Prevention of Herpesviridae Infections by Cationic PEGylated Carbosilane Dendrimers

Infections caused by viruses from the Herpesviridae family produce some of the most prevalent transmitted diseases in the world, constituting a serious global public health issue. Some of the virus properties such as latency and the appearance of resistance to antiviral treatments complicate the development of effective therapies capable of facing the infection. In this context, dendrimers present themselves as promising alternatives to current treatments. In this study, we propose the use of PEGylated cationic carbosilane dendrimers as inhibitors of herpes simplex virus 2 (HSV-2) and human cytomegalovirus (HCMV)infections. Studies of mitochondrial toxicity, membrane integrity, internalization and viral infection inhibition indicated that G2-SN15-PEG, G3-SN31-PEG, G2-SN15-PEG fluorescein isothiocyanate (FITC) labeled and G3-SN31-PEG-FITC dendrimers are valid candidates to target HSV-2 and HCMV infections since they are biocompatible, can be effectively internalized and are able to significantly inhibit both infections. Later studies (including viral inactivation, binding inhibition, heparan sulphate proteoglycans (HSPG)binding and surface plasmon resonance assays) confirmed that inhibition takes place at first infection stages. More precisely, these studies established that their attachment to cell membrane heparan sulphate proteoglycans impede the interaction between viral glycoproteins and these cell receptors, thus preventing infection. Altogether, our research confirmed the high capacity of these PEGylated carbosilane dendrimers to prevent HSV-2 and HCMV infections, making them valid candidates as antiviral agents against Herpesviridae infections

Carbohydrate effect of novel arene Ru(II) phenanthroline-glycoconjugates on metastatic biological processes

Novel water-soluble half-sandwich ruthenium(II) polypyridyl-glycoconjugates [Ru(p-cymene)Cl{N-(1,10-phenanthroline-5-yl)-& beta;-glycopyranosylamine}][Cl] (glycopyranosyl = D-glucopyranosyl (1), D-mannopyranosyl (2), L-rhamnopyranosyl (3) and L-xylopyranosyl (4)) have been synthesized and fully characterized. Their behaviour in water under physiological conditions has been studied by nuclear magnetic resonance spectroscopy, revealing their hydrolytic stability. Interactions of the novel compounds with duplex-deoxiribonucleic acid (dsDNA) were investigated by different techniques and the results indicate that, under physiological pH and saline conditions, the metal glycoconjugates bind DNA in the minor groove and/or through external, electrostatic interactions, and by a non-classical, partial intercalation mechanism in non-saline phosphate buffered solution. Effects of compounds 1-4 on cell viability have been assessed in vitro against two human cell lines (androgen-independent prostate cancer PC-3 and non-tumorigenic prostate RWPE-1), showing moderate cytotoxicities, with IC50 values higher than those found for free ligands [N-(1,10-phenanthroline-5-yl)-& beta;-glycopyranosylamine] (glycopyranosyl = D-glucopyranosyl (a), D-mannopyranosyl (b), L-rhamnopyranosyl (c) and L-xylopyranosyl (d)) or corresponding metal-aglycone. Cell viability was assayed in the presence and absence of the glucose transporters (GLUTs) inhibitor [N4-{1-(4-cyanobenzyl)-5-methyl-3-(trifluoromethyl)-1H-pyrazol-4-yl}-7-fluoroquinoline-2,4dicarboxamide] (BAY-876), and the results point to a negligible impact of the inhibition of GLUTs on the cytotoxicity caused by Ru(II) compounds 1-4. Remarkably, glycoconjugates 1-4 potently affect the migration pattern of PC-3 cells, and the wound healing assay evidence that the presence of the carbohydrate and the Ru(II) center is a requisite for the anti-migratory activity observed in these novel derivatives. In addition, derivatives 1-4 strongly affect the matrix metalloproteinase MMP-9 activities of PC-3 cells, while proMMP-2 and especially proMMP-9 were influenced to a much lesser extent

Water soluble, optically active monofunctional Pd(II) and Pt(II) compounds: promising adhesive and antimigratory effects on human prostate PC-3 cancer cells

