Bifunctional mechanisms of autophagy and apoptosis regulations in melanoma from Bacillus subtilis natto fermentation extract.

Melanoma is one of the most dangerous malignant epidermal cancers. Natto freeze-drying extract (NFDE) and natto water extract (NWE) were isolated from natto, soybeans fermented by Bacillus subtilis natto, which were assessed as potential anti-melanoma agents. Cell cytotoxicity assays revealed significant anti-melanoma effects of NFDE and NWE in a dose-dependent manner, and exhibited low influences on normal skin cells, including Hs68, HaCaT and adipose tissue-derived stem cells (ADSCs), respectively. Through a flow cytometer assay and autophagy acridine orange staining, the cellular death phenomenon shifted from autophagy to apoptosis with the increased dosages. Reactive oxygen species (ROS) were enhanced using DCFDA (2,7-dichlorodihydrofluorescein diacetate) staining when melanoma cells were treated with the extract. NFDE and NWE treatments increase the oxidative stress of cancer cells and cause apoptosis by inhibiting AMP-activated protein kinase (AMPK). NFDE and NWE were considered to play a critical role in cell death through ROS adjustment, autophagy regulation and apoptosis promotion.

Purified Astaxanthin from Haematococcus pluvialis Promotes Tissue Regeneration by Reducing Oxidative Stress and the Secretion of Collagen In Vitro and In Vivo.

Intracellular reactive apoptosis and reactive oxygen species (ROS) play a crucial role in ultraviolet- (UV-) induced inflammation and aging reaction in human dermal tissues. This study determines the mechanism by which Haematococcus pluvialis extracts (HPE) and purified astaxanthin (HPA) to promote skin regeneration in the injured tissue in vitro and in vivo. The results show that HPE and HPA decrease the DNA damage and promote the secretion of collagen from the human normal fibroblast cell line (Hs68) in a dose-dependent manner. UV irradiation and HPA reduce oxidative stress damage due to phorbol-12-myristate-13-acetate (PMA). When skin cells are injured by free radicals, cells undergo a programmed cellular death. Cellular apoptotic death is determined using annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double staining to verify that there is no cell membrane asymmetry and that the nuclear membrane is broken. Inflammatory symptoms and apoptotic injuries to experimental rats in a group that is treated with HPA treated are decreased in a dose-dependent manner after UVB exposure (300 mJ/cm2) for 15 min in vivo, compared to the vehicle control group. These positive results show that HPA repairs UVB-triggered skin tissue injury and aging by conducting electrons out of cells to maintain a low level of oxidative stress so that collagen is synthesized in vitro and in vivo.

Antioxidant Graphene Oxide Nanoribbon as a Novel Whitening Agent Inhibits Microphthalmia-Associated Transcription Factor-Related Melanogenesis Mechanism.

In the melanin synthesis process, oxidative reactions play an essential role, and it is a good strategy to inhibit melanin production by reducing oxidative stress. Fullerene and its derivatives, or the complexes, were considered as strong free-radical scavengers, and we further applied multilayered sp2 nanocarbons to discover melanin synthesis inhibitory mechanisms. In the present study, we used novel nanomaterials, such as multiwalled carbon nanotubes (MWCNTs), short-type MWCNTs, graphene oxide nanoribbons (GONRs), and short-type GONRs, as anti-oxidative agents to regulate melanin production. The results showed that GONRs had better anti-oxidative capabilities in intracellular and extracellular oxidative stress analysis platforms than others. We proposed that GONRs have oxygen-containing functional groups. In the 2',7'-dichlorodihydrofluorescein diacetate assay, we found out GONR could chelate metal ions to scavenge reactive oxygen species. In the molecular insight view, we observed that these nanomaterials downregulated the melanin synthesis by decreasing microphthalmia-associated transcription factor-related gene expressions, and there were similar consequences in protein expressions. To sum up, GONRs is a potential agent as a novel antioxidant and skin-whitening cosmetology material.

Metabolic engineering probiotic yeast produces 3S, 3'S-astaxanthin to inhibit B16F10 metastasis.

3S, 3'S-Astaxanthin is the most powerful antioxidant to scavenge free radicals in the world. In this study, a 3S, 3'S-astaxanthin biosynthesis pathway was constructed in a probiotic yeast, Kluveromyces marxianus, denoted YEAST, and its bioactive metabolites were extracted for biofunctional assessments. The bio-safety examination was achieved by two animal models as following: First, no significant toxic effects on YEAST groups were found in zebrafish; Second, after feeding YEAST for 4 weeks, the rat-groups showed no visible abnormality, and no significant change of the body weight and blood biochemistry tests. The inhibition of lung metastasis of melanoma cells and the increment of the survival rate were demonstrated by feeding YEAST and injecting the intravenous commercial astaxanthin in vivo rodent model. Based on in vitro assays of 1,1-diphenyl-2-picryl-hydrazyl (DPPH) scavenging analysis, ferrous ion chelating ability, reducing power assessment, and mushroom tyrosinase inhibition evaluation, YEAST-astaxanthin showed anti-oxidative and tyrosinase suppressive properties. Taken together, the 3S, 3'S-astaxanthin producing probiotic yeast is safe to be used in the bio-synthesis of functional and pharmaceutical compounds, which have broad industrial applications on cosmetic, food and feed additive and healthcare.

