Evaluating the Antioxidants, Whitening and Antiaging Properties of Rice Protein Hydrolysates.

Plant-derived protein hydrolysates have potential applications in nutrition. Rice protein hydrolysates (RPHs), an excellent source of proteins, have attracted attention for the development of cosmeceuticals. However, few studies have reported the potential application of RPH in analysis, and this study examined their antioxidant activities and the inhibitory activities of skin aging enzymes. The results indicated that the total phenolic and flavonoid concentrations were 2.06 ± 0.13 mg gallic acid equivalent/g RPHs and 25.96 ± 0.52 µg quercetin equivalent/g RPHs, respectively. RPHs demonstrated dose-dependent activity for scavenging free radicals from 1,1-diphenyl-2-picrylhydrazyl [half-maximal inhibitory concentration (IC50) = 42.58 ± 2.1 mg/g RPHs] and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (IC50 = 2.11 ± 0.88 mg/g RPHs), dose-dependent reduction capacity (6.95 ± 1.40 mg vitamin C equivalent/g RPHs) and oxygen radical absorbance capacity (473 µmol Trolox equivalent/g RPHs). The concentrations of the RPH solution required to achieve 50% inhibition of hyaluronidase and tyrosinase activities were determined to be 8.91 and 107.6 mg/mL, respectively. This study demonstrated that RPHs have antioxidant, antihyaluronidase, and antityrosinase activities for future cosmetic applications.

Optimisation of the Production and Bleaching Process for a New Laccase from Madurella mycetomatis, Expressed in Pichia pastoris: from Secretion to Yielding Prominent.

Laccases are polyphenol oxidoreductases used in a number of industrial applications. Due to the increasing demand for these "green catalysis" enzymes, the identification and biochemical characterisation of their novel properties is essential. In our study, cloned Madurella mycetomatis laccase (mmlac) genes were heterologously expressed in the methylotrophic yeast host Pichia pastoris. The high yield of the active recombinant protein in P. pastoris demonstrates the efficiency of a reliably constructed plasmid to express the laccase gene. The optimal biochemical conditions for the successfully expressed MmLac enzyme were identified. Detailed structural properties of the recombinant laccase were determined, and its utility in decolourisation and textile bleaching applications was examined. MmLac demonstrates good activity in an acidic pH range (4.0-6.0); is stable in the presence of cationic metals, organic solvents and under high temperatures (50-60 °C); and is stable for long-term storage at - 20 °C and - 80 °C for up to eight weeks. The structural analysis revealed that the catalytic residues are partially similar to other laccases. MmLac resulted in an increase in whiteness, whilst demonstrating high efficiency and stability and requiring the input of fewer chemicals. The performance of this enzyme makes it worthy of investigation for use in textile biotechnology applications, as well as within environmental and food technologies.

Size-dependent whitening activity of enzyme-degraded fucoidan from Laminaria japonica.

Fucoidan from Laminaria japonica is a kind of sulfate polysaccharide with high molecular weight (MW) and broad bioactivities. This study was performed to investigate the relationship between MW and whitening activity of fucoidan and to exploit a novel functional ingredient for whitening cosmetics. High sulfate content fucoidan was enzymic degraded by Flavobacterium RC2-3 produced fucoidanase. Two hours were enough for the enzyme degradation to achieve degraded fucoidan with favorable tyrosinase inhibitory ability. The whitening activity of different MW fucoidan fractions were evaluated by their tyrosinase inhibitory ability, antioxidant activity and cellular melanogenesis inhibitory ability. Results showed that in the MW range above 5 kDa, the smaller MW of fucoidan were related to the better whitening activity. The fucoidan fraction with the MW between 5-10 kDa, presented the best tyrosinase inhibitory activity (62.0%), antioxidant activity (48.3%) and excellent anti-melanogenesis ability in B16 cells, which could be applied as the whitening factor in cosmetics development.

Semi-automated standardisation of melanin bleaching procedures of heavily pigmented melanocytic lesions for immunohistochemical analysis on an automated platform.

