3-Deazaneplanocin A (DZNep), an Inhibitor of the Histone Methyltransferase EZH2, Induces Apoptosis and Reduces Cell Migration in Chondrosarcoma Cells

ObjectiveGrowing evidences indicate that the histone methyltransferase EZH2 (enhancer of zeste homolog 2) may be an appropriate therapeutic target in some tumors. Indeed, a high expression of EZH2 is correlated with poor prognosis and metastasis in many cancers. In addition, 3-Deazaneplanocin A (DZNep), an S-adenosyl-L homocysteine hydrolase inhibitor which induces EZH2 protein depletion, leads to cell death in several cancers and tumors. The aim of this study was to determine whether an epigenetic therapy targeting EZH2 with DZNep may be also efficient to treat chondrosarcomas.MethodsEZH2 expression was determined by immunohistochemistry and western-blot. Chondrosarcoma cell line CH2879 was cultured in the presence of DZNep, and its growth and survival were evaluated by counting adherent cells periodically. Apoptosis was assayed by cell cycle analysis, Apo2.7 expression using flow cytometry, and by PARP cleavage using western-blot. Cell migration was assessed by wound healing assay.ResultsChondrosarcomas (at least with high grade) highly express EZH2, at contrary to enchondromas or chondrocytes. In vitro, DZNep inhibits EZH2 protein expression, and subsequently reduces the trimethylation of lysine 27 on histone H3 (H3K27me3). Interestingly, DZNep induces cell death of chondrosarcoma cell lines by apoptosis, while it slightly reduces growth of normal chondrocytes. In addition, DZNep reduces cell migration.ConclusionThese results indicate that an epigenetic therapy that pharmacologically targets EZH2 via DZNep may constitute a novel approach to treat chondrosarcomas

PHYTOCHEMICAL PROFILING, HPTLC FINGERPRINT AND ANTIBACTERIAL, ANTI-FUNGAL, AND ANTIOXIDANT PROPERTIES OF ESSENTIAL OILS EXTRACTED FROM CUMMINUM CYMINUM, ZINGIBER OFFICINALE, TRACHYSPERMUM AMMI, ALIPNIA GALANGA, CEDRUS DEODARA, AND ELETTARIA CARDAMOMUM

Objective: The objective of the study was to carry out the phytochemical profiling of essential oils (EOs) and evaluation of their anti-microbial activity. Methods: The EOs extracted from Cumminum cyminum, Zingiber officinale, Trachyspermum ammi, Alipnia galanga, Cedrus deodara, and Elettaria cardamomum using clavenger apparatus. Phytochemical analysis and high-performance thin layer chromatography (HPTLC) fingerprinting were carried out for the EO. The antibacterial and antifungal activity were evaluated using agar well-diffusion method against two bacterial strains, Escherichia coli, Staphylococcus aureus and two fungal strains, Candida albicans, and Aspergillus brasiliensis. Positive controls ciprofloxacin-30 mg, azithromycin-15 mg, and nystatin NS-50 mg were used. Antioxidant potential of the EOs was investigated by TLC-bioautography method using 1,1-diphenyl-2-picrylhydrazyl derivatization. Results: The phytochemical analysis reveals presence of various phytochemical such as steroids, terpenoids, and phenylpropanoids. The HPTLC fingerprint is found to be unique for each of the oil. The EO of Z. officinale and T. ammi showed strong antibacterial activity against S. aureus. The EOs of C. cyminum, Tachyspermum ommi and A. galanga displayed prominent antioxidant activity on TLC bioautography. The herbs Cuminum cynimun, T. ammi, C. deodara, and Ellateria cardamomum produce reasonable amount of essentials oil, which can be explored for useful their industrial applications. Conclusions: These EOs can be explored further for their antimicrobial activity. The HPTLC analysis along with derivatization with suitable chromogenic reagents can be a rapid and simple tool for quality control of various EOs

Clostridium difficile infection.