New water soluble, enantiopure palladium and platinum compounds R-N-[M{(1S, 4R)-kappa NOH,kappa(NH)-N-2(2-pic)} Cl]Cl and S-N-[M{(1R, 4S)-kappa NOH,kappa(NH)-N-2(2-pic)}Cl]Cl (2-pic = 2-picolyl, M = Pd 1 and 1', Pt 2 and 2', respectively), and heterometallic Pd/Ti [(eta(5)-C5H5)(2)Ti{(1S, 4R)-kappa ON,kappa(NH)-N-2(2-pic)}(PdCl)] Cl (3) have been synthesized. These novel compounds were fully characterized by NMR spectroscopy and CHN elemental analysis and 1, 1', 2 and 2' were further evaluated by polarimetry, ultra-violet and circular dichroism spectroscopy. The aqueous stability of novel compounds was studied by NMR spectroscopy under physiological conditions and the new species detected under such conditions have been characterized by NMR techniques and HR-ESI-MS (High-Resolution Electrospray Ionization Mass Spectrometry). CompoundDNA interactions have been investigated for the palladium and platinum compounds by equilibrium dialysis, Fluorescence Resonance Energy Transfer (FRET) DNA melting assays and viscometric titrations, revealing a better binding affinity and ability to affect duplex DNA of the palladium compounds. Metal derivatives have been tested in vitro against three cancer (prostate PC-3, cervical HeLa and breast MCF-7) and one non-tumorigenic (human prostate RWPE-1) cell lines. The highest anticancer activities were shown by palladium compounds 1 and 1' in all cancer lines, although their toxicity was lower than that found for cisplatin. Most importantly, the effect of the compounds on the cell adhesion and migration of the androgen- independent prostate cancer PC-3 cells has been assessed, and the efficacy of Pd enantiomers to affect the invasive phenotype of PC-3 cells has been demonstrated.Universidad de AlcaláComunidad de Madri

Stereoselective synthesis of oxime containing Pd(II) compounds: Highly effective, selective and stereo-regulated cytotoxicity against carcinogenic PC-3 cells

New palladium compounds [Pd{(1S,4R)-NOHNH(R)}Cl-2] (R = Ph 1a or Bn 1b), [Pd{(1S,4R)-NOHNH(R)}{(1S,4R)-NONH(R)}][Cl] (R = Ph 2a or Bn 2b) and corresponding [Pd{(1R,4S)-NOHNH(R)}Cl-2] (R = Ph 1a' or Bn 1b') and [Pd{(1R,4S)-NOHNH(R)}{(1R,4S)-NONH(R)}][Cl] (R = Ph 2a' or Bn 2b') have been synthesized. Novel compounds 1a, 1b, and 2b (and 1a', 1b', and 2b') were obtained in solution as a mixture of diastereomers whose relative ratios depend on the solvent and the nature of the amino substituent. In contrast, the synthetic reactions of derivatives 2a and 2a' were stereospecific, and afforded single enantiomers of absolute configuration (S-N,1S(C),4R(C))-(R-N,1S(C),4R(C)) and (R-N,1R(C),4S(C))-(S-N,1R(C),4S(C)), respectively. All compounds have been fully characterized by NMR and IR spectroscopy, time-dependent UV-spectroscopy, ESI-HR-MS in water, and CHN elemental analysis. Absolute configurations of the major epimers of 1a and 1a', both epimers of 1b and enantiomer 2a', were determined by single crystal X-ray crystallography, and confirmed by 2D NOESY NMR experiments in solution. Additionally, the pH-dependent stability of 2b in water was assessed by H-1-NMR spectroscopy. Metal derivatives have been tested in vitro against three human cancer (prostate PC-3, cervical HeLa, and breast MCF-7) cell lines. The highest anticancer activities were shown by palladium compound 2a' in all cancer cells, with IC50 values up to 80 times lower than those found for cisplatin. The cytotoxicity of 2a and 2a '' is stereo-dependent, with IC50 values that differ significantly for each enantiomer in all the cell lines tested. The cytotoxic activity of 2a and 2a' was further evaluated against the non-tumorigenic human prostate RWPE-1 cell line, revealing a selectivity index (SI) of ca. 30 for derivative 2a'. DNA interactions have been investigated by equilibrium dialysis, fluorescence resonance energy transfer (FRET) DNA melting assays, and viscometric titrations, pointing to groove and/or external binding. Cell cycle assay on PC-3 cells after treatment with 2a or 2a' shows cell cycle arrest in the S and G2/M phases, especially when the cells are treated with compound 2a'.Ministerio de Ciencia e InnovaciónMinisterio de Economía, Industria y CompetitividadUniversidad de AlcaláComunidad de Madri

Effectiveness of an intervention for improving drug prescription in primary care patients with multimorbidity and polypharmacy:Study protocol of a cluster randomized clinical trial (Multi-PAP project)

