Lupeol-induced nitric oxide elicits apoptosis-like death within Escherichia coli in a DNA fragmentation-independent manner.

Lupeol is known to be plentiful in fruits or plant barks and has an antimicrobial effect, however, its mode of action(s) has yet to be determined. To elucidate lupeol generates nitric oxide (NO), which is recognized for possessing an antimicrobial activity, intracellular NO was measured in Escherichia coli using DAF-FM. Using the properties of NO passing through plasma membrane easily, increased malondialdehyde levels have shown that lupeol causes lipid peroxidation, and the resulting membrane depolarization was confirmed by DiBAC4(3). These data indicated that lupeol-induced NO is related to the destruction of bacterial membrane. Further study was performed to examine whether NO, known as a cell proliferation inhibitor, affects bacterial cell division. As a result, DAPI staining verified that lupeol promotes cell division arrest, and followed by early apoptosis is observed in Annexin V/PI double staining. Even though these apoptotic hallmarks appeared, the endonuclease failed to perform properly with supporting data of decreased intracellular Mg2+ and Ca2+ levels without DNA fragmentation, which is confirmed using a TUNEL assay. These findings indicated that lupeol-induced NO occurs DNA fragmentation-independent bacterial apoptosis-like death (ALD). Additionally, lupeol triggers DNA filamentation and morphological changes in response to DNA repair system called SOS system. In accordance with the fact that ALD deems to SOS response, and that the RecA is considered as a caspase-like protein, increase in caspase-like protein activation occurred in E. coli wild-type, and no ΔRecA mutant. In conclusion, these results demonstrated that the antibacterial mode of action(s) of lupeol is an ALD while generating NO.

Efficacy of Ethyl Acetate Fraction of Cordyceps militaris for Cancer-Related Fatigue in Blood Biochemical and 1H-Nuclear Magnetic Resonance Metabolomic Analyses.

This study investigated the adjuvant effects for anticancer and antifatigue of the combination of Cordyceps militaris extract with sorafenib. The 5 extracts of C militaris were obtained through hexane, chloroform, ethyl acetate, butanol, and water and were evaluated for anticancer growth activity. Among these extracts, ethyl acetate extract of C militaris showed the best tumor growth inhibitory activity and the adjuvant effects in combination with sorafenib. As a result of biochemical analysis with serum, the combination of ethyl acetate extract of C militaris with sorafenib showed the adjuvant effects both improving hepatic function and relieving cancer-related fatigue. In addition, 1H-nuclear magnetic resonance-based metabolic profiling in liver tissues showed that the change of metabolism by ethyl acetate extract of C militaris with sorafenib was related with serum fatigue biomarkers. Therefore, the combination strategy such as ethyl acetate extraction of C militaris with sorafenib constitutes a promising therapeutic strategy in hepatocellular carcinoma, via the inhibition of cancer growth, the enhancement of liver function, as well as the alleviation of cancer-related fatigue.

Low-fluenced erbium:yttrium-aluminium-garnet laser treatment in combination with broadband light pretreatment for various pigmentation disorders in Asian patients.

BACKGROUND: Low-fluenced 2940-nm erbium (Er):yttrium-aluminium-garnet (YAG) resurfacing elicits ablative photothermal tissue reactions confined to the uppermost parts of the epidermis. OBJECTIVE: To demonstrate the efficacy and safety of low-fluenced ablative Er:YAG laser treatment in combination with broadband light (BBL) pretreatment for various pigmentation disorders. METHODS: In total, 35 Korean patients with various pigmentation disorders were pretreated with BBL, and then, low-fluenced Er:YAG laser resurfacing was performed with a beam size of 4 mm and a fluence of 1.0-1.5 J/cm2 . RESULTS: An average of 1.1 ± 0.4 sessions of combined BBL and low-fluenced Er:YAG resurfacing treatment was delivered to the patients. Most post-Er:YAG scaling fell off spontaneously over 3-5 days, and most of the post-BBL crusting disappeared spontaneously over 5-7 days. At 2 months after final treatment, the mean global aesthetic improvement scale score for the clinical improvement of pigmentation lesions was estimated as 2.5 ± 0.8, and that for the improvement of overall skin tone, texture, and wrinkles was 2.8 ± 1.0. CONCLUSION: Our data demonstrated that post-BBL, low-fluenced Er:YAG laser resurfacing can be used to effectively treat various pigmentation disorders in Asian patients. Further improvements in overall skin tone, texture, and wrinkles were also achieved without major side effects.

