New synthetic raloxifen-like di(hetero)arylamines induce apoptosis and inhibit the estrogen receptor in breast cancer cells

Breast cancer is the most common form of cancer in women in the western world, and in spite of some decline in death rates in recent years it is still the second most common cause of death from cancer, in women (1). For more than three decades, the estrogen receptor (ER) has been the most important biomarker of breast cancer, largely due to the substantial benefit that endocrine therapy provides in the treatment of ER positive tumors, in women of all ages (2). Endocrine therapies currently available include selective estrogen receptor modulators (SERMs), like tamoxifen and raloxifene (3). Raloxifene acts as an estrogen antagonist in the breast by competitive binding to the ER, inhibiting estrogen-induced breast tissue proliferation and preventing the growth of mammary tumors. In addition, it shows no increase in incidence of endometrial cancer, which is an advantage compared to tamoxifen (4). The successes of these endocrine therapies, however, are often limited. So, it is important to continue searching for new strategies and/or drugs that overcome resistance problems and that can be potent enough with fewer adverse effects. In this work, we studied the effect of two new synthetic di(hetero)arylamines, named MJQ2 and MJQ3 (which have in common with raloxifene a benzothiophene ring), in cell proliferation and apoptosis of two different cell lines from breast cancer: MCF-7 (ER positive) and MDA-MB-231 (ER negative). Our results showed that both diarylamines induce apoptosis without significant necrosis (evaluated by Hoechst-PI staining), at the IC50 concentration that inhibits cell proliferation (evaluated by the SRB assay). The results obtained with TMRM, a marker of mitochondrial membrane potential, suggest that mitochondrial disruption can be involved in this apoptotic process. These effects are more pronounced in the MCF-7 cell line (ER positive), suggesting that the presence of the ER might be important in the response to these compounds. Confirmation of their interaction with the ER was obtained in the E-Screen assay, where a clear antagonism of the proliferative effects of the hormone 17β-estradiol was observed with both compounds, at non-toxic concentrations. The overall results suggest that these new synthetic “raloxifene-like” drugs might have potential to be further developed as alternative hormonal or adjuvant therapies for breast cancer.Fundação para a Ciência e a Tecnologia (FCT) research project PTDC/QUI/68382/2006 (FCOMP-01-0124-FEDER 007441)

Angolan cymbopogon citratus used for therapeutic benefits: nutritional composition and influence of solvents in phytochemicals content and antioxidant activity of leaf extracts

Folk medicine is a relevant and effective part of indigenous healthcare systems which are, in practice, totally dependent on traditional healers. An outstanding coincidence between indigenous medicinal plant uses and scientifically proved pharmacological properties of several phytochemicals has been observed along the years. This work focused on the leaves of a medicinal plant traditionally used for therapeutic benefits (Angolan Cymbopogon citratus), in order to evaluate their nutritional value. The bioactive phytochemical composition and antioxidant activity of leaf extracts prepared with different solvents (water, methanol and ethanol) were also evaluated. The plant leaves contained ~60% of carbohydrates, protein (~20%), fat (~5%), ash (~4%) and moisture (~9%). The phytochemicals screening revealed the presence of tannins, flavonoids, and terpenoids in all extracts. Methanolic extracts also contained alkaloids and steroids. Several methods were used to evaluate total antioxidant capacity of the different extracts (DPPH; NO; and H2O2 scavenging assays, reducing power, and FRAP). Ethanolic extracts presented a significantly higher antioxidant activity (p < 0.05) except for FRAP, in which the best results were achieved by the aqueous extracts. Methanolic extracts showed the lowest radical scavenging activities for both DPPH; and NO; radicals

Evaluation of antioxidant and antimicrobial properties of the Angolan Cymbopogon Citratus essential oil with a view to its utilization as food biopreservative

It was studied the chemical composition, antioxidant, antibacterial and antifungal properties of the essential oil obtained from the Cymbopogon citratus of Angolan origin. Its major constituents analyzed by GC-MS were α-citral (40.55%), β-citral (28.26%), myrcene (10.50%) and geraniol (3.37%). The essential oil antioxidant capacity was statistically identical to that of synthetic antioxidants (DPPH IC50 of 41.7 μg/ml) and superior to that of extracts obtained from fresh leaves of the plant (DPPH IC50 of 55.7 μg/ml). The oil also demonstrated to possess high antibacterial activity even against multidrug resistant strains of Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli and Klebsiella pneumoniae and antifungal activity against Candida albicans and Non-Candida albicans, Candida parapsilosis and Candida tropicalis. The results of this study, in conjunction with already published data on the properties of other Cymbopogon citratus oils, provide evidence that it could have a potential application as food preservative.info:eu-repo/semantics/publishedVersio

