Myra Villareal

Many ovarian cancers initially respond well to chemotherapy, but often become drug-resistant after several years. Therefore, analysis of drug resistance mechanisms and overcoming resistance are urgently needed. Paclitaxel is one of the first-choice and widely-used drugs for ovarian cancer, but like most drugs, drug resistance is observed in subsequent use. RSK4 is known as a tumor-suppressor, however, it has increasingly been reported to lead to drug resistance. Here, we found that RSK4 expression was elevated in paclitaxel-resistant ovarian cancer cells using DNA microarray, quantitative real-time PCR, and western blotting analysis. We examined the contribution of RSK4 to paclitaxel resistance and found that paclitaxel sensitivity was restored by RSK inhibitor co-treatment. We analyzed the mechanism by which resistance is developed when RSK4 level is elevated, and accelerated phosphorylation of the downstream translation factor eIF4B was discovered. In the Kaplan-Meier plot, the overall survival time was longer with RSK4 high, supporting its role as a tumor suppressor, as in previous findings, but the tendency was reversed when focusing on paclitaxel treatment. In addition, RSK4 levels were higher in non-responders than in responders in the ROC plotter. Finally, external expression of RSK4 in ovarian cancer cells increased the cell viability under paclitaxel treatment. These findings suggest that RSK4 may contribute to paclitaxel resistance, and that co-treatment with RSK4 inhibitors is effective treatment of paclitaxel-resistant ovarian cancer in which RSK4 is elevated.

Plant saponins are abundant and diverse natural products with a great potential for use in drug-discovery research. Here, we evaluated extracts of saponins-rich fractions of argan leaves and argan oil extraction byproducts (shell, pulp, press cake) for their effect on melanogenesis. Results show that from among the samples tested, only the saponins-rich fraction from leaves (ALS) inhibited melanin production in B16 murine melanoma (B16) cells. The mechanism of the melanogenesis inhibition was elucidated by determining the protein and mRNA expression of melanogenesis-associated enzymes tyrosinase (TYR), tyrosinase-related protein 1 (TRP1), and dopachrome tautomerase (DCT), and microphthalmia-associated transcription factor (MITF), and performing DNA microarray analysis. Results showed that 10 µg/mL ALS significantly inhibited melanogenesis in B16 cells and human epidermal melanocytes by 59% and 48%, respectively, without cytotoxicity. The effect of ALS on melanogenesis can be attributed to the decrease in TYR, TRP1, and MITF expression at the protein and mRNA levels. MITF inhibition naturally led to the downregulation of the expression of Tyr and Trp1 genes. Results of the DNA microarray analysis revealed the effect on melanogenesis-associated cAMP and Wnt signaling pathways’ genes. The results of this study suggest that ALS may be used in cosmeceuticals preparations for hyperpigmentation treatment.

The beneficial effect on health of argan oil is recognized worldwide. We have previously reported that the cake that remains after argan oil extraction (argan press-cake or APC) inhibits melanogenesis in B16 melanoma cells in a time-dependent manner without cytotoxicity. In this study, the global gene expression profile of B16 melanoma cells treated with APC extract was determined in order to gain an understanding of the possible mechanisms of action of APC. The results suggest that APC extract inhibits melanin biosynthesis by down-regulating microphthalmia-associated transcription factor (Mitf) and its downstream signaling pathway through JNK signaling activation, and the inhibition of Wnt/beta-catenin and cAMP/PKA signaling pathways. APC extract also prevented the transport of melanosomes by down-regulating Rab27a expression. These results suggest that APC may be an important natural skin whitening product and pharmacological agent used for clinical treatment of pigmentary disorders.

Hair loss is a distressing condition that may not be life-threatening but has an indisputable impact on psychological well-being of an individual. African plant resources have a great potential for use in hair growth promotion. Here, the effect of tara tannin from tara (Caesalpinia spinosa) pods on hair growth promotion was investigated in vitro using hair follicle dermal papilla cells (HFDPC). The noncytotoxic concentration of tara tannin was determined by subjecting HFDPCs to cytotoxicity assay. Then, ATP production was evaluated and quantitative polymerase chain reaction (qPCR) of hair growth promotion molecular markers was performed to determine the promotion effect on hair growth. Results showed that 5 mu M tara tannin stimulated HFPDC proliferation, accompanied by an increase in ATP production. Fluorescent staining revealed an increase in ss-catenin in tara-tannin-treated cells. QPCR results confirmed that 5 mu M tara tannin upregulated the expression of ss-catenin (CTNBB), alkaline phosphatase (tissue-nonspecific isozyme) (ALPL), neural cell adhesion molecule 1 (NCAM1), and fibroblast growth factor 1 (FGF1) in HFDPCs. These genes' expression was upregulated in dermal papilla

