The SH-SY5Y human neuroblastoma cell line, a relevant in vitro cell model for investigating neurotoxicology in human: Focus on organic pollutants.

Investigation of the toxicity triggered by chemicals on the human brain has traditionally relied on approaches using rodent in vivo models and in vitro cell models including primary neuronal cultures and cell lines from rodents. The issues of species differences between humans and rodents, the animal ethical concerns and the time and cost required for neurotoxicity studies on in vivo animal models, do limit the use of animal-based models in neurotoxicology. In this context, human cell models appear relevant in elucidating cellular and molecular impacts of neurotoxicants and facilitating prioritization of in vivo testing. The SH-SY5Y human neuroblastoma cell line (ATCC® CRL-2266™) is one of the most used cell lines in neurosciences, either undifferentiated or differentiated into neuron-like cells. This review presents the characteristics of the SH-SY5Y cell line and proposes the results of a systematic review of literature on the use of this in vitro cell model for neurotoxicity research by focusing on organic environmental pollutants including pesticides, 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD), flame retardants, PFASs, parabens, bisphenols, phthalates, and PAHs. Organic environmental pollutants are widely present in the environment and increasingly known to cause clinical neurotoxic effects during fetal & child development and adulthood. Their effects on cultured SH-SY5Y cells include autophagy, cell death (apoptosis, pyroptosis, necroptosis, or necrosis), increased oxidative stress, mitochondrial dysfunction, disruption of neurotransmitter homeostasis, and alteration of neuritic length. Finally, the inherent advantages and limitations of the SH-SY5Y cell model are discussed in the context of chemical testing.

Óleo essencial de Pataqueira (Conobea scoparioides Cham. & Schltdl.) como conservante natural em Cosméticos / Pataqueira essential oil (Conobea scoparioides Cham. & Schltdl.) as a natural preservative in cosmetics

Um dos principais grupos de conservantes utilizados na maioria dos cosméticos são os parabenos que em muitos estudos demonstraram que podem provocar reações alérgicas como dermatite de contato, entre outras sensibilizações cutâneas. A fim de minimizar esses problemas, a indústria está produzindo cosméticos livres de conservantes ou de origem natural e em associações aos sintéticos. Dentre os conservantes naturais utilizados, podemos citar os óleos essenciais como uma alternativa viável. Diante deste contexto o presente trabalho visa avaliar experimentalmente o potencial antimicrobiano do óleo essencial de Conobea scoparioides Cham. & Schltdl., conhecida popularmente como pataqueira, o efeito de sua associação com parabenos e de sua eficácia como conservante em bases cosméticas. A composição do óleo essencial foi avaliada, indicando que este é composto em sua maior parte por terpenos, tendo éter metílico do timol (39,2%), timol (33,8 %) e α-felandreno (15,9%) como compostos majoritários. A atividade antimicrobiana do óleo essencial e do timol foi acessada através da concentração inibitória mínima (CIM), cujos resultados em µg/mL para o óleo essencial e o timol foram respectivamente: Staphylococcus aureus 650,70 e 284,90, Escherichia coli 721,53 e 271,20, Pseudomonas aeruginosa 1748,00 e > 2.000, Burkholderia cepacia 833,03 e 1.077,70, Candida albicans 521,43 e 172,61 e Aspergillus brasiliensis 300 e 400. O efeito sinérgico da associação do óleo essencial com os parabenos foi realizado através de um delineamento experimental centroide simplex para uma mistura de metilparabeno, propilparabeno e óleo essencial frente aos mesmos micro-organismos utilizados na determinação da atividade antimicrobiana. As concentrações ideais obtidas pela análise estatística para cada componente em µg/mL foram: 1120 para o metilparabeno, 350 para o propilparabeno e 675 para o óleo essencial. O teste de eficácia do sistema conservante em formulação cosmética foi efetuado empregando as concentrações ideais e mais duas concentrações superiores e uma abaixo do ideal. Para todas as cepas microbianas desafiadas o resultado do teste foi de redução total da carga microbiana inoculada nos sete dias de ensaio e nenhum aumento até o vigésimo oitavo dia o que demonstra a eficácia da associação do óleo essencial com os conservantes sintéticos. O óleo essencial de C. scoparioides apresentou um potencial antimicrobiano importante tanto sozinho como em associação com conservantes sintéticos. Estes resultados sugerem que esse óleo pode ser usado para compor um sistema conservante para formulações cosméticas contendo uma menor quantidade de sintéticos

