Potent in vivo antifungal activity against powdery mildews of pregnane glycosides from the roots of Cynanchum wilfordii.

Two new pregnane glycosides, kidjoranine 3-O-ß-D-glucopyranosyl-(1 → 4)-ß-D-glucopyranosyl-(1 → 4)-α-L-cymaropyranosyl-(1 → 4)-ß-D-cymaropyranosyl-(1→4)-α-L-diginopyranosyl-(1 → 4)-ß-D-cymaropyranoside (5) and caudatin 3-O-ß-D-glucopyranosyl-(1 → 4)-ß-D-glucopyranosyl-(1 → 4)-α-L-cymaropyranosyl-(1 → 4)-ß-D-cymaropyranosyl-(1 → 4)-α-L-diginopyranosyl-(1 → 4)-ß-D-cymaropyranoside (6), were isolated from the roots of Cynanchum wilfordii along with four known compounds (1-4). The antifungal activities of the six compounds against barley powdery mildew caused by Blumeria graminis f. sp. hordei were compared to the antifungal activity of polyoxin B. The caudatin glycosides (1, 4, and 6) showed stronger antifungal activities than polyoxin B, whereas kidjoranine glycosides (2, 3, and 5) had weaker activities than polyoxin B. A wettable powder-type formulation (C. wilfordii-WP20) of the ethyl acetate extract from C. wilfordii roots prohibited the development of barley powdery mildew much more effectively than the commercial fungicide polyoxin B-WP10. In addition, C. wilfordii-WP20 effectively controlled strawberry powdery mildew caused by Sphaerotheca humuli under greenhouse conditions. Thus, the crude extract containing the pregnane glycosides can be used as a botanical fungicide for the environmentally benign control of powdery mildews.

Inhibitory effects of broccoli extract on Escherichia coli O157:H7 quorum sensing and in vivo virulence.

Broccoli extract (BE) has numerous beneficial effects on human health including anticancer activity. Quorum sensing (QS), mediated by self-produced autoinducer (AI) molecules, is a key process for the production of virulence determinants in pathogenic bacteria. BE suppressed AI-2 synthesis and AI-2-mediated bacterial motility in a dose-dependent manner in Escherichia coli O157:H7. In addition, expression of the ler gene that regulates AI-3 QS system was also diminished in response to treatment with BE. Furthermore, in an in vivo efficacy test using Caenorhabditis elegans as a host organism, C. elegans fed on E. coli O157:H7 in the presence of BE survived longer than those fed solely on the pathogenic bacteria. Quantitative real-time PCR analysis indicated that quercetin was the most active among the tested broccoli-derived compounds in downregulating virulence gene expression, while treatment with myricetin significantly suppressed the expression of the eae gene involved in type III secretion system. These data suggest that BE and its flavonoid constituents can inhibit expression of QS-associated genes, thereby downregulating the virulence attributes of E. coli O157:H7 both in vitro and in vivo. This study clearly elucidates BE's QS-inhibitory activity and suggests that BE has the potential to be developed as an anti-infective agent.

Neuroprotective effects of SG-168 against oxidative stress-induced apoptosis in PC12 cells.

Postmortem examinations of tissues of humans and rodents with a host of neurodegenerative conditions, including Alzheimer's and Parkinson's diseases, have identified oxidative damage in proteins, lipids, and DNA. The aim of this study was to better understand the cellular mechanisms of neuronal cell degeneration induced via oxidative stress and the protective roles of bioactive substance. In order to achieve this aim, we established a screening program to discover therapeutic agents that exhibit preferential neuroprotective activity in H(2)O(2)-treated PC12 cells. During the course of our screening program, we isolated an active compound, SG-168, from Dendrobium nobile Lindley and identified it as a neuroprotective agent. SG-168 was identified as a compound with an acetal skeleton, a prototypical compound, by electrospray ionization-mass spectrometry analysis and various nuclear magnetic resonance spectroscopic methods. The protective effect of SG-168 in PC12 cells with H(2)O(2)-induced oxidative damage was investigated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction assay. As expected, incubation with H(2)O(2) for 2 hours resulted in cell viability of 31.8% compared to the control, while pretreatment of SG-168 increased cell viability by 15-50% compared to the H(2)O(2)-stressed control cells. These results showed that SG-168 inhibits H(2)O(2)-induced apoptotic cell death. Interestingly, flow cytometric analysis showed that H(2)O(2)-treated PC12 cells incubated with SG-168 exhibited greatly suppressed apoptosis. In summation, the results of this study suggest that SG-168 has potential as a new antioxidant agent against neuronal diseases.

Selective cytotoxicity of Ponciri Fructus against glucose-deprived PANC-1 human pancreatic cancer cells via blocking activation of GRP78.