Infection of the colon with the Gram-positive bacterium Clostridium difficile is potentially life threatening, especially in elderly people and in patients who have dysbiosis of the gut microbiota following antimicrobial drug exposure. C. difficile is the leading cause of health-care-associated infective diarrhoea. The life cycle of C. difficile is influenced by antimicrobial agents, the host immune system, and the host microbiota and its associated metabolites. The primary mediators of inflammation in C. difficile infection (CDI) are large clostridial toxins, toxin A (TcdA) and toxin B (TcdB), and, in some bacterial strains, the binary toxin CDT. The toxins trigger a complex cascade of host cellular responses to cause diarrhoea, inflammation and tissue necrosis - the major symptoms of CDI. The factors responsible for the epidemic of some C. difficile strains are poorly understood. Recurrent infections are common and can be debilitating. Toxin detection for diagnosis is important for accurate epidemiological study, and for optimal management and prevention strategies. Infections are commonly treated with specific antimicrobial agents, but faecal microbiota transplants have shown promise for recurrent infections. Future biotherapies for C. difficile infections are likely to involve defined combinations of key gut microbiota

Rosiglitazone synergizes anticancer activity of cisplatin and reduces its nephrotoxicity in 7, 12-dimethyl benz{a}anthracene (DMBA) induced breast cancer rats

<p>Abstract</p> <p>Background</p> <p>Antineoplastic drug cisplatin remains the drug of choice for various solid tumours including breast cancer. But dose dependent nephrotoxicity is the major drawback in majority of platinum based chemotherapy regimens. Recent reports have shown that inflammatory pathways are the main offender for cisplatin induced nephrotoxicity. The present study was undertaken to assess the effect of rosiglitazone, a PPARγ agonist and an anti-inflammatory agent, on cisplatin induced nephrotoxicity, and its anticancer activity in DMBA induced breast cancer rats.</p> <p>Methods</p> <p>Mammary tumours were induced in female Sprague-Dawley rats by feeding orally with dimethylbenz [a]anthracene (DMBA) (60 mg/kg). Cisplatin induced nephropathy was assessed by measurements of blood urea nitrogen, albumin and creatinine levels. Posttranslational modifications of histone H3, mitogen-activated protein (MAP) kinase p38 expression and PPAR-γ expression were examined by western blotting.</p> <p>Results</p> <p>Our data shows involvement of TNF-α in preventing cisplatin induced nephrotoxicity by rosiglitazone. Rosiglitazone pre-treatment to cisplatin increases the expression of p38, PPAR-γ in mammary tumours and shows maximum tumour reduction. Furthermore, cisplatin induced changes in histone acetylation, phosphorylation and methylation of histone H3 in mammary tumours was ameliorated by pre-treatment of rosiglitazone. Suggesting, PPAR-γ directly or indirectly alters aberrant gene expression in mammary tumours by changing histone modifications.</p> <p>Conclusion</p> <p>To best of our knowledge this is the first report which shows that pre-treatment of rosiglitazone synergizes the anticancer activity of cisplatin and minimizes cisplatin induced nephrotoxicity in DMBA induced breast cancer.</p

PMeS: Prediction of Methylation Sites Based on Enhanced Feature Encoding Scheme

Protein methylation is predominantly found on lysine and arginine residues, and carries many important biological functions, including gene regulation and signal transduction. Given their important involvement in gene expression, protein methylation and their regulatory enzymes are implicated in a variety of human disease states such as cancer, coronary heart disease and neurodegenerative disorders. Thus, identification of methylation sites can be very helpful for the drug designs of various related diseases. In this study, we developed a method called PMeS to improve the prediction of protein methylation sites based on an enhanced feature encoding scheme and support vector machine. The enhanced feature encoding scheme was composed of the sparse property coding, normalized van der Waals volume, position weight amino acid composition and accessible surface area. The PMeS achieved a promising performance with a sensitivity of 92.45%, a specificity of 93.18%, an accuracy of 92.82% and a Matthew’s correlation coefficient of 85.69% for arginine as well as a sensitivity of 84.38%, a specificity of 93.94%, an accuracy of 89.16% and a Matthew’s correlation coefficient of 78.68% for lysine in 10-fold cross validation. Compared with other existing methods, the PMeS provides better predictive performance and greater robustness. It can be anticipated that the PMeS might be useful to guide future experiments needed to identify potential methylation sites in proteins of interest. The online service is available at http://bioinfo.ncu.edu.cn/inquiries_PMeS.aspx