Background: The diagnosis of heavily pigmented melanocytic lesions is problematic. This is often compounded by lack of visibility of nuclear detail of tumour cells due to physical masking by melanin pigment. Similarly, there can be colour merging of chromogenic final reaction products with melanin, making an evidence of antigenic localisation problematic. There are a number of melanin bleaching techniques available for immunohistochemical assessments.Material and methods: All methods to date have involved the bleaching of melanin as a manually performed primary step before loading subsequently bleached slides onto automated immunohistochemical platforms. Here we define a semi-automated bleaching procedure that allows full integration on one of the most widely employed automated IHC staining platforms (Roche Ventana BenchMark Ultra). The bleaching protocol was defined on the BenchMark Ultra and involved the assessment of 24 histological cases of heavily pigmented malignant melanoma lesions (13 cutaneous and 11 metastatic) routinely fixed processed and paraffin wax embedded.Results: Completion of the bleaching was assessed on H&E preparations performed following the semi-automated bleaching step and employing the Roche Ventana BenchMark Ultra machine for 60 min at 42°C. Complete immunohistochemical staining was achieved on the automated platform within 5-6 h including the bleaching step. Results were consistent across all tissue evaluated.Discussion: This data provides evidence that the hydrogen peroxide bleaching procedure can be adapted for integration on one of the most widely employed automated IHC staining platforms and as a result, improve the efficiency and reproducibility of the technique.

L-765,314 Suppresses Melanin Synthesis by Regulating Tyrosinase Activity.

Although melanin production is a key self-defense mechanism against ultraviolet radiation (UVR)-induced skin damage, uneven or excessive deposition of melanin causes hyperpigmentary disorders. Currently available whitening agents are unsatisfactory because of issues with efficacy and safety. To develop more effective depigmenting agents, we performed high-throughput melanin content assay screening using the B16F10 melanoma cell line and identified L-765,314 as a drug that suppressed melanin production in cultured melanocytes in a dose-dependent manner as well as cAMP- or 12-O-tetradecanoylphorbol 13-acetate (TPA)-stimulated melanin production without cytotoxicity. Interestingly, melanogenic gene expression was not altered by L-765,314. Rather, diminished melanin production by L-765,314 appeared to be caused by downregulation of tyrosinase activity via inhibition of protein kinase C (PKC). Because L-765,314 did not show any adverse effect in melanocytes, altogether our data suggest that L-765,314 could be a potential therapeutic candidate for skin hyperpigmentary disorders and further discovery of selective inhibitors targeting PKC might be a promising strategy for the development of depigmenting agents to treat hyperpigmentary disorders.

Novel colour additive for bleach disinfectant wipes reduces corrosive damage on stainless steel.

Bleach disinfectant wipes are corrosive to hospital surfaces and equipment. This study measured the effect of two widely used bleach wipes, with and without Highlight® colour additive, on stainless steel to quantify the rate of corrosion and to determine the effect of Highlight® on reducing surface damage caused by bleach wipes. The two bleach wipes alone caused severe corrosion [>5 mils per year (mpy), where 1 mil = 0.001 inch], while the addition of Highlight® reduced the rate of corrosion significantly (<2 mpy) and prevented discolouration of the metal. These results indicate that Highlight® reduces the deleterious corrosive effects of bleach wipes, thus improving their viability for cleaning healthcare surfaces.

The presence of fecal test soil protects bacteria during cleaning/disinfection.

Reusable medical devices (RMDs) must be reprocessed between uses to render them safe for each use and each patient. Cleaning used devices removes organic and inorganic soil making them either safe for reuse or ready for disinfection/sterilization depending on the device. Although cleaning is an important step in a RMD's life cycle, it is not always a priority during device design. In addition, when performing cleaning validation, it is recommended that the manufacturer takes into consideration, what the most appropriate or worst case conditions are in terms of type of soil or the presence of bacteria. This study compared the ability of three different cleaning/disinfecting agents (water, alcohol, and bleach) to remove bacteria and fecal test soil from two different polymers: polypropylene and ultrahigh molecular weight polyethylene (UHMWPE) with two different roughness. There were some differences in the effects of the cleaning/disinfecting agents, the materials, and the roughness depending on the particular circumstances. However, the most consistent effect on the removal of bacteria was the presence of soil, which protected the bacteria from being removed. Conversely, the presence of bacteria played little role in the removal of soil. Although the interactions between material type and roughness, soil type, and bacteria are complicated, they should be taken into account during device design and reprocessing validation to create a device that is easy and safe to use. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1706-1710, 2019.