This study identified anserine and anserine/carnosine in chicken breast of Thai native chicken (TNC; 100% Thai native), Thai synthetic chicken (TSC; 50% Thai native), and Thai native crossbred chicken (TNC crossbred; 25% Thai native) compared with commercial broiler chicken (BR; 0% Thai native) using nuclear magnetic resonance (NMR) spectroscopy and the effect on antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl assay (DPPH). We conducted experiments with a completely randomized design and explored principal components analysis (PCA) and orthogonal projection to latent structure-discriminant analysis (OPLS-DA) to identify the distinguishing metabolites and relative concentrations from 1H NMR spectra among the groups. The relative concentrations and antioxidant properties among the groups were analyzed by analysis of variance (ANOVA) using the general linear model (GLM). This study revealed seven metabolites alanine, inositol monophosphate (IMP), inosine, and anserine/carnosine, lactate, anserine, and creatine. Lactate, anserine, and creatine were major components. In terms of PCA, the plots can distinguish BR from other groups. OPLS-DA revealed that anserine and anserine/car

The use of natural products for the regulation of skin pigmentation is gaining popularity. In the present study, we evaluated the effect of argan leaves extract (ALE) on melanogenesis in B16 melanoma cells, determined its antioxidant activity, then quantified and identified its phenolic components. B16 cells were treated with various concentrations of ALE, then the cell viability and proliferation were assessed using MTT assay while the melanin content was determined using spectrophotometric methods. The expression level of tyrosinase (TYR), tyrosinase related protein-1 (TRP-1) and dopachrome tautomerase (DCT) was evaluated by Western blotting. The antioxidant activity of ALE was investigated using four different assays while UPLC-ESI-HRMS analysis was used to characterize the ALE phenolic profile. Fourteen phenolic compounds were identified, of which six are reported for the first time to be present in ALE. ALE treatment increases the melanin content of B16 cells in a dose-dependent manner without cytotoxicity. This was revealed by the observed ALE-increased expression level of TYR, DCT, and TRP-1. These bioactivities may be mainly attributed to its high flavonoids content. Argan

Numerous studies have been carried out on the bioactive properties of hydroxytyrosol (HT) in olive oils (OOs), although there are few reports comparing anti-inflammatory activity among different olive varieties or regions of production. The purpose of this study was to investigate the in vitro inflammatory action of HT in extracts of four OO varieties in the Languedoc region of the French Mediterranean. Factors other than cultivar were eliminated, which enabled unambiguous varietal differences to be identified. Purified extracts of OO were obtained using an optimized solid-phase extraction procedure by which only polar compounds were recovered. High performance liquid chromatography-photodiode array detection-tandem mass spectrometry was used to identify and quantify HT and oleacein in the extracts. The total polyphenol concentration ranged from 93.00mg gallic acid equivalent/kg OO for Picholine to 27mg gallic acid equivalent for Verdale OOs. The concentrations of HT in Picholine, Olivere, and Lucques varieties were 25.3, 18.8 and 12.1mg/kg, respectively, whereas the concentration of HT in Verdale OOs was less, 1mg/kg. The in vitro anti-inflammatory response of purified OO extracts

Melanoma is the most dangerous form of skin cancer with a very poor prognosis. Melanoma develops when unrepaired DNA damage causes to skin cells to multiply and form malignant tumors. The current therapy is limited by the highly ability of this disease to metastasize rapidly. Plumbagin is a naphthoquinone (5-hydroxy-2-methyl-1, 4-naphthoquinone), isolated from the roots of medicinal plant Plumbago zeylanica, and it is widely present in Lawsonia inermis L. It has been shown that plumbagin has an anti-proliferative and anti-invasive activities in various cancer cell lines; however, the anti-cancer and anti-metastatic effects of plumbagin are largely unknown against melanoma cells. In this study, we evaluated the effect of plumbagin on B16F10 murine melanoma cells . Plumbagin decreased B16F10 cell viability as well as the cell migration, adhesion, and invasion. The molecular mechanism was studied, and plumbagin downregulated genes relevant in MAPK pathway, matrix metalloproteinases (MMP's), and cell adhesion. Furthermore, plumbagin elevated the expression of apoptosis and tumors suppressor genes, and genes significant in reactive oxygen species (ROS) response. Taken together, our findings suggest that plumbagin has an anti-invasion and anti-metastasis effect on melanoma cancer cells by acting on MAPK pathway and its related genes.