One of the main groups of preservatives used in most cosmetics are parabens, that many studies have shown that they can cause allergic reactions such as contact dermatitis, among other skin sensitizations. To minimize these problems, the industry is producing cosmetics preservative free or using natural products instead and their combination with the synthetics. Among the natural preservatives used, we can mention essential oils as a viable alternative. In this context, the present work aims to experimentally evaluate the antimicrobial potential of the Conobea scoparioides Cham. & Schltdl. essential oil, popularly known as pataqueira, the effect of its association with parabens and its effectiveness as a preservative in cosmetic bases. The essential oil composition was analyzed, indicating that it is composed mostly of terpenes, with thymol methyl ether (39.2%), thymol (33.8%) and -phelandrene (15.9%) as major compounds. The antimicrobial activity of essential oil and thymol was accessed through the minimum inhibitory concentration (MIC), whose results in µg/mL for essential oil and thymol were respectively: Staphylococcus aureus 650.70 and 284.90, Escherichia coli 721, 53 and 271.20, Pseudomonas aeruginosa 1748.00 and > 2,000, Burkholderia cepacia 833.03 and 1,077.70, Candida albicans 521.43 and 172.61 and Aspergillus brasiliensis 300 and 400. The synergistic effect of the association of essential oil with parabens was performed through a centroid simplex experimental design for a mixture of methylparaben, propylparaben and essential oil against the same microorganisms used in the antimicrobial activity evaluation The ideal concentrations obtained by statistical analysis for each component in µg/mL were: 1120 for methylparaben, 350 for propylparaben and 675 for essential oil. The effectiveness test of the preservative system in cosmetic formulation was carried out using the ideal concentrations plus two higher concentrations and one below the ideal. For all challenged microbial strains, the test result was a total reduction of the inoculated microbial load in the seven days of testing and no increase until the twenty-eighth day, which demonstrates the effectiveness of the association of essential oil with synthetic preservatives. C. scoparioides essential oil showed an important antimicrobial potential both alone and in association with parabens. These results demonstrated that it can be used to compose a preservative system for cosmetic formulations containing lower amounts of synthetics

Parabens disrupt non-canonical inflammasome activation.

Parabens are synthetic chemicals widely used as preservatives in cosmetics, pharmaceuticals, and foods. Although parabens, i.e., ethyl- and methyl-parabens, are considered relatively safe, study of possible health hazards has been undertaken due to the frequent exposure to parabens and their accumulation in the body. In this study, we elucidated the effect of parabens on inflammasome induction of inflammatory responses in innate immunity, such as interleukin (IL)-1ß maturation and gasdermin D (GSDMD)-mediating pyroptosis. Parabens attenuated the inflammatory responses to intracellular lipopolysaccharide (LPS) triggering of non-canonical (NC) inflammasome activation, but did not alter canonical inflammasome (i.e., NLRP3, NLRC4 and AIM2) responses. The NC inflammasome is assembled by the interaction of murine caspase (Casp)-11 (Casp4/5 in human) with cytosolic LPS, inducing endotoxin sepsis. Parabens selectively inhibited NC inflammasome activation in both human and murine macrophages and diminished the peritoneal IL-1ß production in LPS-injected mice. Parabens blocked the cleavage of GSDMD, Casp1, and Casp4, but did not change the expression of Casp11 or the activity of Casp1. Taken together, the results indicate that parabens could disrupt Gram-negative pathogen infection through the inhibition of NC inflammasome activation.