Melanin Bleaching With Warm Hydrogen Peroxide and Integrated Immunohistochemical Analysis: An Automated Platform.

OBJECTIVE: Diagnosing melanocytic lesions is among the most challenging problems in the practice of pathology. The difficulty of physically masking melanin pigment and the similarity of its color to commonly used chromogens often complicate examination of the cytomorphology and immunohistochemical staining results for tumor cells. Melanin bleach can be very helpful for histopathological diagnosis of heavily pigmented melanocytic lesions. Although various depigmentation methods have been reported, no standardized methods have been developed. This study developed a fully automated platform that incorporates hydrogen peroxide-based melanin depigmentation in an automated immunohistochemical analysis. METHODS AND MATERIALS: The utility of the method was tested in 1 cell block of malignant melanoma cells in pleural effusion, 10 ocular melanoma tissue samples, and 10 cutaneous melanoma tissue samples. Our results demonstrated that the proposed method, which can be performed in only 3 hours, effectively preserves cell cytomorphology and immunoreactivity. RESULTS: The method is particularly effective for removing melanin pigment to facilitate histopathological examination of cytomorphology and for obtaining an unmasked tissue section for immunohistochemical analysis.

Laccases, Manganese Peroxidases and Xylanases Used for the Bio-bleaching of Paper Pulp: An Environmental Friendly Approach.

BACKGROUND: The paper and pulp industry is a capital and resource-intensive industry that contributes to ecosystem toxicity and affects human beings. OBJECTIVE: The study aimed to appraise the potential of xylanases, laccases and manganese peroxidase for the bio-bleaching of paper pulp and to highlight the role of these enzymes as a promising substitute for chlorine-based chemical methods in the bleaching process. METHODS: The ligninolytic enzymes including xylanase, laccase and manganese peroxidase isolated from white rot fungi were used for pre-bleaching and bleaching of oven-dried wheat straw pulp. RESULTS: During the sequential enzymatic treatment of oven-dried pulp the brightness was improved and kappa number was reduced by 3.1% and 3.1 points respectively after xylanase treatment, 0.3% and 0.4 points after laccase treatment and 3% and 0.2 points after MnP treatment. During separate treatment of pulp samples with individual enzymes, brightness and kappa number improved by 8% and 3 points respectively after xylanase treatment, by 5% and 1.7 points after laccase treatment and 5% and 1.8 points after treatment with MnP. During subsequent treatment with 4% sodium hypochlorite, the brightness was further improved by 27.9 % for xylanase treated pulp and 29% for the laccase and MnP treated pulp. The xylanase was found most efficient in comparison to laccase and MnP in reduction of kappa number and improvement of brightness. CONCLUSION: These results clearly indicate the role of laccase, MnP and xylanase from white rot fungi as effective bio-bleaching agents. Therefore, these enzymes can facilitate the bleaching process without threat to environment.

Measuring antioxidant and prooxidant capacity using the Crocin Bleaching Assay (CBA).

The Crocin Bleaching Assay (CBA) appears in literature as an in vitro method for measuring antioxidant and prooxidant capacity of model dietary antioxidants, food formulations, pharmaceuticals, and biological samples. The assay is based on simple competitive reactions between a colored probe, crocin, and the test compounds/constituents for scavenging peroxyl radicals generated after thermolysis of a water-soluble azo-initiator. So far, several researchers in the fields of food chemistry, nutrition and clinical biochemistry have sporadically addressed critical views about advantages, limitations and potential field of CBA application. This chapter presents step-by-step critical aspects of CBA in order to assist standardization of its performance. Detailed procedures for calculation of two attributes of peroxyl radical scavenging reactions, the relative rate constant and "total antioxidant capacity", are also presented.

Study of melanin bleaching after immunohistochemistry of melanin-containing tissues.