Broadband light treatment using static operation and constant motion techniques for skin tightening in Asian patients.

BACKGROUND: Broadband light (BBL) devices irradiate photons of different wavelength to induce photothermal reactions on various aging-related chromophores. OBJECTIVES: To evaluate three BBL treatment settings for skin tightening in Asian patients. METHODS: A total of 27 patients underwent three sessions of BBL treatment via (1) an 800-nm cutoff filter using a static operation technique and a 695-nm cutoff filter using a constant motion technique (group 1, N = 9), (2) an 800-nm cutoff filter using a constant motion technique (group 2, N = 9), and (3) a 590-nm cutoff filter using a constant motion technique (group 3, N = 9). RESULTS: The patients in group 1 presented marked clinical improvements in zygomatic wrinkles, nasolabial folds, and marionette lines, with a median overall global aesthetic improvement scale (GAIS) score of 3. Meanwhile, patients in group 2 exhibited noticeable improvements in zygomatic wrinkles, nasolabial folds, perioral expression wrinkles at the cheek, and marionette lines, with a median GAIS score of 3. Patients in group 3 experienced improvement in skin tone and texture, zygomatic wrinkles, nasolabial folds, and marionette lines, with a median GAIS score of 2. CONCLUSIONS: Our data demonstrated that BBL treatment for nonablative, noninvasive skin tightening elicits satisfactory clinical outcomes.

Effects on skin of Stichopus japonicus viscera extracts detected with saponin including Holothurin A: Down-regulation of melanin synthesis and up-regulation of neocollagenesis mediated by ERK signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Stichopus japonicus (sea cucumber), edible traditional food in Asia, and its extracts are renowned for their wound healing, pain relieving, and cosmetic effects in traditional medicine. Holothurins, toxins isolated from sea cucumber, are thought to be active components for their beneficial effects. However, researchers have yet to outline specific mechanisms thereof. AIM OF THE STUDY: The present study was designed to evaluate the anti-melanogenic and anti-wrinkle properties of S. japonicus viscera extracts (VF) on the skin via in vitro and ex vivo experiments and to assess the anti-aging effects of S. japonicus viscera extracts in relation to known wound healing and cosmetic processes. MATERIALS AND METHODS: The viscera of live S. japonicus specimens were freeze dried and ground into a powder. Aqueous extracts were subsequently prepared from the concentrated powder using a water extraction method. To investigate the inhibitory effects of VF on melanogenesis, mushroom tyrosinase activity assay and melanin assay were performed on Melan-A cells. To further delineate the anti-melanogenic properties of VF, western blot analysis for tyrosinase, TRP-1, TRP-2, MITF, and ERK was conducted. Changes in collagen synthesis in human dermal fibroblast (HDF) were evaluated via CCK-8 assay and immunocytochemistry to determine the anti-wrinkle effects of VF. Finally, anti-aging properties were examined in a human skin equivalent ex vivo model. RESULTS: In Melan-A cells, VF treatment reduced melanin contents in a concentration-dependent manner. The anti-melanogenic effects of VF appeared to be due to enzymatic inhibition of tyrosinase. In CCK-8 assay, VF also significantly increased the viability of HDFs in a concentration-dependent manner. Immunoblot analysis revealed phosphorylation of ERK in HDFs treated with VF. In a human skin equivalent ex vivo model (Neoderm®-ED), VF treatment at a concentration of 50 µg/ml enhanced collagen type IV and Ki-67 expression and downregulated MMP-9 expression. CONCLUSION: This study demonstrated that aqueous extracts from S. japonicus viscera are effective whitening and anti-aging agents that stimulate ERK signaling to inhibit melanin synthesis and promote collagen synthesis.

Therapeutic efficacy and safety of a 1927-nm fractionated thulium laser on pattern hair loss: an evaluator-blinded, split-scalp study.