Promising anticancer activity of polysaccharides and other macromolecules derived from oyster mushroom (Pleurotus sp.): An updated review.

Cancer dominates among many causes of mortality worldwide. Traditional chemotherapeutic agents are powerful anti-cancer agents employed for treatment of this deadly disease. However, they are always associated with toxic side effects and immunosuppression making person more vulnerable to tumor relapse and fatalities. A promising alternative could be identification, isolation and transfer of naturally occurring bioactive macromolecules to the tumorigenic population. Oyster mushroom, a major source of nutraceuticals, belonging to class basidiomycetes of kingdom Mycota is known to have immense therapeutic properties. It is a reservoir of macromolecules like ß-glucan, α-glucan, resveratrol, concanavalin A, cibacron blue affinity protein, p-hydroxybenzoic acid, ergosterol, linoleic acid etc. that are responsible for mediating anti-tumor, immunomodulatory, antioxidant, and anti-diabetic roles. Various studies have shown that extracts derived from oyster mushroom is rich in polysaccharides like ß-glucan and other macro molecules which have an anti-proliferative effect against cancer cell lines, without harming the normal cells. This review presents a brief highlight of the work covering the overall significance of oyster mushroom in different types of cancer treatment. It also explores the immunomodulatory effects of polysaccharides, proteoglycans and polypeptides derived from oyster mushroom that boosts the immune system to overcome the limitation of traditional cancer therapies.

Mycobacterial para-Hydroxybenzoic Acid-Derivatives (pHBADs) and Related Structures Induce Macrophage Innate Memory.

Macrophages are key immune cells for combatting Mycobacterium tuberculosis. However, M. tuberculosis possesses means to evade macrophage bactericidal responses by, for instance, secretion of the immunomodulatory para-hydroxybenzoic acid derivatives (pHBADs). While these molecules have been implicated in inhibiting macrophage responses in an acute context, little is known about their ability to reprogram macrophages via induction of long-term innate memory. Since innate memory has been highlighted as a promising strategy to augment bactericidal immune responses against M. tuberculosis, investigating corresponding immune evasion mechanisms is highly relevant. Our results reveal for the first time that pHBAD I and related molecules (unmethylated pHBAD I and the hexose l-rhamnose) reduce macrophage bactericidal mechanisms in both the short- and the long-term. Moreover, we demonstrate how methyl-p-anisate hinders bactericidal responses soon after exposure yet results in enhanced pro-inflammatory responses in the long-term. This work highlights new roles for these compounds in M. tuberculosis pathogenesis.

Effects of Butylparaben Supplementation on In Vitro Development of Mouse Preantral Follicle.

We investigated whether butylparaben supplementation to the culture media negatively effects on in vitro development of mouse preantral follicle. The preantral follicles were isolated from the ovaries of 7-8-week-old mice and cultured in growth medium for 10 days and then in maturation medium for 2 days. During in vitro culture, butylparaben (0, 0.01, 0.1, 1.0, or 10 µM) was supplemented to the culture media. In the final spent media, the levels of 17ß-estradiol and anti-Müllerian hormone (AMH) were measured via enzyme-linked immunosorbent assay. In the final luteinized follicular cells, the mRNA levels of steroidogenic acute regulatory protein (StAR), superoxide dismutase 1 (Sod1), caspase 3 (Casp3), and extracellular signal-regulated kinase 1 (Erk1) were quantified via real-time reverse transcription-polymerase chain reaction. The metaphase II oocyte acquisition (per total oocyte) tended to decrease in the four butylparaben-supplemented groups, but not significant (26.8%, 23.2%, 21.4%, 15.1%, and 16.8%, respectively). The level of 17ß-estradiol and AMH tended to decrease in all butylparaben-supplemented groups, but statistically not significant. The expression level of StAR and Erk1 mRNA was significantly higher in all four butylparaben-supplemented groups, and a dose-dependent increment tendency was observed. Our findings suggest that butylparaben supplementation has largely no impact on in vitro development of mouse preantral follicle as well as 17ß-estradiol and AMH production. However, StAR, Sod1, Casp3, and Erk1 genes could be overexpressed in a certain concentration of butylparaben.

