Silencing Stem Cell Factor Gene in Fibroblasts to Regulate Paracrine Factor Productions and Enhance c-Kit Expression in Melanocytes on Melanogenesis.

Melanogenesis is a complex physiological mechanism involving various paracrine factors. Skin cells such as keratinocytes, fibroblasts, and melanocytes communicate with one another through secreted regulators, thereby regulating the melanocytes' bio-functions. The stem cell factor (SCF) is a paracrine factor produced by fibroblasts, and its receptor, c-kit, is expressed on melanocytes. Binding of SCF to c-kit activates autophosphorylation and tyrosine kinase to switch on its signal transmission. SCF inhibition does not suppress fibroblast proliferation in MTT assay, and SCF silencing induced mRNA expressions of paracrine factor genes, HGF, NRG-1, and CRH in qPCR results. Following UVB stimulation, gene expressions of HGF, NRG, and CRH were higher than homeostasis; in particular, HGF exhibited the highest correlation with SCF variations. We detected fibroblasts regulated SCF in an autocrine-dependent manner, and the conditioned medium obtained from fibroblast culture was applied to treat melanocytes. Melanogenesis-related genes, tyrosinase and pmel17, were upregulated under conditioned mediums with SCF silencing and exposed to UVB treatments. Melanin quantities in the melanocytes had clearly increased in the pigment content assay. In conclusion, SCF silencing causes variations in both fibroblast paracrine factors and melanocyte melanogenesis, and the differences in gene expressions were observed following UVB exposure.

Composite grafting with pulp adipofascial advancement flaps for treating non-replantable fingertip amputations.

BACKGROUND: Non-replantable fingertip amputation is still a clinical challenge. We performed modified composite grafting with pulp adipofascial advancement flap for Hirase IIA fingertip amputations. Results from a series of patients are presented and achieved better outcome than traditional composite grafting. PATIENTS AND METHODS: From September 2012 to April 2014, fourteen patients with sixteen digits were included in our study. Mean age of patients was 43.9 years (20-71 years). All of our patients underwent this procedure under digital block anesthesia. We performed pulp adipofascial advancement flap for better soft tissue coverage of bone exposure stump first. The amputated parts were defatted, trimming, and reattached as composite graft. Age and gender of patients, injured finger, Hirase classification, mechanism of trauma, overall graft survival area, two-point discrimination (2PD) (mm) at six-month, length of shortening of digit, The average disabilities of the arm, shoulder, and hand (DASH) score and subjective self-evaluation questionnaire at 6 month were recorded. RESULTS: Average graft survival area was 89% (75-100%). Average length of shortening was 2.2 mm (1.8-3.5 mm). 2PD at six-month after surgery was 6.3 mm in average (5-8 mm). Average DASH score at 6 month was 1.45 (0.83-2.5). The self-evaluated aesthetic results showed twelve patients (85.7%) were very satisfied, and no patient was completely unsatisfied. CONCLUSIONS: In Hirase zone IIA traumatic fingertip amputation where replantation is difficult, our modified technique of composite grafting with pulp adipofascial advancement flap provided an alternative choice with high successful rate, acceptable functional and aesthetic outcomes. © 2016 Wiley Periodicals, Inc. Microsurgery, 2016. © 2015 Wiley Periodicals, Inc. Microsurgery 36:651-657, 2016.

Extrusion puffing pretreated cereals for rapid production of high-maltose syrup.

In this study, cereals with high starch content, including brown rice, corn, and buckwheat were pretreated by extrusion. The physicochemical properties of extruded-puffed cereals obtained from different extrusion conditions were analyzed herein. The puffed extrudates exhibited lower bulk density, higher water solubility and gelatinization as compared to untreated cereals. The FTIR-ATR results confirmed a decrease in the crystalline structure of extruded-puffed cereals. A higher Vmax/Km value was observed in the enzymatic saccharification of puffed extrudates that significantly improved hydrolysis rate and yield. Finally, the high-maltose syrup was produced via the enzymatic hydrolysis of extruded-puffed cereals at high substrate concentrations (20 %). After hydrolysis for 180 min at an enzyme substrate ratio (E/S ratio) of 0.2, the syrup with dextrose equivalent (DE) value of 63, 62, and 61 were obtained from extruded-puffed brown rice, corn, and buckwheat, respectively. Our results showed the potential of using extruded-puffed cereals for producing high-maltose syrup.