Assessment of the plant growth promotion abilities of six bacterial isolates using Zea mays as indicator plant

Zea mays, one of the most important cereals worldwide, is a plant not only with food and energy value, but also with phytoremediation potential. The use of plant growth promoting (PGP) rhizobacteria may constitute a biological alternative to increase crop yield and plant resistance to degraded environments. In search for PGP rhizobacteria strains, 6 bacterial isolates were isolated from a metal contaminated site, screened in vitro for their PGP characteristics and their effects on the growth of Z. mays were assessed. Isolates were identified as 3A10T, ECP37T, corresponding to Chryseobacterium palustre and Chryseobacterium humi, and 1ZP4, EC15, EC30 and 1C2, corresponding to strains within the genera Sphingobacterium, Bacillus, Achromobacter, and Ralstonia, respectively. All the bacterial isolates were shown to produce indole acetic acid, hydrogen cyanide and ammonia when tested in vitro for their plant growth promoting abilities, but only isolates 1C2, 1ZP4 and ECP37T have shown siderophore production. Their further application in a greenhouse experiment using Z. mays indicated that plant traits such as root and shoot elongation and biomass production, and nutrient status, namely N and P levels, were influenced by the inoculation, with plants inoculated with 1C2 generally outperforming the other treatments. Two other bacterial isolates, 1ZP4 and ECP37T also led to increased plant growth in the greenhouse. These 3 species, corresponding to strains within the genera Ralstonia (1C2), Sphingobacterium (1ZP4), and to a strain identified as C. humi (ECP37T) can thus be potential agents to increase crop yield in maize plants.info:eu-repo/semantics/acceptedVersio

Old and new challenges in Parkinson's disease therapeutics

Parkinson's disease (PD) is a neurodegenerative disorder characterized by the degeneration of dopaminergic neurons and/or loss od neuronal projections, in several dopaminergic networks. Current treatments for idiopathic PD rely mainly on the use of pharmacologic agents to improve motor symptomatology of PD patients. Nevertheless, so far PD remains an incurable disease. Therefore, it is of utmost importance to establish new therapeutic strategies for PD treatment. Over the last 20 years, several molecular, gene and cell/stem-cell therapeutic approaches have been developed with the aim of counteracting or retarding PD progression. The scope of this review is to provide an overview of PD related therapies and major breakthroughs achieved within this field. In order to do so, this review will start by focusing on PD characterization and current treatment options covering thereafter molecular, gene and cell/stem cell-based therapies that are currently being studied in animal models of PD or have recently been tested in clinical trials. Among stem cell-based therapies, those using MSCs as possible disease modifying agents for PD therapy and, specifically, the MSCs secretome contribution to meet the clinical challenge of counteracting or retarding PD progression, will be more deeply explored.Portuguese Foundation for Science and Technology (FCT) for the PhD fellowship attributed to A.O. Pires (Reference: SFRH/BD/33900/2009) and the IF development grant to A.J. Salgado (Reference: IF/00111/2013). Project NORTE-01-0145-FEDER-000013, supported by the Northern Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (FEDER). Funded by FEDER funds, through the Competitiveness Factors Operational Programme (COMPETE), and by National funds, through the Foundation for Science and Technology (FCT), under the scope of the project POCI-01-0145-FEDER-007038info:eu-repo/semantics/publishedVersio

The secretome of bone marrow and wharton jelly derived mesenchymal stem cells induces differentiation and neurite outgrowth in Sh-SY5Y cells