Pancreatic cancer cells are sometimes exposed to stressful microenvironments such as glucose deprivation, hypoxia, and starvation of other nutrients. These stresses, which are characteristic of poorly vascularized solid tumors, activate the unfolded protein response (UPR). The UPR is a stress-signaling pathway present in tumor cells that is associated with molecular chaperone GRP78. Induction of GRP78 has been found to increase cell survival and decrease apoptotic potential through genetic alterations. Thus GRP78 may represent a novel target in the development of anticancer drugs. Here we established a novel screening program to identify chaperone modulators that exhibit preferential cytotoxic activity in glucose-deprived pancreatic cancer cells. During the course of our screening, we isolated an active substance, Ponciri Fructus (PF), from an herbal medicine source and identified it as a down-regulator of GRP78. As expected, PF inhibited expression of the GRP78 protein under glucose-deprivation conditions in a dose-dependent manner. Furthermore, it induced selective cytotoxicity against glucose-deprived cancer cells; this effect was not observed under normal growth conditions. We also detected apoptotic bodies on Hoechst staining and attempted to determine whether PF-induced apoptosis involved caspase-3 activation. Our results suggest that the GRP78-inhibitory action of PF was dependent on strict hypoglycemic conditions and that it resulted in the selective death of glucose-deprived pancreatic cancer cells.

Neuroprotective activity of Viola mandshurica extracts on hydrogen peroxide-induced DNA damage and cell death in PC12 cells.

This study was conducted to examine the neuroprotective effects of acetone extracts from Viola mandshurica (VME). The effect of VME on hydrogen peroxide (H(2)O(2))-induced DNA damage in PC12 cells was evaluated by the comet assay where VME (100 and 250 microg/mL) was a dose-dependent inhibitor of DNA damage induced by 500 micromol/L of H(2)O(2). The protective effect of VME against H(2)O(2)-induced oxidative damage on PC12 cells was investigated by an MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] reduction assay and lactate dehydrogenase (LDH) release assays. After 3 h of cell exposure to 500 micromol/L of H(2)O(2), a marked reduction in cell survival was observed. However, the reduction was significantly prevented by 100 and 250 microg/mL of VME. H(2)O(2) also induced severe apoptosis of the PC12 cells, which was indicated by Hoechst 33342 staining. Interestingly, the H(2)O(2)-stressed PC12 cells that were incubated with 100 and 250 microg/mL of VME had greatly suppressed apoptosis. The results suggest that VME could be a new antioxidant candidate against neuronal diseases.

Comparison of a long-pulse Nd:YAG laser and a combined 585/1,064-nm laser for the treatment of acne scars: a randomized split-face clinical study.

BACKGROUND: Nonablative laser is gaining popularity because of the low risk of complications, especially in patients with darker skin. OBJECTIVE: To compare the efficacy and safety of a long-pulse neodymium-doped yttrium aluminium garnet (Nd:YAG) laser and a combined 585/1,064-nm laser for the treatment of acne scars. MATERIALS AND METHODS: Nineteen patients with mild to moderate atrophic acne scars received four long-pulse Nd:YAG laser or combined 585/1,064-nm laser treatment sessions at fortnightly intervals. Treatments were administered randomly in a split-face manner. RESULTS: Acne scars showed mild to moderate improvement, with significant Echelle d'évaluation clinique des cicatrices d'acné (ECCA) score reductions, after both treatments. Although intermodality differences were not significant, combined 585/1,064-nm laser was more effective for deep boxcar scars. In patients with combined 585/1,064-nm laser-treated sides that improved more than long-pulse Nd:YAG laser-treated sides, ECCA scores were significantly lower for combined 585/1,064-nm laser treatment. Histologic evaluations revealed significantly greater collagen deposition, although there was no significant difference between the two modalities. Patient satisfaction scores concurred with physicians' evaluations. CONCLUSION: Both lasers ameliorated acne scarring with minimal downtime. In light of this finding, optimal outcomes might be achieved when laser treatment types are chosen after considering individual scar type and response.

Comparison of a 585-nm pulsed dye laser and a 1064-nm Nd:YAG laser for the treatment of acne scars: A randomized split-face clinical study.

BACKGROUND: No studies have reported a comparison of the pulsed dye laser (PDL) and the 1064-nm long-pulsed neodymium:yttrium-aluminum-garnet (Nd:YAG) laser treatment of acne scars in the same patient. OBJECTIVE: To compare the efficacies of these two lasers in the treatment of acne scars. METHODS: Eighteen patients received 4 sessions of PDL or Nd:YAG laser at 2-week intervals in a randomized split-face manner. RESULTS: Both lasers induced notable and comparable improvement in the appearance of acne scars, particularly superficial scars, with significant reductions in the scores associated with the clinical evaluation scale for acne scarring (ECCA). Histologic evaluations revealed significant increases in collagen production and deposition following both lasers. Patient satisfaction scores concurred with these improvements. Ice-pick scars and boxcar scars tended to respond better to PDL and Nd:YAG lasers, respectively. LIMITATIONS: The number of subjects was small. CONCLUSIONS: Both lasers are effective modalities for the treatment of acne scars. Optimal outcomes might be achieved considering scar types and responses to a specific laser.