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.

Extraction and characterization of phenolic compounds with antioxidant and antimicrobial activities from pickled radish.

The pickled radish can be kept at room temperature for years without spoilage. 2,6-dihydroxyacetophenone (DHAP), 4-hydroxybenzaldehyde (HBA), and 4-hydroxyphenethyl alcohol (4-HPEA) were first found from the pickled radish. The structures of three phenolic compounds were elucidated by analysis of their nuclear magnetic resonance and high-resolution electro-spray ionization mass spectrometry data. All these phenolic compounds showed good free radical scavenging capacity except HBA. Both DHAP and 4-HPEA also showed high ferric reducing ability. DHAP showed good antimicrobial activity against Escherichia coli, Bacillus subtilis, and Canidia albicans. HBA demonstrated antimicrobial activity against E. coli and C. albicans but not B. subtilis. Based on the results of MTT assay, these compounds did not show cytotoxicity to LO2 cell line. All results indicated the pickled radish had antioxidant and antimicrobial phenolic compounds. To the best of our knowledge, this report is the first to answer partially the question of why pickled foods can be kept at room temperature for years without spoilage based on the evidence of three phenolic compounds.

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.

A novel polysaccharide from Malus halliana Koehne with coagulant activity.

A novel polysaccharide in Malus halliana Koehne, named MHP-W, was isolated and purified by DEAE-52 cellulose and Sephadex G-100 columns. Structural features were identified by high performance size-exclusion chromatography (HPSEC), fourier transform infrared (FT-IR) spectrometer, gas chromatography (GC) and (1D & 2D) NMR Spectroscopy. Structural characterization showed that the molecular weight of MHP-W was 353 kDa composed of arabinose, xylose, mannose, glucose and galactose in a molar ratio of 2.59: 0.15: 0.23: 0.25: 9.70. The existence of ß-glycosidic bond between the sugar units was confirmed by FT-IR and NMR spectroscopy. The effects of MHP-W on active part thrombin time (APTT), protothrombin time (PT), thrombin time (TT), and fibrinogen (FIB) were screened by a cell-based coagulation activity model. MHP-W could significantly shorten TT (p < 0.001) and increase FIB (p < 0.05) as compared with the control group. The results showed that MHP-W promoted bloodclotting through endogenous and exogenous coagulation pathways as well as increasing fibrinogen content, which indicated that MHP-W had procoagulant activities in vitro.

Investigations of kanuka and manuka essential oils for in vitro treatment of disease and cellular inflammation caused by infectious microorganisms.

BACKGROUND: Diseases caused by infectious and inflammatory microorganisms are among the most common and most severe nosocomial diseases worldwide. Therefore, developing effective agents for treating these illnesses is critical. In this study, essential oils from two tea tree species, kanuka (Kunzea ericoides) and manuka (Leptospermum scoparium), were evaluated for use in treating diseases and inflammation caused by microorganism infection. METHODS: Isolates of clinically common bacteria and fungi were obtained from American Type Culture Collection and from Kaohsiung Veterans General Hospital. Minimum inhibitory concentrations for Trichosporon mucoides, Malassezia furfur, Candida albicans, and Candida tropicalis were determined by the broth microdilution method with Sabouraud dextrose broth. The antibacterial susceptibility of Staphylococcus aureus, Streptococcus sobrinus, Streptococcus mutans, and Escherichia coli were determined by the broth microdilution method. A human acute monocytic leukemia cell line (THP-1) was cultured to test the effects of the essential oils on the release of the two inflammatory cytokines, tumor necrosis factor-α and interleukin-4. RESULTS: Multiple analyses of microorganism growth confirmed that both essential oils significantly inhibited four fungi and the four bacteria. The potent fungicidal properties of the oils were confirmed by minimum inhibitory concentrations ranging from 0.78% to 3.13%. The oils also showed excellent bactericidal qualities with 100% inhibition of the examined bacteria. In THP-1 cells, both oils lowered tumor necrosis factor-α released after lipopolysaccharide stimulation. Finally, the antimicrobial and anti-inflammatory effects of the oils were obtained without adversely affecting the immune system. CONCLUSION: These results indicate that the potent antimicroorganism and anti-inflammation properties of kanuka and manuka essential oils make them strong candidates for use in treating infections and immune-related disease. The data confirm the potential use of kanuka and manuka extracts as pharmaceutical antibiotics, medical cosmetology agents, and food supplements.

