Corilagin Induces High Levels of Apoptosis in the Temozolomide-Resistant T98G Glioma Cell Line

Glioblastoma multiforme (GBM), a malignant tumor of the central nervous system, has a high mortality rate; no curativetreatment is presently available and the most commonly used chemiotherapeutic drug, the alkylating agent temozolomide (TMZ), is only able to increase life expectancy and is often associated with drugresistance. Therefore, an urgent need does exist for novel drugs aimed at treating gliomas. In the present study we obtained three major results using corliagin: (a) demonstrate that it inhibits the growth of U251 glioma cells through activation of the apoptotic pathway; (b) demonstrate that it is also active on temozolomideresistant T98G glioma cells; (c) demonstrate that when used in combination with temozolomide on T98G glioma cells a higher level of pro-apototic and antiproliferative effects are observed. Our study indicates that corilagin should be investigated in more detail in order to determine if it can be developed as a potential therapeutic agent. In addition, our results suggest that corilagin could be used in combination with low dosages of other standard anticancer chemotherapeutic drugs against gliomas (such as temozolomide) with the aim of obtaining enhanced anticancer effects

Thermodynamics of Electrolytes on Anisotropic Lattices

The phase behavior of ionic fluids on simple cubic and tetragonal (anisotropic) lattices has been studied by grand canonical Monte Carlo simulations. Systems with both the true lattice Coulombic potential and continuous-space $1/r$ electrostatic interactions have been investigated. At all degrees of anisotropy, only coexistence between a disordered low-density phase and an ordered high-density phase with the structure similar to ionic crystal was found, in contrast to recent theoretical predictions. Tricritical parameters were determined to be monotonously increasing functions of anisotropy parameters which is consistent with theoretical calculations based on the Debye-H\"uckel approach. At large anisotropies a two-dimensional-like behavior is observed, from which we estimated the dimensionless tricritical temperature and density for the two-dimensional square lattice electrolyte to be $T^*_{tri}=0.14$ and $\rho^*_{tri} = 0.70$.Comment: submitted to PR

Targeting pre-miRNA by Peptide Nucleic Acids: A new strategy to interfere in the miRNA maturation

PNA conjugated to carrier peptides have been employed for the targeting of miRNA precursor, with the aim to develop molecules able to interfere in the pre-miRNA processing. The capability of the molecules to bind pre-miRNA has been tested in vitro by fluorescence assayes on Thiazole Orange labeled molecules and in vivo, in K562 cells, evaluating the amount of miRNA produced after treatment of cells with two amounts of PNAs

Preparation of anti-miR PNAs for drug development and nanomedicine

Peptide Nucleic Acids (PNAs) are oligonucleotide mimics that can be used to block the biological action of microRNA, thus affecting gene expression post-transcriptionally. PNAs are obtained with solid-phase peptide synthesis, and can be easily conjugated to other peptides. Conjugation with R8-Peptide or modification of the PNA backbone (at C5 or C2 carbon) with arginine side chains allows efficient cellular uptake. The present protocol describes the synthesis of cationic PNAs that can be used alone as drugs or for efficient co-delivery in suitable inorganic nanocarriers

Molecular methods for validation of the biological activity of peptide nucleic acids targeting microRNAs.

The involvement of microRNAs in human pathologies is a firmly established fact. Accordingly, the pharmacological modulation of their activity appears to be a very appealing issue in the development of new types of drugs (miRNA therapeutics). One of the most interesting issues is the possible development of miRNA therapeutics for development of anti-cancer molecules. In this respect appealing molecules are based on peptide nucleic acids (PNAs), displaying a pseudo-peptide backbone composed of N-(2-aminoethyl)glycine units and found to be excellent candidates for antisense and antigene therapies. The major limit in the use of PNAs for alteration of gene expression is the low uptake by eukaryotic cells. The aim of this chapter is to describe methods for determining the activity of PNAs designed to target oncomiRNAs, using as model system miR-221 and its target p27(Kip1) mRNA. The effects of PNAs targeting miR-221 are here presented discussing data obtained using as model system the human breast cancer cell line MDA-MB-231, in which miR-221 is up-regulated and p27(Kip1) down-regulated