This study was funded by the Fondo de Investigaciones Sanitarias ISCIII (Grant Numbers PI15/00276, PI15/00572, PI15/00996), REDISSEC (Project Numbers RD12/0001/0012, RD16/0001/0005), and the European Regional Development Fund ("A way to build Europe").Background: Multimorbidity is associated with negative effects both on people's health and on healthcare systems. A key problem linked to multimorbidity is polypharmacy, which in turn is associated with increased risk of partly preventable adverse effects, including mortality. The Ariadne principles describe a model of care based on a thorough assessment of diseases, treatments (and potential interactions), clinical status, context and preferences of patients with multimorbidity, with the aim of prioritizing and sharing realistic treatment goals that guide an individualized management. The aim of this study is to evaluate the effectiveness of a complex intervention that implements the Ariadne principles in a population of young-old patients with multimorbidity and polypharmacy. The intervention seeks to improve the appropriateness of prescribing in primary care (PC), as measured by the medication appropriateness index (MAI) score at 6 and 12months, as compared with usual care. Methods/Design: Design:pragmatic cluster randomized clinical trial. Unit of randomization: family physician (FP). Unit of analysis: patient. Scope: PC health centres in three autonomous communities: Aragon, Madrid, and Andalusia (Spain). Population: patients aged 65-74years with multimorbidity (≥3 chronic diseases) and polypharmacy (≥5 drugs prescribed in ≥3months). Sample size: n=400 (200 per study arm). Intervention: complex intervention based on the implementation of the Ariadne principles with two components: (1) FP training and (2) FP-patient interview. Outcomes: MAI score, health services use, quality of life (Euroqol 5D-5L), pharmacotherapy and adherence to treatment (Morisky-Green, Haynes-Sackett), and clinical and socio-demographic variables. Statistical analysis: primary outcome is the difference in MAI score between T0 and T1 and corresponding 95% confidence interval. Adjustment for confounding factors will be performed by multilevel analysis. All analyses will be carried out in accordance with the intention-to-treat principle. Discussion: It is essential to provide evidence concerning interventions on PC patients with polypharmacy and multimorbidity, conducted in the context of routine clinical practice, and involving young-old patients with significant potential for preventing negative health outcomes. Trial registration: Clinicaltrials.gov, NCT02866799Publisher PDFPeer reviewe

Retrospective evaluation of whole exome and genome mutation calls in 746 cancer samples

Funder: NCI U24CA211006Abstract: The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) curated consensus somatic mutation calls using whole exome sequencing (WES) and whole genome sequencing (WGS), respectively. Here, as part of the ICGC/TCGA Pan-Cancer Analysis of Whole Genomes (PCAWG) Consortium, which aggregated whole genome sequencing data from 2,658 cancers across 38 tumour types, we compare WES and WGS side-by-side from 746 TCGA samples, finding that ~80% of mutations overlap in covered exonic regions. We estimate that low variant allele fraction (VAF < 15%) and clonal heterogeneity contribute up to 68% of private WGS mutations and 71% of private WES mutations. We observe that ~30% of private WGS mutations trace to mutations identified by a single variant caller in WES consensus efforts. WGS captures both ~50% more variation in exonic regions and un-observed mutations in loci with variable GC-content. Together, our analysis highlights technological divergences between two reproducible somatic variant detection efforts

Aplicaciones terapéuticas de sistemas dendríticos frente a infecciones virales: Virus de la Inmunodeficiencia Humana Tipo 1, Virus el Herpes Simple Tipo 2 y Citomegalovirus Humano

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de Lectura: 21-10-2022Despite being the cause of epidemics for thousands of years, the study of viruses is relatively young. However, the appearance of previously unknown viruses and the spread of already known viruses in new areas have led to a massive increase in virology research. Refined approaches in the study of viruses have allowed for an in depth understanding of the molecular, evolutionary, and epidemiological aspects of viruses, which has reduced the mortality and morbidity of these infectious diseases. Nevertheless, viral infections remain one of the major global health problems, mostly in vulnerable populations such as immunocompromised patients and people from developing countries. To illustrate, the global population living with Human Immunodeficiency Virus-1 (HIV-1) exceeds 37 million people and each year there are more than 1,5 million new infections. Another sexually transmitted viral pathogen is Herpes Simplex Virus-2 (HSV-2), the causative agent of more than 490 million infections around the world. However, sexual transmission is not the only way by which viral pathogens are widely disseminated, for example Human Cytomegalovirus (HCMV), which affects more than 50% of the global population and reaches values of more than 90% in some regions, is mostly acquired during childhood or through organ transplant or blood transfusion. There are currently different therapies for diseases produced by these three viruses, but there is no definitive cure and treatments face diverse challenges such as timely diagnosis, access and adherence to treatment or appearance of resistances. In this scenario, nanotechnology presents itself as a promising tool for the development of new antiviral therapies. In this work, novel PEGylated cationic carbosilane dendrimers (PCCDs) have demonstrated to be efficient for viral inhibition in two different approaches: their use as delivery vehicles and their antiviral activity per se. In the first approach, the use as delivery systems, these dendrimers have proven to be biocompatible and effectively delivered into target cells. In addition, they have proved to form stable complexes with miRNAs that present anti-HIV-1 activity, which significantly and specifically improved the inhibition capacity of these RNA molecules by themselves. In the second approach, the antiviral activity per se, PCCDs have shown to effectively inhibit the attachment of viral glycoproteins from HSV-2 and HCMV to heparan sulphate proteoglycans, thus preventing the infection of target cells. Both therapeutic strategies have demonstrated that G2-SN15-PEG and G3-SN31-PEG dendrimers are promising candidates to be used against Retroviridae and Herpesviridae infections. Last part of this work consisted in the study of the application of micellar carbosilane dendrons in the development of a dendritic cell (DC)-based therapeutic vaccine for HIV-1. Dendrimicelles were shown to be valid carriers of HIV-1-derived peptides into moDCs, which induced them to a slight maturation. Experiments with T and B cell activation and release of inflammatory cytokines confirmed a slight stimulation of HIVspecific immune response. Collectively, these experiments confirmed that EG3SO3Na, ChG3SO3Na, EG3NMe3I, and ChG3NMe3I dendrimicelles are valid candidates to be used in the development of a therapeutic vaccine against HIV-