An Acetamide Derivative as a Camptothecin Sensitizer for Human Non-Small-Cell Lung Cancer Cells through Increased Oxidative Stress and JNK Activation.

In recent years, combination chemotherapy is a primary strategy for treating lung cancer; however, the issues of antagonism and side effects still limit its applications. The development of chemosensitizer aims to sensitize chemoresistant cancer cells to anticancer drugs and therefore improve the efficacy of chemotherapy. In this study, we examined whether N-[2-(morpholin-4-yl)phenyl]-2-{8-oxatricyclo[7.4.0.0,2,7]trideca-1(9),2(7),3,5,10,12-hexaen-4-yloxy}acetamide (NPOA), an acetamide derivative, sensitizes human non-small-cell lung cancer (NSCLC) H1299 cells towards camptothecin- (CPT-) induced apoptosis effects. Our results demonstrate that the combination of CPT and NPOA enhances anti-lung-cancer effect. The cytometer-based Annexin V/propidium iodide (PI) staining showed that CPT and NPOA cotreatment causes an increased population of apoptotic cells compared to CPT treatment alone. Moreover, Western blotting assay showed an enhancement of Bax expression and caspase cascade leading to cell death of H1299 cells. Besides, CPT and NPOA cotreatment-mediated disruption of mitochondrial membrane potential (MMP) in H1299 cells may function through increasing the activation of the stressed-associated c-Jun N-terminal kinase (JNK). These results showed that NPOA treatment sensitizes H1299 cells towards CPT-induced accumulation of cell cycle S phase and mitochondrial-mediated apoptosis through regulating endogenous ROS and JNK activation. Accordingly, NPOA could be a candidate chemosensitizer of CPT derivative agents such as irinotecan or topotecan in the future.

Polygonum cuspidatum extracts as bioactive antioxidaion, anti-tyrosinase, immune stimulation and anticancer agents.

In our study, it was applied for the technology of supercritical fluid carbon dioxide extraction to achieve biological constitutes from a Taiwan native plant, Polygonum cuspidatum. We developed bioactive effects of P. cuspidatum extracts via multiple examinations that established bio-purposes at a range of dosage ranges. The research of P. cuspidatum extracts indicated that they possessed anti-oxidative properties on radical-scavenging abilities, reducing activities and metal chelating powers in dose-dependant manners. The extracts also had minor in vitro mushroom tyrosinase suppression and decreased cellular tyrosinase activities and melanin production in B16-F10 cells. Immunologically, P. cuspidatum extracts enhanced the release of tumor necrosis factor α (TNF-α) induced by THP-1 macrophage cell line. In addition, the cell proliferation showed anti-proliferation in dose-dependent manner on human skin melanoma cells, A375 and A375.S2, of the extracts suggesting biological constitutes employed the anti-cancer possessions. This is the first statement presenting bioactivities on P. cuspidatum extracts including anti-oxidation, immune stimulation, anti-tyrosinase and anti-melanoma as far as we know.

The Effect in Topical Use of Lycogen(TM) via Sonophoresis for Anti-aging on Facial Skin.