Transcriptome dynamics of alternative splicing events revealed early phase of apoptosis induced by methylparaben in H1299 human lung carcinoma cells.

Methylparaben is most frequently used as an antimicrobial preservative in pharmaceuticals and foods. Methylparaben has been subjected to toxicological studies owing to the increasing concern regarding its possible impact on the environment and human health. However, the cytotoxicity and underlying mechanisms of methylparaben exposure in human lung cells have not been explored. Here, we investigated the effect of methylparaben on cell cycle, apoptotic pathways, and changes in the transcriptome profiles in human lung cells. Our results demonstrate that treatment with methylparaben causes inhibition of cell growth. In addition, methylparaben induced S- and G2/M-phase arrest as a result of enhanced apoptosis. Transcriptome analysis using RNA-seq revealed that mRNA expression of ER stress- and protein misfolding-related gene sets was upregulated in methylparaben-treated group. RNA splicing- and maturation-related gene sets were significantly down-regulated by methylparaben treatment. Interestingly, RNA-seq analysis at the transcript level revealed that alternative splicing events, especially retained intron, were markedly changed by a low dose of methylparaben treatment. Altogether, these data show that methylparaben induces an early phase of apoptosis through cell cycle arrest and downregulation of mRNA maturation.

Enhancement of paraben-fungicidal activity by sulforaphane, a cruciferous vegetable-derived isothiocyanate, via membrane structural damage in Saccharomyces cerevisiae.

Parabens have been widely used as antimicrobial preservatives in cosmetics, pharmaceuticals, foods and beverages. Commonly, methyl-, ethyl-, propyl- and butylparaben are used independently or in combination to maintain the quality of industrial products, and they are considered to have low toxicity. However, recent evidence has suggested that parabens are toxic in mammalian cells, and parabens have been associated with allergic-contact dermatitis, breast cancer and changes in testosterone levels. Sulforaphane, a cruciferous vegetable-derived isothiocyanate, was effective in decreasing the growth inhibitory concentrations of ethyl-, propyl-, butyl- and methylparaben in the yeast Saccharomyces cerevisiae. The sulforaphane-enhanced fungicidal effects of methylparaben were deemed to be caused by drastic cell membrane damage and the leakage of internal substances, such as nucleotides, from S. cerevisiae cells. Moreover sulforaphane markedly decreased the minimum concentration of methyl- and ethylparaben required to inhibit the growth of various microbes, such as the pathogenic yeast that causes severe mycosis, Candida albicans; the filamentous fungi Aspergillus niger; and the Gram-negative bacterium Escherichia coli. Enhanced antimicrobial activity from the beneficial components of edible plants may increase paraben efficacy at low concentrations and minimize preservative-induced side effects in consumers. SIGNIFICANCE AND IMPACT OF THE STUDY: Sulforaphane, a natural and beneficial cruciferous vegetable-derived isothiocyanate, increased the ability of parabens to disrupt fungal cell membranes. Paraben-containing products have been reported to cause allergic contact dermatitis and drug hypersensitivity; therefore, methods to preserve organic products that may reduce the concentrations of parabens are both timely and necessary. In this study, we found that the combined antimicrobial effects of sulforaphane and parabens had the potential to reduce the paraben concentration needed to preserve organic products, thereby indicating that paraben toxicity may be reduced without affecting its activity as a preservative.

4-Hydroxybenzoic acid (4-HBA) enhances the sensitivity of human breast cancer cells to adriamycin as a specific HDAC6 inhibitor by promoting HIPK2/p53 pathway.