Laser- or light-assisted therapies have been used to improve the perifollicular environment by upregulating the expression of growth factors and signaling molecules for hair restoration. The aim of our study was to preclinically and clinically evaluate the therapeutic efficacy and safety of a 1927-nm fractionated thulium laser on pattern hair loss (PHL). An in vivo hairless mouse study and an in vivo human skin environmental scanning electron microscopy (ESEM) study were performed with different power and energy settings. Thereafter, an evaluator-blinded, split-scalp study was conducted to evaluate hair thickness and density in 10 PHL patients treated with 12 sessions of fractionated thulium laser treatment with or without post-laser treatment application of a growth factor-containing (GF) solution. In in vivo hairless mouse skin, inverted cone-shaped zones of thulium laser-induced tissue coagulation (LITC) were noted immediately after treatment in the epidermis and upper to mid-dermis without remarkable ablative tissue injury. The ESEM study revealed round to oval-shaped zones of non-ablative LITC on the surface of the stratum corneum of a human subject immediately after laser irradiation. In PHL patients, 12 sessions of thulium laser monotherapy at 1-week intervals resulted in significantly increased hair density and thickness. Post-laser treatment application of GF solution offered additional therapeutic efficacy by improving hair density and thickness on the split scalp. The use of a fractionated thulium laser with or without post-laser therapy application of GF solution to treat PHL elicited remarkable improvements in hair thickness and hair counts.

Inhibitory effects of Stichopus japonicus extract on melanogenesis of mouse cells via ERK phosphorylation.

Stichopus japonicus has been used as a folk medicine and as an ingredient in traditional food in East Asian countries. In recent years, the bioactive compounds found in S. japonicus have been reported to possess efficacy in wound healing and may be of potential use in the cosmeceutical, pharmaceutical and biomedical industries. Although the components and their functions require further investigation, S. japonicus extracts exhibit anti­inflammatory properties, and may be used for cancer prevention and treatment. Although several reports have examined different aspects of S. japo-nicus, the effects of S. japonicus extract on melanogenesis in the skin has not been reported to date. Therefore the present study aimed to investigate the effects of S. japonicus extract on melanogenesis. Treatment with a mixture of S. japonicus extracts (MSCE) reduced melanin synthesis and tyrosinase (TYR) activity in mouse melanocyte cells lines, B16F10 and Melan­A. In addition, MSCE treatment reduced the protein expression levels of TYR, tyrosinase­related protein­1 and tyrosinase­related protein­2. The reduced protein levels may be the result of decreased microphthalmia­associated transcription factor (MITF) expression, which is an important regulator of melanogenesis. The reduced expression level of MITF was associated with delayed phosphorylation of extracellular signal­regulated kinase (ERK) induced by MSCE treatment. A specific MEK inhibitor, PD98059, significantly blocked MSCE­mediated inhibition of melanin synthesis. In conclusion, these results indicate that MSCE may be useful as a potential skin­whitening compound in the skin medical industry.

Rapid sequential separation of essential oil compounds using continuous heart-cut multi-dimensional gas chromatography-mass spectrometry.

A method for separation and identification of peaks in essential oil samples based on rapid repetitive heart-cutting using multidimensional gas chromatography (MDGC)-mass spectrometry (MS) coupled with a cryotrapping interface is described. Lavender essential oil is analyzed by employing repetitive heart-cut intervals of 1.00 and 1.50 min, achieved in a parallel MDGC-MS/GC-FID experiment. The number of peaks that were detected in 1D GC operation above a given response threshold more than tripled when MDGC-MS employing the cryotrapping module method was used. In addition, MDGC-MS enabled detection of peaks that were not individually evident in 1D GC-MS, owing to effective deconvolution in time of previously overlapped peaks in 1D GC. Thus separation using the cryomodulation approach, without recourse to using deconvolution software, was possible. Peaks widths decreased by about 5-7-fold with the described method, peak capacity increased from about 9 per min to 60 per min, and greater sensitivity results. Repeatability of retention times for replicate analyses in the multidimensional mode was better than 0.02% RSD. The present study suggests that the described heart-cutting technique using MDGC-MS can be used for general improvement in separation and identification of volatile compounds.