Water uptake by Annona diversifolia Saff. and A. purpurea Moc. & sessé ex dunal seeds (Annonaceae)

Annonaceae seeds are known by presenting dormancy mechanisms, whose reports ranging from coating impermeable to the physiological dormancy. By this way, the present study aimed to evaluate water uptake in Annona diversifolia Saff and Annona purpurea Moc & Sessé ex Dunal seeds. For this study, seeds were placed under immersion in distilled water, and used four replicates of 25 seeds of each species, which were weighed during the 480 hours that were immersed. To determine the place of purchase of water, Annona diversifolia seeds were sealed with paraffin at different locations. Based on the results, seeds from both species reached the phases I and II of water uptake, which indicates they are not hard; however, germination (Phase III) was not reached. Annona diversifolia seeds completed Phase I with, 50h and Annona purpurea with 70h from imbibitions begin, which shows that even slowly, water is acquire.As sementes de espécies da família Annonaceae são conhecidas por apresentarem mecanismos de dormência, cujos relatos vão desde a impermeabilidade do tegumento até a dormência fisiológica. Neste sentido, o objetivo do presente trabalho foi avaliar a impermeabilidade à água das sementes de Annona diversifolia Saff e Annona purpurea Moc & Sessé ex Dunal a partir do estudo da curva de aquisição de água. Para a realização do estudo, sementes foram colocadas sob imersão em água destilada, sendo utilizadas 4 repetições de 25 sementes de cada espécie, as quais foram pesadas durante as 480 horas em que ficaram imersas. Para determinar o local de aquisição de água, sementes de Annona diversifolia foram vedadas em diferentes locais com parafina. Com base nos resultados, pode-se afirmar que sementes de ambas as espécies apresentam Fase I e II da germinação, o que indica que as sementes não são impermeáveis; contudo, a Fase III não é alcançada. As sementes de Annona diversifolia completam a Fase I com 50 horas, e as de Annona purpurea, com 70 horas do início da embebição, o que demonstra que, mesmo de maneira lenta, a água é adquirida.UNESP IBB Depto BotânicaUniversidad de Ciencias y Artes de Chiapas Faculdade de Ciências BiológicasUNICACH Faculdade de Ciências BiológicasUNESP IBB Depto Botânic

Alterações nas reservas de sementes de Dalbergia nigra ((Vell.) Fr. All. ex Benth.) durante a hidratação

Seed imbibitions is the first stage of the germination process and is characterized by the hydration of tissues and cells and the activation and/or induction of the enzymes responsible for mobilizing reserves for respiration and the construction of new cell structures. The objective of this study was to investigate the alterations in reserve substances during slow hydration of Bahia Rosewood (Dalbergia nigra) seeds in water. Seeds from two different lots (Lot I and II) were placed in saturated desiccators (95-99% RH) to hydrate at 15 and 25 °C until water contents of 10, 15, 20 and 25% were reached. At each level of hydration, changes in lipid reserves, soluble carbohydrates, starch and soluble proteins were evaluated. The mobilization of reserves was similarly assessed in both lots, with no differences being observed between the two hydration temperatures. Lipid contents showed little variation during hydration, while the contents of soluble carbohydrates and starch decreased after the 15% water content level. Soluble proteins showed a gradual tendency to decrease between the control (dry seeds) up to 25% water content