The hair follicle undergoes a regular cycle composed of three phases: anagen, catagen, and telogen. The life of follicular melanocytes is totally linked to the hair cycle; and during anagen or the growth phase, the melanocytes are active and produce the melanin responsible of hair shaft pigmentation. Various signaling pathways regulate the hair growth cycle and, therefore, the pigmentation; we distinguish the Wnt/β-catenin signaling pathway as it plays a major role in the development, growth, and proliferation of the melanocytes and the activation of melanogenesis enzymes and the related transcription factor. In this study, 3,4,5-tri-O-caffeoylquinic acid (TCQA), a caffeoylquinic acid derivative, stimulated the pigmentation in C3H mouse hair follicle, in human melanocytes, and B16F10 melanoma cells. An enhancement in pigmentation associated genes was observed upon TCQA treatment in vivo and in vitro. Interestingly, the expression of β-catenin was remarkably upregulated in mouse treated skin and in pigment cell lines. Moreover, TCQA upregulated CTNNB1 expression after inhibition in human melanocytes. Taken together, this study suggests that TCQA triggered β-catenin activation to enhance the pigmentation during the anagen phase of the hair cycle.

We have previously reported that argan oil and argan press-cake from the kernels of Argania spinosa have an anti-melanogenesis effect. Here, the effect of argan fruit shell ethanol extract (AFSEE) on melanogenesis in B16F10 cells was determined, and the mechanism underlying its effect was elucidated. The proliferation of AFSEE-treated B16F10 cells was evaluated using the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay, while the melanin content was quantified using a spectrophotometric method. The expression of melanogenesis-related proteins was determined by Western blot and real-time PCR, while global gene expression was determined using a DNA microarray. In vitro analysis results showed that the melanin content of B16F10 cells was significantly increased by AFSEE, without cytotoxicity, by increasing the melanogenic enzyme tyrosinase (TRY), tyrosinase related-protein 1 (TRP1), and dopachrome tautomerase (DCT) protein and mRNA expression, as well as upregulating microphthalmia-associated transcription factor (MITF) expression through mitogen-activated protein kinases (MAPKs) extracellular signal-regulated kinase (ERK) and p38, and the cyclic adenosine monophosphate (cAMP) signaling pathway, as indicated by the microarray analysis results. AFSEE's melanogenesis promotion effect is primarily attributed to its polyphenolic components. In conclusion, AFSEE promotes melanogenesis in B16F10 cells by upregulating the expression of the melanogenic enzymes through the cAMP-MITF signaling pathway.AFSEE may be used as a cosmetics product component to promote melanogenesis, or as a therapeutic against hypopigmentation disorders.

The hair follicle is a complex structure that goes through a cyclic period of growth (anagen), regression (catagen), and rest (telogen) under the regulation of several signaling pathways, including Wnt/ β-catenin, FGF, Shh, and Notch. The Wnt/β-catenin signaling is specifically involved in hair follicle morphogenesis, regeneration, and growth. β-catenin is expressed in the dermal papilla and promotes anagen induction and duration, as well as keratinocyte regulation and differentiation. In this study, we demonstrated the activation of β-catenin by a polyphenolic compound 3,4,5-tri--caffeoylquinic acid (TCQA) in mice model and in human dermal papilla cells to promote hair growth cycle. A complete regrowth of the shaved area of C3H mice was observed upon treatment with TCQA. Global gene expression analysis using microarray showed an upregulation in hair growth-associated genes. Moreover, the expression of β-catenin was remarkably upregulated and . These findings suggest that β-catenin activation by TCQA promoted the initiation of the anagen phase of the hair cycle.