BACKGROUND: Anti-aging skin care is a growing popular topic in cosmetic and aesthetic fields, and skin care rather then makeup tips draw more attention nowadays. The phenomenon of skin aging includes thinning of skin losses of elasticity and moisture, pigmented spot formation, and wrinkle development. Along with growth in age, the decreased rates of epithelium renewal and cellular recovery as well as the reduced contents of elastin, collagen, and glycosaminoglycans all contribute to creases or folds of skin. Available strategies for wrinkle treatments include topical use of skin care products with anti-aging contents, dermabrasion, laser, Botox injection, fillers injection, and facelift. Though all of these above options can provide different degrees of improvement in facial wrinkles, the cost-effect, pain of intervention therapy, and necessity of repetitive treatment may impact on choices made. Topical use of anti-aging skin products is the most convenient and cheap way to achieve skin anti-aging effect. Lycogen(TM) is an antioxidant, which can prevent the downregulation of pro-collagen I, intracellular accumulation of malondialdehyde (MDA) and achieve the aim of skin rejuvenation. METHODS: Twenty-six female patients were included in our study with ages between 30 and 45. They were randomly assigned to two groups: the vehicle control group and the experimental group. Patients in the control group applied a skin care product without Lycogen(TM)to the face via sonophoresis after facial cleanser use in the morning and at night. The experimental group applied a Lycogen(TM) -containing skin care product via sonophoresis in the same time schedule. We evaluated results, including pigmented spots, wrinkles, texture, pores, and red area by VISIA on weeks 0, 1, 2, 4, 6, 8, and 10 respectively. RESULTS: In the aspect of pigmented spots, the experimental group showed significant difference in comparison with the vehicle control group on weeks 2, 6, 8, and 10. For wrinkles, the experimental group had better results on weeks 1, 2, 4, 8, and 10. Measured by texture, the experimental group had better results on weeks 1, 2, 4, 6, 8, and 10. Determined by pores, the experimental group had better results on weeks 2, 4, 6, 8, and 10. Concerning red areas, the experimental group had better results on weeks 6, 8, and 10. (p < 0.05). CONCLUSION: In our study, we applied a Lycogen(TM)- containing product by sonophoresis as the experimental group in comparison with a skin care product without LycogenTM. VISIA (Canfield Imaging Systems, Fairfield, NJ) was used to evaluate facial skin in aspects of pigmented spots, wrinkles, texture, pores, and red area. Overall, Lycogen(TM) had proven effectiveness on anti-oxidation as patients who used the Lycogen TM -containing product had better outcomes.

Fat Grafting in Burn Scar Alleviates Neuropathic Pain via Anti-Inflammation Effect in Scar and Spinal Cord.

Burn-induced neuropathic pain is complex, and fat grafting has reportedly improved neuropathic pain. However, the mechanism of fat grafting in improving neuropathic pain is unclear. Previous investigations have found that neuroinflammation causes neuropathic pain, and anti-inflammatory targeting may provide potential therapeutic opportunities in neuropathic pain. We hypothesized that fat grafting in burn scars improves the neuropathic pain through anti-inflammation. Burn-induced scar pain was confirmed using a mechanical response test 4 weeks after burn injuries, and autologous fat grafting in the scar area was performed simultaneously. After 4 weeks, the animals were sacrificed, and specimens were collected for the inflammation test, including COX-2, iNOS, and nNOS in the injured skin and spinal cord dorsal horns through immunohistochemistry and Western assays. Furthermore, pro-inflammatory cytokines (IL-1 ß and TNF-α) in the spinal cord were collected. Double immunofluorescent staining images for measuring p-IκB, p-NFκB, p-JNK, and TUNEL as well as Western blots of AKT, Bax/Bcl-2 for the inflammatory process, and apoptosis were analyzed. Fat grafting significantly reduced COX2, nNOS, and iNOS in the skin and spinal cord dorsal horns, as well as IL-1ß and TNF-α, compared with the burn group. Moreover, regarding the anti-inflammatory effect, the apoptosis cells in the spinal cord significantly decreased after the fat grafting in the burn injury group. Fat grafting was effective in treating burn-induced neuropathic pain through the alleviation of neuroinflammation and ameliorated spinal neuronal apoptosis.

An iridium(iii)-based irreversible protein-protein interaction inhibitor of BRD4 as a potent anticancer agent.

Bromodomain-containing protein 4 (BRD4) has recently emerged as an attractive epigenetic target for anticancer therapy. In this study, an iridium(iii) complex is reported as the first metal-based, irreversible inhibitor of BRD4. Complex 1a is able to antagonize the BRD4-acetylated histone protein-protein interaction (PPI) in vitro, and to bind BRD4 and down-regulate c-myc oncogenic expression in cellulo. Chromatin immunoprecipitation (ChIP) analysis revealed that 1a could modulate the interaction between BRD4 and chromatin in melanoma cells, particular at the MYC promoter. Finally, the complex showed potent activity against melanoma xenografts in an in vivo mouse model. To our knowledge, this is the first report of a Group 9 metal complex inhibiting the PPI of a member of the bromodomain and extraterminal domain (BET) family. We envision that complex 1a may serve as a useful scaffold for the development of more potent epigenetic agents against cancers such as melanoma.