSIONS: This pilot study demonstrates the positive effects of practicing qigong on controlling blood pressure and enhancing perceptions of self-efficacy.