BACKGROUND: Melanoma is one of the most invasive and aggressive types of cancer with a very poor prognosis. Surgery remains the most efficient treatment prior melanoma invasion and metastasis formation. However, therapy becomes a challenge once the cancer cells colonized other tissues. At present, there are two main classes of therapies acting with a certain efficiency on metastatic melanoma: immune check point inhibitors (anti-PD1/PDL1) and targeted therapy such as Vemurafenib. Unfortunately, these therapies are not fully responsive, induce resistance and/or generate unwanted side effects. In this respect, it is important to continue to discover new cancer therapeutics. Here, we show that daphnane diterpenes type of compounds can prevent melanoma metastasis by inhibiting metastasis-associated matrix metalloproteinases expression without cytotoxicity. METHODS: Evaluation of the anti-metastasis effect of daphnane diterpenes-rich Thymelaea hirsuta extract (TH) and its bioactive component gnidilatidin was carried out in vitro using B16 murine melanoma cells and in vivo using male C57BL/6 J mice. Global gene expression in B16 cells was done using DNA microarray, validated using real-time PCR, to further understand the effect of daphnane diterpenes, specifically daphnane diterpenoid gnidilatidin. RESULTS: Oral administration of daphnane diterpenes-rich Thymelaea hirsuta extract (TH) suppressed MMP2 and MMP9 expression, decreasing lung tumor in mice injected with B16 murine melanoma cells. Validation of these observations in vitro showed reduced B16 cells migration, adhesion, and invasion. Results of microarray analysis of B16 cells treated with daphnane diterpenoid gnidilatidin from TH revealed an upregulation of tumor suppressor Egr1 while inhibiting metastasis-associated genes Id2 and Sytl2 expression. A downregulation of the melanoma oncogene microphthalmia-associated transcription factor (Mitf) was observed, and most likely caused by the inhibition of Id2, a gene that regulated HLH transcription factors such as MITF and also reported to promote tumor cell migration and invasion. CONCLUSIONS: Daphnane diterpenes have inhibitory effect on the metastatic potential of B16 melanoma cells, and the results of this study provided evidence for their potential for use in the prevention and inhibition of melanoma metastasis.

BACKGROUND: Melanoma is one of the most invasive and aggressive types of cancer with a very poor prognosis. Surgery remains the most efficient treatment prior melanoma invasion and metastasis formation. However, therapy becomes a challenge once the cancer cells colonized other tissues. At present, there are two main classes of therapies acting with a certain efficiency on metastatic melanoma: immune check point inhibitors (anti-PD1/PDL1) and targeted therapy such as Vemurafenib. Unfortunately, these therapies are not fully responsive, induce resistance and/or generate unwanted side effects. In this respect, it is important to continue to discover new cancer therapeutics. Here, we show that daphnane diterpenes type of compounds can prevent melanoma metastasis by inhibiting metastasis-associated matrix metalloproteinases expression without cytotoxicity. METHODS: Evaluation of the anti-metastasis effect of daphnane diterpenes-rich Thymelaea hirsuta extract (TH) and its bioactive component gnidilatidin was carried out in vitro using B16 murine melanoma cells and in vivo using male C57BL/6 J mice. Global gene expression in B16 cells was done using DNA microarray, validated using real-time PCR, to further understand the effect of daphnane diterpenes, specifically daphnane diterpenoid gnidilatidin. RESULTS: Oral administration of daphnane diterpenes-rich Thymelaea hirsuta extract (TH) suppressed MMP2 and MMP9 expression, decreasing lung tumor in mice injected with B16 murine melanoma cells. Validation of these observations in vitro showed reduced B16 cells migration, adhesion, and invasion. Results of microarray analysis of B16 cells treated with daphnane diterpenoid gnidilatidin from TH revealed an upregulation of tumor suppressor Egr1 while inhibiting metastasis-associated genes Id2 and Sytl2 expression. A downregulation of the melanoma oncogene microphthalmia-associated transcription factor (Mitf) was observed, and most likely caused by the inhibition of Id2, a gene that regulated HLH transcription factors such as MITF and also reported to promote tumor cell migration and invasion. CONCLUSIONS: Daphnane diterpenes have inhibitory effect on the metastatic potential of B16 melanoma cells, and the results of this study provided evidence for their potential for use in the prevention and inhibition of melanoma metastasis.