THALAssaemia MOdular Stratification System for personalized therapy of beta-thalassemia

THALAMOSS is aimed at development of universal sets of markers and techniques for stratification of β-thalassaemia patients into treatment subgroups for (a) onset and frequency of blood transfusions, (b) choice of iron chelation, (c) induction of fetal hemoglobin, (d) prospective efficacy of gene-therapy. At present, no framework exists to guide therapeutic decisions and personalised treatment of β-thalassaemia. THALAMOSS Workpackages: WP1. Recruitment, patient characterization and development of erythroid precursor cells cultures; WP2. Omics analyses; WP3. Novel therapeutic approaches; WP4. Data analysis; WP5. Dissemination and exploitation; WP6. Regulatory and ethical issues; WP7. Management. The impact of THALAMOSS is the provision of novel biomarkers for distinct treatment subgroups in β-thalassaemia (500-1000 samples from four European medical centres), identified by combined genomics, proteomics, transcriptomics and tissue culture assays, and establishment of routine techniques for detection of these markers. Translation of these activities into the product portfolio and R&D methodology of participating SMEs will be a major issue. THALAMOSS tools and technologies will (a) facilitate identification of novel diagnostic tests, drugs and treatments specific to patient subgroups and (b) guide conventional and novel therapeutical approaches for β-thalassaemia, including personalised medical treatments. Key researchers of THALAMOSS are R.Gambari (Ferrara University, Italy), M. Kleanthous (The Cyprus Foundation for Muscular Dystrophy Research, Cyprus), S.Philipsen (Erasmus Universitair Medisch Centrum Rotterdam, The Netherlands), E.Katsantoni (Biomedical Research Foundation, Academy of Athens, Greece), S.Rivella (Weill Cornell Medical College, NY, USA), P.Holub (Masaryk University, Czech Republic), R.Galanello (Cagliari University, Italy), SL.Thein (King’s College Hospital, UK), E.Voskaridou (Laiko General Hospital, Greece). Participating SMEs are Biocep (Israel), NovaMechanics Ltd. (Cyprus) and IRBM (Italy). Industrial activities are also provided by Harbour Antibodies (The Netherlands)

Targeting the miR-221/miR-222 cluster in cancer therapy

Section of Biochemistry and Molecutal Biology, Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy MicroRNAs (miRNAs, miRs) are a family of small non-coding RNAs of 19-24 nucleotides, highly conserved during evolution, that are able to regulate gene expression by a sequence-selective targeting of mRNAs: depending on the degree of complementarity with target mRNA sequence, miRs can lead to a translational repression or mRNA degradation. MiRNAs play an essential role in malignancy. There is a large body of evidence that dysregulation of miRNAs is an hallmark of cancer; its occurs because many miRs are located in “fragile sites”, which are frequently deleted in cancer. MicroRNAs that are frequently over-expressed in neoplastic tissues compared to normal tissues and regulate tumor suppressor proteins are defined “oncomiRs”. Silencing oncomiRs could represent a novel anti-tumor approach for integrated cancer therapy. MiR-221 and miR-222 are two highly homologous miRs, whose up-regulation has been recently described in several types of human tumors. One of the main targets of miR-221/222 is the onco-suppressor p27Kip1 (CDKN1B), a gatekeepers of the G1/S cell cycle check-point CDK inhibitor protein. MiR-221/222 target the p27Kip1 mRNA, causing a down-regulation of the protein; lower levels of p27Kip1 protein lead to a state of uncontrolled proliferation, typical of aggressive tumors. In this review, we describe the role of miR-221/222 in cancer progression, focusing on the potential use as therapeutic targets for cancer

Targeting pre-miRNA by Peptide Nucleic Acids: A new strategy to interfere in the miRNA maturation.

PNAs conjugated to carrier peptides have been employed for the targeting of miRNA precursor, with the aim to develop molecules able to interfere in the pre-miRNA processing. The capability of the molecules to bind pre-miRNA has been tested in vitro by fluorescence assayes on Thiazole Orange labeled molecules and in vivo, in K562 cells, evaluating the amount of miRNA produced after treatment of cells with two amounts of PNAs