Melanin may interfere with immunohistochemical staining. The goal of this study was to investigate the effects of trichloroisocyanuric acid (TCCA) bleaching, potassium permanganate bleaching, and potassium dichromate bleaching on melanin, tissue antigen, and 3,3'-diaminobenzidine (DAB) using melanin-containing and melanin-free tissue samples. Our results demonstrated that all 3 bleaching methods efficiently bleached melanin and partially destroyed tissue antigen. In addition, potassium permanganate bleaching and potassium dichromate bleaching clearly destroyed DAB, whereas TCCA bleaching had no significant effect on DAB. Therefore, neither potassium permanganate nor potassium dichromate is an ideal solution, whereas TCCA might be an ideal solution for melanin bleaching after the immunohistochemical staining of melanin-containing tissues. After immunostaining followed by TCCA bleaching, the melanin could be completely removed in all 120 malignant melanoma tissue sections. Compared with the control, the DAB intensity was clear, and the tissue structure and cellular nuclei were well maintained. It is worth noting that TCCA should be freshly prepared before each experiment, and used within 2 hours of its preparation. In addition, sections should not be incubated with TCCA for over 30 minutes.

Effective melanin depigmentation of human and murine ocular tissues: an improved method for paraffin and frozen sections.

PURPOSE: The removal of excessive melanin pigments that obscure ocular tissue morphology is important to address scientific questions and for differential diagnosis of ocular tumours based on histology. Thus, the goal of the present study was to establish an effective and fast melanin bleaching method for paraffin and frozen mouse and human ocular tissues. METHODS: Paraffin-embedded and frozen ocular specimens from mice and human donors were subjected to bleaching employing two methods. The first employed potassium permanganate (KMnO4) with oxalic acid, and the second 10% hydrogen peroxide (H2O2). To determine optimal bleaching conditions, depigmentation was carried out at various incubation times. The effect of diluents used for 10% H2O2 was assessed using phosphate-buffered saline (PBS), and deionized water. Three different slide types and two fixatives, which were ice-cold acetone with 80% methanol, and 4% paraformaldehyde (PFA) were used to determine the optimal conditions for better tissue adherence during bleaching. All tissues were stained in hematoxylin and eosin for histological evaluation. RESULTS: Optimal bleaching was achieved using warm 10% H2O2 diluted in PBS at 65°C for 120 minutes. Chromium-gelatin-coated slides prevented tissue detachment. Adherence of cryosections was also improved with post-fixation using 4% PFA and overnight air-drying at RT after cryosectioning. Tissue morphology was preserved under these conditions. Conversely, tissues bleached in KMnO4/oxalic acid demonstrated poor depigmentation with extensive tissue damage. CONCLUSIONS: Warm dilute H2O2 at 65°C for 120 minutes rapidly and effectively bleached both cryo- and paraffin sections of murine and human ocular tissues.

Influence of exposure time to saliva and antioxidant treatment on bond strength to enamel after tooth bleaching: an in situ study

Objectives: This study evaluated the influence of different exposure times to saliva in situ in comparison with an antioxidant treatment on composite resin bond strength to human enamel restored after tooth bleaching. Material and Methods: Forty human teeth specimens measuring 5x5 mm were prepared and randomly allocated into 5 groups with 8 specimens each: Gct (control group, restored on unbleached enamel); Gbl (restored immediately after bleaching); Gsa (bleached, treated with 10% sodium ascorbate gel for 60 min and restored); G7d (bleached, exposed to saliva in situ for 7 days and restored); and G14d (bleached, exposed to saliva in situ for 14 days and restored). Restored samples were cut into 0.8 mm2 sticks that were tested in microtensile. Specimens were microscopically analyzed and failure modes were classified as adhesive, cohesive, or mixed. Pretest and cohesive failures were not considered in the statistical analysis, which was performed with one-way ANOVA and Tukey's post-hoc test (α=0.05), with the dental specimen considered as the experimental unit. Results: Mean bond strength results found for Gbl in comparison with Gct indicated that bleaching significantly reduced enamel adhesiveness (P<0.01). However, no statistically significant differences were found between Gct, Gsa and G7d (P>0.05). Bond strength found for G14d was significantly higher than for Gsa (P<0.01). Fractures modes were predominantly of a mixed type. Conclusions: Bonding strength to bleached enamel was immediately restored with the application of sodium ascorbate and exposure to human saliva in situ for at least 7 days. Best results were obtained with exposure to human saliva in situ for 14 days. Treatment with sodium ascorbate gel for 60 min may be recommended in cases patients cannot wait for at least 7 days for adhesive techniques to be performed. .