The goal of this study was to determine and compare the effects of the secretome of mesenchymal stem cells (MSCs) isolated from human bone-marrow (BMSCs) and the Wharton jelly surrounding the vein and arteries of the umbilical cord (human umbilical cord perivascular cells (HUCPVCs)) on the survival and differentiation of a human neuroblastoma cell line (SH-SY5Y). For this purpose, SH-SY5Y cells were differentiated with conditioned media (CM) from the MSCs populations referred above. Retinoic acid cultured cells were used as control for neuronal differentiated SH-SY5Y cells. SH-SY5Y cells viability assessment revealed that the secretome of BMSCs and HUCPVCs, in the form of CM, was able to induce their survival. Moreover, immunocytochemical experiments showed that CM from both MSCs was capable of inducing neuronal differentiation of SH-SY5Y cells. Finally, neurite lengths assessment and quantitative real-time reverse-transcription polymerase chain reaction (RT-PCR) analysis demonstrated that CM from BMSCs and HUCPVCs differently induced neurite outgrowth and mRNA levels of neuronal markers exhibited by SH-SY5Y cells. Overall, our results show that the secretome of both BMSCs and HUCPVCs was capable of supporting SH-SY5Y cells survival and promoting their differentiation towards a neuronal phenotype.The authors would like to thank Foundation Calouste de Gulbenkian for the funds attributed to Antonio J. Salgado, Portuguese Science Foundation of Science and Technology (FCT) for the Ph.D. fellowship attributed to Ana O. Pires (reference: SFRH/BD/33900/2009), and IF Development Grant to Antonio J. Salgado. They also would like to acknowledge Professor J. E. Davies (University of Toronto, Canada) and Professor Patricia Maciel (University of Minho, Portugal) for kindly providing HUCPVCs and SH-SY5Y cells, respectively. Finally, the authors would like to acknowledge the contribution of Fabio Rodrigues, Nuno Silva, Silvina Samy, and Vera Cardoso to this work

Testosterone represses ubiquitin ligases atrogin-1 and Murf-1 expression in an androgen-sensitive rat skeletal muscle in vivo

Pires-Oliveira M, Maragno AL, Parreiras-E-Silva LT, Chiavegatti T, Gomes MD, Godinho RO. Testosterone represses ubiquitin ligases atrogin-1 and Murf-1 expression in an androgen-sensitive rat skeletal muscle in vivo. J Appl Physiol 108: 266-273, 2010. First published November 19, 2009; doi:10.1152/japplphysiol.00490.2009.-Skeletal muscle atrophy induced by denervation and metabolic diseases has been associated with increased ubiquitin ligase expression. in the present study, we evaluate the influence of androgens on muscle ubiquitin ligases atrogin-1/MAFbx/FBXO32 and Murf-1/Trim63 expression and its correlation with maintenance of muscle mass by using the testosterone-dependent fast-twitch levator ani muscle (LA) from normal or castrated adult male Wistar rats. Gene expression was determined by qRT-PCR and/or immunoblotting. Castration induced progressive loss of LA mass (30% of control, 90 days) and an exponential decrease of LA cytoplasm-to-nucleus ratio (nuclear domain; 22% of control after 60 days). Testosterone deprivation induced a 31-fold increase in LA atrogin-1 mRNA and an 18-fold increase in Murf-1 mRNA detected after 2 and 7 days of castration, respectively. Acute (24 h) testosterone administration fully repressed atrogin-1 and Murf-1 mRNA expression to control levels. Atrogin-1 protein was also increased by castration up to 170% after 30 days. Testosterone administration for 7 days restored atrogin-1 protein to control levels. in addition to the well known stimulus of protein synthesis, our results show that testosterone maintains muscle mass by repressing ubiquitin ligases, indicating that inhibition of ubiquitin-proteasome catabolic system is critical for trophic action of androgens in skeletal muscle. Besides, since neither castration nor androgen treatment had any effect on weight or ubiquitin ligases mRNA levels of extensor digitorum longus muscle, a fast-twitch muscle with low androgen sensitivity, our study shows that perineal muscle LA is a suitable in vivo model to evaluate regulation of muscle proteolysis, closely resembling human muscle responsiveness to androgens.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Dept Pharmacol, BR-04044020 São Paulo, BrazilUniv São Paulo, Fac Med Ribeirao Preto, Dept Biochem & Immunol, Ribeirao Preto, BrazilUniversidade Federal de São Paulo, Dept Pharmacol, BR-04044020 São Paulo, BrazilFAPESP: 05/59006-1FAPESP: 2006/58629-8Web of Scienc

Novel hydroxyamides and amides containing D-glucopyranose or D-fructose units: biological assays in MCF-7 and MDST8 cell lines