[SCOPE] L-citrulline has recently been reported as a more effective supplement for promoting intracellular NO production compared to L-arginine. Here, the effect of L-citrulline on skeletal muscle and its influence on exercise performance were investigated. The underlying mechanism of its effect, specifically on the expression of skeletal muscle peroxisome proliferator-activated receptor-gamma coactivator-1α (PGC-1α), was also elucidated. [METHODS AND RESULTS] Six-week-old ICR mice were orally supplemented with L-citrulline (250 mg kg ) daily, and their performance in weight-loaded swimming exercise every other day for 15 days, was evaluated. In addition, mice muscles were weighed and evaluated for the expression of PGC-1α and PGC-1α-regulated genes. Mice orally supplemented with L-citrulline had significantly higher gastrocnemius and biceps femoris muscle mass. Although not statistically significant, L-citrulline prolonged the swimming time to exhaustion. PGC-1α upregulation was associated with vascular endothelial growth factor α (VEGFα) and insulin-like growth factor 1 (IGF1) upregulation. VEGFα and IGF1 are important for angiogenesis and muscle growth, respectively, and

Chronic knee joint pain is common in the elderly and associated with poor quality of life. This study, an open-label clinical trial, aimed to examine how the intake on a daily basis of maslinic acid-containing product (30 mg maslinic acid) on 29 elderly residents (mean 70.7 ± 10.1 years) of Nakajima Island, Ehime, Japan. Study participants consumed 10 g jelly containing maslinic acid daily for 16 weeks and at 0 (baseline), 4, 8, 12 and 16 weeks, assessed for health-related quality of life (Short Form-8) and knee pain score (Japanese Knee Osteoarthritis Measure). After 16 weeks, the physical quality of life, more specifically, the level of Bodily Pain and Physical Component Summary, but not mental quality of life, was significantly improved by maslinic acid intake. Furthermore, maslinic acid intake significantly decreased the Japanese Knee Osteoarthritis Measure at week 8 and tended to decrease Visual Analogue Scale score at weeks 4 and 16. These results suggest that consumption of maslinic acid has a protective effect against chronic knee pain in elderly residents in a community where knee pain causes high quality of life burden.

Consumption of olives (Olea europaea L.) is associated with a low incidence of inflammation-related diseases. Olive fruit is rich in bioactive pentacyclic triterpenoids, mainly maslinic acid. This study, a randomized, double-blind, and placebo-controlled trial, examined the effects of an orally administered maslinic acid supplement, olive fruit extract, on 20 middle-aged and elderly volunteers with mild knee joint pain. Each subject (58 ± 7 years) received either olive fruit extract, containing 50 mg maslinic acid (n = 12), or placebo (n = 8) daily for 12 weeks and evaluated for pain and physical functions as primary outcome measures. Secondary outcome measures included body composition and inflammatory biomarkers in serum. Although both groups exhibited improved pain visual analogue scale score and quality of life after supplementation, symptoms were better in the maslinic acid group than in the placebo group. After 12 weeks, maslinic acid group exhibited significant decrease in body weight and body mass index suggesting that maslinic acid affected the weight of volunteers with mild knee joint pain. Therefore, olive products containing maslinic acid may be useful as a new preventive and therapeutic food ingredient for arthritic diseases. Since this clinical study is a preliminary study, it was not registered in a publicly accessible database.

The harsh environment of semi-arid and arid regions induces plants in these regions to produce numerous bioactive compounds with therapeutic or medicinal properties. An initial study of extracts from arid land plants Cymbopogon schoenanthus, Crithmum maritimum, Rhanterium suaveolens, and Artemisia herba-alba evaluated their effects on Type I allergic reactions and melanin biosynthesis using RBL-2H3 basophilic cells and B16 murine melanoma cells, respectively. Plant extracts were not cytotoxic at low concentrations. β-hexosaminidase release inhibition assay indicated that extracts significantly inhibited mast cell degranulation. Melanin assay results showed significant melanin biosynthesis regulatory effects in B16 cells. Further studies are being undertaken to understand the mechanism underlying the observed effects.

Hematopoietic stem cells (HSCs) transplantation and/or the infusion of hematopoietic cells are currently the best alternative for the treatment of a wide variety of hematological disorders. Novel modulators of HSCs fate continue therefore to be identified in an attempt to optimize the in vitro culture of these cells. In this study, main olive leaf components, oleuropein, apigenin 7-glucoside, and luteolin 7-glucoside, and their combination, were investigated for their effect on the viability, self-renewal and differentiation of CD34+ hematopoietic cells. High concentrations of up to 50 μM of these olive leaf phytochemicals, as well as their combination, did not decrease CD34+ cell viability suggesting that these compounds are not cytotoxic on HSCs. Flow cytometric analysis revealed a decrease in the expression of CD34 on the cell surface of HSCs after 9 days of treatment with these compounds indicating a decrease in the proportion of the stem population. Results of the colony-forming unit assay showed an increase in the number of different colonies following treatment with olive leaf phytochemicals. These findings suggest that olive leaf phytochemicals, used alone or in combinatio