Breast cancer is reported a very complex disease along with heterogeneous morphological characteristics and unrelated clinical behavior, and is a leading cancer among female. Nevertheless, chemo-resistance is frequently observed. Adriamycin (ADM) is a always employed drug to treat clinical breast cancer. However, strong resistance to ADM limited its clinical efficacy. Deregulation of HDAC6 activity is linked to various diseases including cancer resulting in accumulating interest for developing HDAC6 inhibitors. In the present study, for the first time, we found that 4-Hydroxybenzoic acid (4-HBA), as histone deacetylase 6 (HDAC6) inhibitor, could successfully reverse ADM resistance in human breast cancer cells. 4-HBA significantly promoted the anticancer effect of ADM on apoptosis induction, as evidenced by the increased expressions of Caspase-3 and PARP cleavage, which was associated with the promotion p53 and homeodomain interacting protein kinase-2 (HIPK2) expressions in ADM-resistant breast cancer cells. Furthermore, the suppressive effect of ADM on drug-resistant breast cancer cells was accelerated by 4-HBA through increasing the number of cells distributed in G2/M phase of cell cycle arrest. Inhibiting HIPK2/p53 pathway could abolish 4-HBA/ADM co-treatment-induced apoptosis and G2/M cell cycle arrest. Importantly, HDAC6 expressions were also significantly down-regulated in ADM-resistance breast cancer cells co-treated with ADM and 4-HBA. Additionally, 4-HBA clearly potentiated the anticancer role of ADM in the MCF-7 breast cancer animal model with low toxicity. Therefore, 4-HBA could be applied as an effective HDAC6 inhibitor to reverse human breast cancer resistance. Herein, the 4-HBA and ADM combination might represent as a useful therapeutic strategy to prevent human breast cancer.

Condensed Tannins from Longan Bark as Inhibitor of Tyrosinase: Structure, Activity, and Mechanism.

In this study, the content, structure, antityrosinase activity, and mechanism of longan bark condensed tannins were evaluated. The findings obtained from mass spectrometry demonstrated that longan bark condensed tannins were mixtures of procyanidins, propelargonidins, prodelphinidins, and their acyl derivatives (galloyl and p-hydroxybenzoate). The enzyme analysis indicated that these mixtures were efficient, reversible, and mixed (competitive is dominant) inhibitor of tyrosinase. What's more, the mixtures showed good inhibitions on proliferation, intracellular enzyme activity and melanogenesis of mouse melanoma cells (B16). From molecular docking, the results showed the interactions between inhibitors and tyrosinase were driven by hydrogen bond, electrostatic, and hydrophobic interactions. In addition, high levels of total phenolic and extractable condensed tannins suggested that longan bark might be a good source of tyrosinase inhibitor. This study would offer theoretical basis for the development of longan bark condensed tannins as novel food preservatives and medicines of skin diseases.

Pseudoalteromonas haloplanktis TAC125 produces 4-hydroxybenzoic acid that induces pyroptosis in human A459 lung adenocarcinoma cells.

In order to exploit the rich reservoir of marine cold-adapted bacteria as a source of bioactive metabolites, ethyl acetate crude extracts of thirteen polar marine bacteria were tested for their antiproliferative activity on A549 lung epithelial cancer cells. The crude extract from Pseudoalteromonas haloplanktis TAC125 was the most active in inhibiting cell proliferation. Extensive bioassay-guided purification and mass spectrometric characterization allowed the identification of 4-hydroxybenzoic acid (4-HBA) as the molecule responsible for this bioactivity. We further demonstrate that 4-HBA inhibits A549 cancer cell proliferation with an IC50 value ≤ 1 µg ml-1, and that the effect is specific, since the other two HBA isomers (i.e. 2-HBA and 3-HBA) were unable to inhibit cell proliferation. The effect of 4-HBA is also selective since treatment of normal lung epithelial cells (WI-38) with 4-HBA did not affect cell viability. Finally, we show that 4-HBA is able to activate, at the gene and protein levels, a specific cell death signaling pathway named pyroptosis. Accordingly, the treatment of A549 cells with 4-HBA induces the transcription of (amongst others) caspase-1, IL1ß, and IL18 encoding genes. Studies needed for the elucidation of mode of action of 4-HBA will be instrumental in depicting novel details of pyroptosis.