Chemical cleaning agents and bonding to glass-fiber posts

The influence of chemical cleaning agents on the bond strength between resin cement and glass-fiber posts was investigated. The treatments included 10% hydrofluoric acid, 35% phosphoric acid, 50% hydrogen peroxide, acetone, dichloromethane, ethanol, isopropanol, and tetrahydrofuran. Flat glass-fiber epoxy substrates were exposed to the cleaners for 60 s. Resin cement cylinders were formed on the surfaces and tested in shear. All treatments provided increased bond strength compared to untreated control specimens. All failures were interfacial. Although all agents improved the bond strength, dichloromethane and isopropanol were particularly effective.

The impact of iron on the bleaching efficacy of hydrogen peroxide in liquid whey systems.

UNLABELLED: Whey is a value-added product that is utilized in many food and beverage applications for its nutritional and functional properties. Whey and whey products are generally utilized in dried ingredient applications. One of the primary sources of whey is from colored Cheddar cheese manufacture that contains the pigment annatto resulting in a characteristic yellow colored Cheddar cheese. The colorant is also present in the liquid cheese whey and must be bleached so that it can be used in ingredient applications without imparting a color. Hydrogen peroxide and benzoyl peroxide are 2 commercially approved chemical bleaching agents for liquid whey. Concerns regarding bleaching efficacy, off-flavor development, and functionality changes have been previously reported for whey bleached with hydrogen peroxide and benzoyl peroxide. It is very important for the dairy industry to understand how bleaching can impact flavor and functionality of dried ingredients. Currently, the precise mechanisms of off-flavor development and functionality changes are not entirely understood. Iron reactions in a bleached liquid whey system may play a key role. Reactions between iron and hydrogen peroxide have been widely studied since the reaction between these 2 relatively stable species can cause great destruction in biological and chemical systems. The actual mechanism of the reaction of iron with hydrogen peroxide has been a controversy in the chemistry and biological community. The precise mechanism for a given reaction can vary greatly based upon the concentration of reactants, temperature, pH, and addition of biological material. In this review, some hypotheses for the mechanisms of iron reactions that may occur in fluid whey that may impact bleaching efficacy, off-flavor development, and changes in functionality are presented. PRACTICAL APPLICATION: Cheese whey is bleached to remove residual carotenoid cheese colorant. Concerns regarding bleaching efficacy, off-flavor development, and functionality changes have been reported for whey proteins bleached with hydrogen peroxide and benzoyl peroxide. It is very important for the dairy industry to understand how whey bleaching can impact flavor and functionality of dried ingredients. Proposed mechanisms of off-flavor development and functionality changes are discussed in this hypothesis paper.

Biological activity of bleached kraft pulp mill effluents before and after activated sludge and ozone treatments.

Eucalyptus bleached kraft pulp production, an important sector of the Brazilian national economy, is responsible for generating large volume, high pollutant load effluents, containing a considerable fraction of recalcitrant organic matter. The objectives of this study were to quantify the biological activity of the effluent from a eucalyptus bleached kraft pulp mill, characterize the nature of compounds responsible for biological activity and assess the effect of ozone treatment on its removal. Primary and secondary effluents were collected bimonthly over the course of one year at a Brazilian bleached eucalypt kraft pulp mill and their pollutant loads (biochemical oxygen demand (BOD), chemical oxygen demand (COD), total organic carbon (TOC), adsorbable organic halogen (AOX), lignin, extractives) and biological activity (acute and chronic toxicity and estrogenic activity) quantified. The effluent studied did not present acute toxicity to Daphnia, but presented the chronic toxicity effects of algal growth inhibition and reduced survival and reproduction in Ceriodaphnia, as well as estrogenic activity. Chronic toxicity and estrogenic activity were reduced but not eliminated during activated sludge biological treatment. The toxicity identification evaluation revealed that lipophilic organic compounds (such as residual lignin, extractives and their byproducts) were responsible for the toxicity and estrogenic activity. Ozone treatment (50 mg/L O(3)) of the secondary effluent eliminated the chronic toxicity and significantly reduced estrogen activity.

Isolation, fractionation and characterization of melanin-like pigments from chestnut (Castanea mollissima) shells.