Creating a research network for a successful e-portfolio design and implementation

The presence of e-portfolios in educational centres, companies and administrations has emerged strongly during the last years by creating very different practices coming from different objectives and purposes. This situation has led researchers and practitioners to design and implement e-portfolios with little reference to previous knowledge of them; consequently, developments are disparate with many of the processes and dimensions used both in development and use being unnecessary complex. In order to minimize the inconveniences, unify these developmental processes and improve the results of implementation and use of e-portfolios, it seemed necessary to create a network of researchers, teachers and trainers coming from different universities and institutions of different kinds who are interested in the investigation and the practice of e-portfolios in Spain. Therefore, The Network on e-portfolio was created in 2006, funded by the Spanish Ministry of Education and led by the Universitat Oberta de Catalunya. Besides the goals associated with the creation of this network and which we wanted to share with other European researchers and experts of other continents, we will also present in this paper some data concerned with the first study carried out on the use of e-portfolios in our country that shows where we are and which trends are the most important for the near future

Creating a research network for a successful e-portfolio design and implementation

The presence of e-portfolios in educational centres, companies and administrations has emerged strongly during the last years by creating very different practices coming from different objectives and purposes. This situation has led researchers and practitioners to design and implement e-portfolios with little reference to previous knowledge of them; consequently, developments are disparate with many of the processes and dimensions used both in development and use being unnecessary complex. In order to minimize the inconveniences, unify these developmental processes and improve the results of implementation and use of e-portfolios, it seemed necessary to create a network of researchers, teachers and trainers coming from different universities and institutions of different kinds who are interested in the investigation and the practice of e-portfolios in Spain. Therefore, The Network on e-portfolio was created in 2006, funded by the Spanish Ministry of Education and led by the Universitat Oberta de Catalunya. Besides the goals associated with the creation of this network and which we wanted to share with other European researchers and experts of other continents, we will also present in this paper some data concerned with the first study carried out on the use of e-portfolios in our country that shows where we are and which trends are the most important for the near future

Fluorescent Vitamin B₁₂–Platinum(II) Derivatives as Potential Metallotheranostic Agents for the Treatment and Imaging of Tumors

Vitamin B12 (cyanocobalamin) is an essential nutrient with very low bioavailability. Compared with normal cells, tumor cells show an increased demand for vitamin B12 to support their abnormal proliferation, which is a feature that can be exploited for the tumor-specific delivery of therapeutic and/or diagnostic agents by functionalizing vitamin B12 with suitable metallodrugs and/or luminescent probes. In this context, we report on the design of fluorescent vitamin B12–metal conjugates of the type [FLUO–B12–{M}] in which cyanocobalamin is functionalized at the 5′-site of the ribose unit with a fluorophore (FLUO: rhodamine 6G), whereas the Co(III)–cyano moiety is N-coordinated to a metal-based anticancer scaffold ({M}: Pt(II) substrate bearing enantiopure phenylamino-oxime ligands derived from R- or S-limonene). Two novel fluorescent cyanocobalamin–platinum(II) derivatives and their corresponding non-fluorescent counterparts were successfully generated and fully characterized, including the evaluation of their lipophilicity and luminescent properties. Although they exhibit low antiproliferative activity (IC50 = 40–70 μM), both fluorescent vitamin B12–platinum(II) conjugates showed an enhanced capability to inhibit cell viability compared with the inactive metal precursors and the non-fluorescent vitamin B12–platinum(II) analogues, confirming the beneficial effect of functionalization with the rhodamine 6G scaffold not only for imaging purposes but also with the aim of improving their biological activity