A mixture of five endocrine-disrupting chemicals modulates concentrations of bisphenol A and estradiol in mice.

Most people in developed countries are exposed to multiple endocrine-disrupting synthetic chemicals. We previously showed that a single dose of triclosan, tetrabromobisphenol A (TBBPA), butyl paraben, propyl paraben, or di(2-ethylhexyl) phthalate elevated concentrations of bisphenol A (BPA) in mice. Here we investigated whether concurrent exposure to lower doses of these five chemicals could modulate concentrations of bisphenol A (BPA) or the natural estrogen, 17ß-estradiol (E2). CF1 mice were injected subcutaneously with 0.1 or 0.5 mg of one chemical, or a 0.5 mg mixture containing 0.1 mg of each of all five chemicals, then given dietary 50 µg kg-114C-BPA. The mixture elevated 14C-BPA concentrations in the lungs, muscle, uterus, ovaries, kidney, and blood serum of female mice. When administered alone, triclosan and TBBPA elevated 14C-BPA concentrations in the uterus, ovaries, and blood serum. In another experiment, CF1 mice were injected subcutaneously with the 0.5 mg mixture containing 0.1 mg of all five chemicals, then E2 was measured in urine 2-12 h later. The mixture elevated E2 at 8 h after injection in female mice. No treatments significantly altered concentrations of 14C-BPA or E2 in male mice. These data show that these endocrine-disrupting chemicals interact in vivo, magnifying one another's effects, consistent with inhibition of enzymes that are critical for estrogen metabolism. These findings highlight the importance of considering exposure to multiple chemicals when assessing health outcomes and determining regulatory exposure limits.

Interference of Paraben Compounds with Estrogen Metabolism by Inhibition of 17ß-Hydroxysteroid Dehydrogenases.

Parabens are effective preservatives widely used in cosmetic products and processed food, with high human exposure. Recent evidence suggests that parabens exert estrogenic effects. This work investigated the potential interference of parabens with the estrogen-activating enzyme 17ß-hydroxysteroid dehydrogenase (17ß-HSD) 1 and the estrogen-inactivating 17ß-HSD2. A ligand-based 17ß-HSD2 pharmacophore model was applied to screen a cosmetic chemicals database, followed by in vitro testing of selected paraben compounds for inhibition of 17ß-HSD1 and 17ß-HSD2 activities. All tested parabens and paraben-like compounds, except their common metabolite p-hydroxybenzoic acid, inhibited 17ß-HSD2. Ethylparaben and ethyl vanillate inhibited 17ß-HSD2 with IC50 values of 4.6 ± 0.8 and 1.3 ± 0.3 µM, respectively. Additionally, parabens size-dependently inhibited 17ß-HSD1, whereby hexyl- and heptylparaben were most active with IC50 values of 2.6 ± 0.6 and 1.8 ± 0.3 µM. Low micromolar concentrations of hexyl- and heptylparaben decreased 17ß-HSD1 activity, and ethylparaben and ethyl vanillate decreased 17ß-HSD2 activity. However, regarding the very rapid metabolism of these compounds to the inactive p-hydroxybenzoic acid by esterases, it needs to be determined under which conditions low micromolar concentrations of these parabens or their mixtures can occur in target cells to effectively disturb estrogen effects in vivo.

Combination of essential oil compounds and phenolic acids against Escherichia coli O157:H7 in vitro and in dry-fermented sausage production.