Protective effects of Paeonia lactiflora pall on hydrogen peroxide-induced apoptosis in PC12 cells.

This study was conducted to examine the antioxidative and neuroprotective effects of Paeonia lactiflora pall (PLE). Total phenolic content of PLE was 89.65 mg of gallic acid equivalent per gram of PLE. IC(50) values for reducing power, hydrogen peroxide scavenging activity, and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity were 297.57, 3.33, and 32.74 microg, respectively. The protective effect of PLE against H(2)O(2)-induced oxidative damage to PC12 cells was investigated by an 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) reduction assay and lactate dehydrogenase (LDH) release assay. After 2 h of cell exposure to 0.5 mM H(2)O(2), a marked reduction in cell survival was observed. However, this reduction was significantly prevented by 10-100 microg/ml of PLE. H(2)O(2) also induced severe apoptosis of the PC12 cells, which was indicated by a flow cytometric analysis. Interestingly, the H(2)O(2)-stressed PC12 cells that had been incubated with PLE had greatly suppressed apoptosis. The results suggest that PLE could be a candidate for a new antioxidant against neuronal diseases.

Methanolic extracts of Uncaria rhynchophylla induce cytotoxicity and apoptosis in HT-29 human colon carcinoma cells.

In this paper, we report the anticancer activities of Uncaria rhynchophylla extracts, a Rubiaceae plant native to China. Traditionally, Uncaria rhynchophylla has been used in the prevention and treatment of neurotoxicity. However, the cytotoxic activity of Uncaria rhynchophylla against human colon carcinoma cells has not, until now, been elucidated. We found that the methanolic extract of Uncaria rhynchophylla (URE) have cytotoxic effects on HT-29 cells. The URE showed highly cytotoxic effects via the MTT reduction assay, LDH release assay, and colony formation assay. As expected, URE inhibited the growth of HT-29 cells in a dose-dependent manner. In particular, the methanolic URE of the 500 microg/ml showed 15.8% inhibition against growth of HT-29 cells. It induced characteristic apoptotic effects in HT-29 cells, including chromatin condensation and sharking occurring 24 h when the cells were treated at a concentration of the 500 microg/ml. The activation of caspase-3 and the specific proteolytic cleavage of poly (ADP-ribose) polymerase were detected over the course of apoptosis induction. These results indicate that URE contains bioactive materials with strong activity, and is a potential chemotherapeutic agent candidate against HT-29 human colon carcinoma cells.

Treatment-seeking behaviors and related epidemiological features in Korean acne patients.

Little is known about the treatment-seeking behaviors of acne patients, especially Asian acne patients. This study was performed to obtain detailed information about the treatment-seeking behaviors in Korean acne patients. Patients who visited the dermatology departments at 17 university hospitals completed a self-administered questionnaire. Most patients obtained information about acne from doctors or the Internet. The most important criteria for selecting a treatment method or choosing a particular clinic were effectiveness and accessibility. Patients used traditional medicine, visited beauty clinics, drank more water, and used over-the-counter topical agents more frequently than they sought doctors during the worsening period. The degree of satisfaction in treatment was found to depend on the total cost of treatment, number of places visited, site affected by acne, and emotional stress. Those who had experienced a side effect tended to have been treated for longer, to have paid more for treatment, and to have an associated skin disease. Treatments prescribed by dermatology clinics had the lowest aggravating rate, although improvement rates for family medicine clinics were also fairly high. This is the first study to investigate in detail the demographic features and characteristics of the treatment-seeking behaviors of acne patients in Asia.

Methanolic extracts of Plocamium telfairiae induce cytotoxicity and caspase-dependent apoptosis in HT-29 human colon carcinoma cells.

Natural marine products have recently become the focus of increased research interest, due to their potential pharmacological activities. Therefore, we have screened 50 varieties of marine seaweed and determined that the methanolic extracts from Plocamium telfairiae (PTE) exhibited a cytotoxic effect against HT-29 human colon carcinoma cells. In this study, we report on the cytotoxic activity and mechanism of PTE-induced apoptosis in HT-29 cells. The treatment of HT-29 cells with various PTE concentrations resulted in the inhibition of growth and the induction of apoptosis in a dose-dependent manner, as determined by the results of a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction assay, a lactate dehydrogenase release assay, a morphological assay, and a colony formation assay. Interestingly, we also detected apoptotic bodies on Hoechst staining and attempted to determine whether the PTE-induced apoptosis involved the caspase pathway, using a caspase colorimetric assay. The activation of caspases-8, -9, -3, and -7 and the specific proteolytic cleavage of poly(ADP-ribose) polymerase were detected over the course of apoptosis induction. Our results showed that PTE may function as a chemopreventive and/or chemotherapeutic agent in colon carcinoma cells via the reduction of cell viability and the induction of apoptosis.