UNLABELLED: Melanins are known as versatile biopolymers, but the utilizations are restricted by their poor solubilities. Therefore, well soluble ones or their analogs are much desired. In this article, a new procedure was developed for fractionation of the pigments isolated from chestnut (Castanea mollissima) shells, and 3 fractions (Fr. 1, Fr. 2, and Fr. 3) were obtained. The solubilities of all the fractions in waters of different pH and in common organic solvents were studied. The physicochemical properties of the fractions were characterized for the first time on the basis of combined chemical analyses and spectroscopic methods including ultraviolet-visible (UV-Vis), Fourier transform infrared (FT-IR), electron spin resonance (ESR), and solid-state ¹³C nuclear magnetic resonance (¹³C-NMR). All the fractions could be bleached by NaOCl and H2O2 and give a positive reaction for polyphenols, which are usually used as typical tests for allomelanins. Their UV-Vis, FT-IR, and ESR spectra resembled those of synthetic and some natural melanins. Elemental data and quantitative analyses of ¹³C-NMR spectra revealed that pigment-bound proteins and polysaccharides were the most abundant in Fr. 1, while Fr. 2 was presented with the highest aromaticity. PRACTICAL APPLICATION: We provided a new, simple, and inexpensive method to fractionate the melanin-like pigments from chestnut shells. This technique can be used to produce natural melanin-like food colorants with different solubilities from chestnut shells.

Diffuse reflectance spectroscopy of fibrous proteins.

UV-visible diffuse reflectance (DR) spectra of the fibrous proteins wool and feather keratin, silk fibroin and bovine skin collagen are presented. Natural wool contains much higher levels of visible chromophores across the whole visible range (700-400 nm) than the other proteins and only those above 450 nm are effectively removed by bleaching. Both oxidative and reductive bleaching are inefficient for removing yellow chromophores (450-400 nm absorbers) from wool. The DR spectra of the four UV-absorbing amino acids tryptophan, tyrosine, cystine and phenylalanine were recorded as finely ground powders. In contrast to their UV-visible spectra in aqueous solution where tryptophan and tyrosine are the major UV absorbing species, surprisingly the disulphide chromophore of solid cystine has the strongest UV absorbance measured using the DR remission function F(R)(∞). The DR spectra of unpigmented feather and wool keratin appear to be dominated by cystine absorption near 290 nm, whereas silk fibroin appears similar to tyrosine. Because cystine has a flat reflectance spectrum in the visible region from 700 to 400 nm and the powder therefore appears white, cystine absorption does not contribute to the cream colour of wool despite the high concentration of cystine residues near the cuticle surface. The disulphide absorption of solid L: -cystine in the DR spectrum at 290 nm is significantly red shifted by ~40 nm relative to its wavelength in solution, whereas homocystine and lipoic acid showed smaller red shifts of 20 nm. The large red shift observed for cystine and the large difference in intensity of absorption in its UV-visible and DR spectra may be due to differences in the dihedral angle between the crystalline solid and the solvated molecules in solution.

Effect of high-concentrated bleaching agents on the bond strength at dentin/resin interface and flexural strength of dentin

This study evaluated the effect of bleaching agents on bond strength at the dentin/resin interface and the flexural strength of dentin. Forty maxillary canines were selected for the study. In the shear strength test, 40 slabs of intracoronary dentin (5 x 5 mm) obtained from buccal surfaces of the crowns were included in acrylic resin. In the flexural strength test, 40 dentin bars (8 x 2 x 2 mm) were obtained from the roots. The 40 hemi-sections of the lingual surface were prepared for scanning electron microscopy (SEM). The specimens were divided into 4 groups according to the bleaching protocol (n=10): Unbleached (control), Sodium perborate + 20 percent hydrogen peroxide (SP + 20 percent HP), 37 percent carbamide peroxide (37 percent CP) and 38 percent hydrogen peroxide (38 percent HP). After 7 days, the bond strength specimens were restored and tested. Dentin bars were bleached and subjected to a three-point bending test. Data (MPa) were analyzed by ANOVA and Tukey's test (α=0.05). In the shear test, the control group was superior (p<0.05) to the bleached groups, which, in turn, were statistically similar (p>0.05). In the flexural strength test, the control group also had the highest values and differed significantly from the other groups (p<0.05). SEM revealed smear layer in all groups, with fissures in the bleached specimens. SP + 20 percent HP and 38 percent HP showed discontinuous interfaces with few tags. In conclusion, bond strength of restorative material to dentin and flexural strength of dentin were reduced after the use of high-concentration bleaching agents.