Escherichia coli O157:H7 is a foodborne pathogen that causes hemorrhagic colitis and hemolytic uremic syndrome. The low dose of infection and severity of the disease represent a concern to public health. Natural compounds have been widely applied as food additives to replace synthetic preservatives. The aim of this study was to determine the efficiency of essential oil compounds (EOCs) in combination with phenolic acids (PA) in vitro and in dry-fermented sausage production. Minimum Inhibitory Concentration (MIC) and Fractional Inhibitory Concentration index (FICindex) were determined for a 5-strain mixture of E. coli O157:H7. Batches of sausage tainted with E. coli O157:H7 were produced using Pediococcus pentosaceus UM 116P and Staphylococcus carnosus UM 123M as starter cultures. The best combination of EOCs and PAs found in vitro was used as an additive. Chemical-physical and microbiological analyses were evaluated weekly from day 0 to 35 after production. Sensory evaluation (texture, odor, flavor, appearance and overall evaluation) of E. coli-free sausages was conducted using a 9-point hedonic scale with 56 untrained volunteers. The MIC values of allyl isothiocyanate (AITC), carvacrol (CAR), ferulic acid (FA), o-coumaric acid (CA) and p-hydroxybenzoic acid (AHB) were, respectively, 0.25; 1.3; 5.12; 18.27; and 37mM. AITC combined with CA had a synergistic effect (FICindex=0.25) and together they were applied in the production of dry fermented sausage at concentrations of 10× FIC and 20× FIC. Aw had no significant difference among treatments, whereas the pH of 10× FIC and 20× FIC were higher than the control. E. coli O157:H7 was reduced by >5logCFU/g with 20× FIC after 21d, and by 2.8logCFU/g with 10× FIC after 35d. Sensory analysis showed that the combination of AITC and ο-coumaric acid in both treatments presented lower scores in the 5 categories when compared to the control, but none of the parameters received a negative score. This study demonstrated that the combination of AITC and ο-coumaric acid at 20× FIC reduced E. coli O157:H7 in compliance with the North American legislation, but adjustments in the dose are necessary to improve the sensory characteristics of the final product.

A study on the in vitro percutaneous absorption of silver nanoparticles in combination with aluminum chloride, methyl paraben or di-n-butyl phthalate.

Some reports indicate that the silver released from dermally applied products containing silver nanoparticles (AgNP) (e.g. wound dressings or cosmetics) can penetrate the skin, particularly if damaged. AgNP were also shown to have cytotoxic and genotoxic activity. In the present study percutaneous absorption of AgNP of two different nominal sizes (Ag15nm or Ag45nm by STEM) and surface modification, i.e. citrate or PEG stabilized nanoparticles, in combination with cosmetic ingredients, i.e. aluminum chloride (AlCl3), methyl paraben (MPB), or di-n-butyl phthalate (DBPH) was assessed using in vitro model based on dermatomed pig skin. The inductively coupled plasma mass spectrometry (ICP-MS) measurements after 24h in receptor fluid indicated low, but detectable silver absorption and no statistically significant differences in the penetration between the 4 types of AgNP studied at 47, 470 or 750µg/ml. Similarly, no significant differences were observed for silver penetration when the AgNP were used in combinations with AlCl3 (500µM), MPB (1250µM) or DBPH (35µM). The measured highest amount of Ag that penetrated was 0.45ng/cm2 (0.365-0.974ng/cm2) for PEG stabilized Ag15nm+MPB.

A Novel Class of Natural FXR Modulators with a Unique Mode of Selective Co-regulator Assembly.

The farnesoid X receptor (FXR) is an important target for drug discovery. Small molecules induce a conformational change in FXR that modulates its binding to co-regulators, thus resulting in distinct FXR functional profiles. However, the mechanisms for selectively recruiting co-regulators by FXR remain elusive, partly because of the lack of FXR-selective modulators. We report the identification of two natural terpenoids, tschimgine and feroline, as novel FXR modulators. Remarkably, their crystal structures uncovered a secondary binding pocket important for ligand binding. Further, tschimgine or feroline induced dynamic conformational changes in the activation function 2 (AF-2) surface, thus leading to differential co-regulator recruiting profiles, modulated by both hydrophobic and selective hydrogen-bond interactions unique to specific co-regulators. Our findings thus provide a novel structure template for optimization for FXR-selective modulators of clinical value.

Transformed Root Extract of Leonurus sibiricus Induces Apoptosis through Intrinsic and Extrinsic Pathways in Various Grades of Human Glioma Cells.