Functional Analysis of Macromolecular Polysaccharides: Whitening, Moisturizing, Anti-Oxidant, and Cell Proliferation.

In this research we utilized extracts from two different nature products, Achatina fulica and Heimiella retispora, to enhance skin moisturizing abilities, anti-oxidative properties, and cell proliferations. It was observed that two polysaccharides with anti-oxidative effects by chelating metal ions reduced oxidative stress and further blocked the formation of reactive oxygen species syntheses. To detect whether there was a similar effect within the cellular mechanism, a flow cytometry was applied for sensing the oxidative level and it was found that both materials inhibited the endogenous oxidative stress, which was induced by phorbol-12-myristate-13-acetate (PMA). Both polysaccharides also stimulated the production of collagen to maintain skin tightness and a moisturizing effect. In summary, we developed two macromolecular polysaccharides with potential applications in dermal care.

Enriched Astaxanthin Extract from Haematococcus pluvialis Augments Growth Factor Secretions to Increase Cell Proliferation and Induces MMP1 Degradation to Enhance Collagen Production in Human Dermal Fibroblasts.

Among many antioxidants that are used for the repairing of oxidative stress induced skin damages, we identified the enriched astaxanthin extract (EAE) from Haematococcus pluvialis as a viable ingredient. EAE was extracted from the red microalgae through supercritical fluid carbon dioxide extraction. To compare the effectiveness, EAE wastreated on human dermal fibroblasts with other components, phorbol 12-myristate 13-acetate (PMA), and doxycycline. With sirius red staining and quantitative real-time polymerase chain reaction (qRT-PCR), we found that PMA decreased the collagen concentration and production while overall the addition of doxycycline and EAE increased the collagen concentration in a trial experiments. EAE increased collagen contents through inhibited MMP1 and MMP3 mRNA expression and induced TIMP1, the antagonists of MMPs protein, gene expression. As for when tested for various proteins through western blotting, it was seen that the addition of EAE increased the expression of certain proteins that promote cell proliferation. Testing those previous solutions using growth factor assay, it was noticeable that EAE had a positive impact on cell proliferation and vascular endothelial growth factor (VEGF) than doxycycline, indicating that it was a better alternative treatment for collagen production. To sum up, the data confirmed the possible applications as medical cosmetology agentsand food supplements.

Synergistic Effects of Linderanolide B Combined with Arbutin, PTU or Kojic Acid on Tyrosinase Inhibition.

Melanin uncontrollable accumulation is a serious social problem to not only women, but also men, and causes pigment over-expression disorders such as freckles, melasma or pigmented acne scars. The synergism is used widely in medication, and the effectiveness makes the drug applications more valuable. Within this experiment, three well-known compounds were chosen: kojic acid, 1-phenyl-2-thiourea (PTU) and arbutin, and they were combined individually with our substance linderanolide B, which is purified from Cinnamomum subavenium. Hence, deciphering the synergistic action of possible whitening agents was the goal of this study. The tyrosinase activity, melanin content, and the combination index (CI) values were observed in B16F10 cells, in addition, the consequences were detected by isobologram analysis. We discovered that certain melanin inhibitors showed synergistic properties when they were combined together to suppress tyrosinase activities. As a result, linderanolide B has a potential synergy on tyrosinase inhibition, and it can be used widely in cosmetic and medication industries.

Optimization of sodium loading on zeolite support for catalyzed transesterification of triolein with methanol.

Optimization of sodium loading on zeolite HY for catalyzed transesterification of triolein in excess methanol to biodiesel was studied. Zeolite HY catalyst was activated by loading sodium ions to their surface via an ion-exchange method. The effects of ion-exchange process parameters, including the temperature, the process time, the pH value, as well as concentrations and sources of Na(+) cations (NaOH, NaCl and Na2SO4), on the conversion yield of triolein to biodiesel were investigated. Most of these Na(+)-activated zeolite HY catalysts could really facilitate the catalyzed transesterification reaction of triolein to biodiesel at a lower temperature near 65°C. Consequently, a high conversion yield of triglycerides to biodiesel at 97.3% was obtained at 65°C. Moreover, the durability of zeolite catalysts was examined as well. Catalytic performance tests of these zeolite catalysts in transesterification did not show a significant decrease in catalysis at least for three batch cycles.