A novel library of 15 compounds, hydroxyamides and amides containing a β-d-glucopyranose (d-Gluc) or a β-d-fructose (d-Fruc) units was designed and synthesized for antiproliferative assays in breast (MCF-7) and colon (MDST8) cancer cell lines. Twelve of them were hydroxyamides and were successfully synthesized from β-d-glucuronic acid (d-GluA). Six of these hydroxyamides which were acetylated hydroxy-β-d-glucopyranuronamide 2a–2f (1st Family) and the other six were their respective isomers, that is, hydroxy-β-d-fructuronamide 3a–3f (2nd Family), obtained by acid–base catalyzed isomerization. These compounds have the general structure, d-Glucsingle bondCdouble bond; length as m-dashONHsingle bondCHRsingle bond(CH2)nsingle bondOH and d-Frucsingle bondCdouble bond; length as m-dashONHsingle bondCHRsingle bond(CH2)nsingle bondOH, where R = an aromatic, alkyl or a hydrogen substituent, with n = 0 or 1. Eight of these contained a chiral aminoalcohol group. Three compounds were amides containing a d-glucopyranose unit (3rd Family). SAR studies were conducted with these compounds. Antiproliferative studies showed that compound 4a, the bromo-amide containing the β-d-glucopyranose ring, potently inhibits the proliferation of the MDST8 cells. Five compounds (2e, 2f, 3d, 3e, and 3f) were shown to potently selectively inhibit the proliferation of the MCF-7 cells. Compound 4b was the only one showing inhibition in both cell lines. In general, the more active compounds were the amides and hydroxyamides containing the β-d-fructose moiety, and containing an alkyl group or hydrogen. Half-inhibitory concentrations (IC50) of between 0.01 and 10 μM, were observed

Predicting the distribution of canine leishmaniasis in western Europe based on environmental variables.

The domestic dog is the reservoir host of Leishmania infantum, the causative agent of zoonotic visceral leishmaniasis endemic in Mediterranean Europe. Targeted control requires predictive risk maps of canine leishmaniasis (CanL), which are now explored. We databased 2187 published and unpublished surveys of CanL in southern Europe. A total of 947 western surveys met inclusion criteria for analysis, including serological identification of infection (504, 369 dogs tested 1971-2006). Seroprevalence was 23 2% overall (median 10%). Logistic regression models within a GIS framework identified the main environmental predictors of CanL seroprevalence in Portugal, Spain, France and Italy, or in France alone. A 10-fold cross-validation approach determined model capacity to predict point-values of seroprevalence and the correct seroprevalence class (20%). Both the four-country and France-only models performed reasonably well for predicting correctly the 20% seroprevalence classes (AUC >0 70). However, the France-only model performed much better for France than the four-country model. The four-country model adequately predicted regions of CanL emergence in northern Italy (<5% seroprevalence). Both models poorly predicted intermediate point seroprevalences (5-20%) within regional foci, because surveys were biased towards known rural foci and Mediterranean bioclimates. Our recommendations for standardizing surveys would permit higher-resolution risk mapping

Novel Perspectives in Management of Angiogenesis and Cancer Therapy

Funding: The project was funded by IPOLFG EPE and by iNOVA4Health (UID/Multi/04462/2019) a program financially supported by Fundação para a Ciência e Tecnologia (FCT)/Ministério da Educação e Ciência, through national funds. We also acknowledge funding from FCT-MCTES through the project DREAM—PTDC/MEC-ONC/29327/2017. FL-C PhD fellowship was funded by FCT (PD/BD/128337/2017).The activation of endothelial cells (ECs) is a crucial step on the road map of tumor angiogenesis and expanding evidence indicates that a pro-oxidant tumor microenvironment, conditioned by cancer metabolic rewiring, is a relevant controller of this process. Herein, we investigated the contribution of oxidative stress-induced ferroptosis to ECs activation. Moreover, we also addressed the anti-angiogenic effect of Propranolol. We observed that a ferroptosis-like mechanism, induced by xCT inhibition with Erastin, at a non-lethal level, promoted features of ECs activation, such as proliferation, migration and vessel-like structures formation, concomitantly with the depletion of reduced glutathione (GSH) and increased levels of oxidative stress and lipid peroxides. Additionally, this ferroptosis-like mechanism promoted vascular endothelial cadherin (VE-cadherin) junctional gaps and potentiated cancer cell adhesion to ECs and transendothelial migration. Propranolol was able to revert Erastin-dependent activation of ECs and increased levels of hydrogen sulfide (H2S) underlie the mechanism of action of Propranolol. Furthermore, we tested a dual-effect therapy by promoting ECs stability with Propranolol and boosting oxidative stress to induce cancer cell death with a nanoformulation comprising selenium-containing chrysin (SeChry) encapsulated in a fourth generation polyurea dendrimer (SeChry@PUREG4). Our data showed that novel developments in cancer treatment may rely on multi-targeting strategies focusing on nanoformulations for a safer induction of cancer cell death, taking advantage of tumor vasculature stabilization.publishersversionpublishe