This study determines the influence of transformed root (TR) extract of Leonurus sibiricus L. on various grades (I-III) of human glioma cells derived from patients. This plant occurs in southern Asia and Siberia and is widely used as a medicinal plant with various biological activities. Chromatographic profile of TR extract have revealed the presence of various polyphenolic compounds (4-hydroxybenzoic acid, gentisic acid, vanilic acid, 1,3-dicaffeoylquinic acid, α-resorcylic acid). We found TR root extract to have antiproliferative activity on glioma cells after 24 h of treatment. TR root extract induces apoptosis on various grades (I-III) of human glioma cells by the generation of reactive oxygen species (ROS) along with concurrent loss of mitochondrial membrane potential, enhanced S and G2/M phases of the cell cycle, and altered mRNA levels of Bax, Bcl-2, p53, Cas-3, Cas-8 and Cas-9 factors involved in apoptosis. This work for the first time demonstrate that TR extract from L. sibiricus root has the potential to activate apoptosis in grade I-III human glioma cells through the intrinsic and extrinsic pathways.

Essential Oil from Sweet Potato Vines, a Potential New Natural Preservative, and an Antioxidant on Sweet Potato Tubers: Assessment of the Activity and the Constitution.

Pathogenic fungi and oxidation are the major factors that cause the deterioration of sweet potatoes and also cause the loss of quality that makes consumption unsafe. In the present study, the in vitro results demonstrate that the essential oil from sweet potato vines exhibits significantly enhanced activity compared to that of the control. Furthermore, the essential oil can actively inhibit the growth of some common microorganisms inducing pathogenic bacteria and fungi (inhibition rates above 50% at low concentrations). A total of 31 constituents were identified using GC-MS and confirmed that linalool and p-hydroxybenzoic acid are the major active ingredients. The experiment involving actual tubers showed that the essential oil could retains its quality and effectiveness again the fungus disease. This suggests that it could be used in the food industry to increase the shelf life of stored produce (tubers) to ensure food safety without the use of additives or preservatives.

Phenolic constituents from Wissadula periplocifolia (L.) C. Presl. and anti-inflammatory activity of 7,4'-di-O-methylisoscutellarein.

This study reports the first phenolics from Wissadula genus (Malvaceae) and the anti-inflammatory activity of 7,4'-di-O-methylisoscutellarein. Using chromatographic methods, five phenolic compounds were isolated from aerial parts of Wissadula periplocifolia (L.) C. Presl. The compounds were identified as 4-hydroxybenzoic acid, 3-hydroxybenzoic acid, trans-cinnamic acid, tamgermanetin and 7,4'-di-O-methylisoscutellarein using spectroscopic methods. The flavone 7,4'-di-O-methylisoscutellarein showed anti-inflammatory activity by inhibiting neutrophils recruitment in a mice model of pleurisy and by decreasing significantly the production of cytokines IL-1ß and TNF-α.

First paraben substituted cyclotetraphosphazene compounds and DNA interaction analysis with a new automated biosensor.

Cancer, as one of the leading causes of death in the world, is caused by malignant cell division and growth that depends on rapid DNA replication. To develop anti-cancer drugs this feature of cancer could be exploited by utilizing DNA-damaging molecules. To achieve this, the paraben substituted cyclotetraphosphazene compounds have been synthesized for the first time and their effect on DNA (genotoxicity) has been investigated. The conventional genotoxicity testing methods are laborious, take time and are expensive. Biosensor based assays provide an alternative to investigate this drug/compound DNA interactions. Here for the first time, a new, easy and rapid screening method has been used to investigate the DNA damage, which is based on an automated biosensor device that relies on the real-time electrochemical profiling (REP™) technology. Using both the biosensor based screening method and the in vitro biological assay, the compounds 9 and 11 (propyl and benzyl substituted cyclotetraphosphazene compounds, respectively), have resulted in higher DNA damage than the others with 65% and 80% activity reduction, respectively.