Este estudo avaliou o efeito de agentes clareadores na resistência de união da interface dentina/resina e resistência à flexão da dentina. Quarenta caninos superiores foram selecionados para o estudo. No teste de cisalhamento, 40 fragmentos de dentina intracoronária (5 x 5 mm) obtidos a partir de superfícies vestibulares das coroas foram incluídos em resina acrílica. No teste de flexão, 40 barras de dentina (8 x 2 x 2 mm) foram obtidas a partir das raízes. As 40 hemi-seções da superfície lingual foram preparadas para microscopia eletrônica de varredura (MEV). Os espécimes foram divididos em 4 grupos de acordo com o protocolo de clareamento (n=10): Não clareados (controle), perborato de sódio + peróxido de hidrogênio 20 por cento (PS + PH 20 por cento), peróxido de carbamida 37 por cento (PC37 por cento) e peróxido de hidrogênio 38 por cento (PH 38 por cento). Após 7 dias, as amostras destinadas à resistência de união foram restauradas e submetidas ao teste. As barras de dentina foram clareadas e submetidas ao teste de flexão de 3 pontos. Os dados (MPa) foram analisados por ANOVA e teste de Tukey (α=0,05). No teste de cisalhamento, o controle foi superior (p<0,05) aos grupos clareados, que foram semelhantes entre si (p>0,05). Na resistência à flexão, o grupo controle também exibiu os maiores valores, diferente dos demais (p<0,05). SEM revelou camada de smear em todos os grupos, com fissuras nos espécimes clareados. PS + 20 por cento PH e PH 38 por cento apresentaram interfaces de descontínuas com poucos tags. A resistência de união do material restaurador à dentina e a resistência à flexão da dentina foram reduzidas após o uso de agentes clareadores de alta concentração.

Partially purified components of Nardostachys chinensis suppress melanin synthesis through ERK and Akt signaling pathway with cAMP down-regulation in B16F10 cells.

UNLABELLED: Ethnopharmacological relevance Nardostachys chinensis has been used in folk medicine to treat melasma and lentigines in Korea. We investigated the inhibitory activities of Nardostachys chinensis in melanogenesis and its related signaling pathway. MATERIALS AND METHODS: Bioassay-guided fractionation of Nardostachys chinensis using solvent partitioning and purification with octadecylsilane open-column chromatography resulted in partial purification. The active 20% methanol chromatographic fraction from the ethyl acetate layer (PPNC) was used to investigate melanogenesis by melanin synthesis, tyrosinase activity assay, cAMP assay, Western blot and flow cytometric analyses in B16F10 mouse melanoma cells. RESULTS: PPNC markedly inhibits melanin synthesis and tyrosinase activity in a concentration-dependent manner. We also found that PPNC decreases microphthalmia-associated transcription factor (MITF), tyrosinase, tyrosinase-related protein (TRP)-1, and dopachrome tautomerase (Dct) protein expressions and MITF and tyrosinase mRNA levels. Moreover, PPNC reduces intracellular cAMP levels and activates mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) and phosphatidylinositol 3-kinase (PI3K)/Akt expression in B16F10 cells. The specific MEK/ERK inhibitor PD98059 and PI3K/Akt inhibitor LY294002, block the PPNC-induced hypopigmentation effect, and abrogate the PPNC-suppressed expression of melanogenic proteins such as MITF, tyrosinase, TRP-1, and Dct. Using flow cytometry, we elucidated whether PPNC directly induces ERK phosphorylation at the level of an intact single cell. PPNC shows marked expression of phosphorylated ERK in live B16F10 cells and abrogates PPNC-induced phosphorylated ERK by PD98059 treatment. CONCLUSIONS: PPNC stimulates MEK/ERK phosphorylation and PI3K/Akt signaling with suppressing cAMP levels and subsequently stimulating MITF and TRPs down-regulation, resulting in melanin synthesis suppression.