Adenocarcinoma de la próstata Gleason 6

This is a 69 year old patient with a medical history of ischemic heart disease, hypertension and type 2 diabetes mellitus who was admitted twice in the "Arnaldo Milián Castro" Surgical Hospital of the City of Santa Clara, Province Villa Clara, by decay and weight loss, abdominal tumors of four and 5cm respectively on the right flank and left hemiparesis with neurological symptoms, physical examination was found to further enlargement of the thyroid. He had a long evolution in living and he performed various add -abdominal and thyroid ultrasound, chest  X-ray and computed tomography of the skul, which showed an increase in nodular thyroid volume and a parahilum left pulmonary inflammatory process, not reached a definitive clinical diagnosis and died 55 days after admission.Se trata de un paciente de 69 años de edad con antecedentes personales patológicos de cardiopatía isquémica, hipertensión arterial y diabetes mellitus tipo 2 que fue ingresado en dos oportunidades en el Hospital Clínico Quirúrgico “Arnaldo Milián Castro” de la Ciudad de Santa Clara, Provincia de Villa Clara, por decaimiento y pérdida de peso, tumoraciones abdominales de cuatro y 5cm respectivamente en el flanco derecho y cuadro neurológico con hemiparesia izquierda; se constató al examen físico, además, aumento de volumen de la tiroides. Tuvo una evolución prolongada en sala y se le realizaron diversos complementarios -ultrasonido abdominal y de tiroides, rayos X de tórax y tomografía axial computadorizada de cráneo- que arrojaron un aumento de volumen nodular de la tiroides y un proceso inflamatorio pulmonar izquierdo parahiliar; no se llegó a un diagnóstico clínico definitivo y falleció 55 días después de su ingreso.

The Spectrum of FANCM Protein Truncating Variants in European Breast Cancer Cases

Germline protein truncating variants (PTVs) in the FANCM gene have been associated with a 2–4-fold increased breast cancer risk in case-control studies conducted in different European populations. However, the distribution and the frequency of FANCM PTVs in Europe have never been investigated. In the present study, we collected the data of 114 European female breast cancer cases with FANCM PTVs ascertained in 20 centers from 13 European countries. We identified 27 different FANCM PTVs. The p.Gln1701* PTV is the most common PTV in Northern Europe with a maximum frequency in Finland and a lower relative frequency in Southern Europe. On the contrary, p.Arg1931* seems to be the most common PTV in Southern Europe. We also showed that p.Arg658*, the third most common PTV, is more frequent in Central Europe, and p.Gln498Thrfs*7 is probably a founder variant from Lithuania. Of the 23 rare or unique FANCM PTVs, 15 have not been previously reported. We provide here the initial spectrum of FANCM PTVs in European breast cancer cases

An original phylogenetic approach identified mitochondrial haplogroup T1a1 as inversely associated with breast cancer risk in BRCA2 mutation carriers

Introduction: Individuals carrying pathogenic mutations in the BRCA1 and BRCA2 genes have a high lifetime risk of breast cancer. BRCA1 and BRCA2 are involved in DNA double-strand break repair, DNA alterations that can be caused by exposure to reactive oxygen species, a main source of which are mitochondria. Mitochondrial genome variations affect electron transport chain efficiency and reactive oxygen species production. Individuals with different mitochondrial haplogroups differ in their metabolism and sensitivity to oxidative stress. Variability in mitochondrial genetic background can alter reactive oxygen species production, leading to cancer risk. In the present study, we tested the hypothesis that mitochondrial haplogroups modify breast cancer risk in BRCA1/2 mutation carriers. Methods: We genotyped 22,214 (11,421 affected, 10,793 unaffected) mutation carriers belonging to the Consortium of Investigators of Modifiers of BRCA1/2 for 129 mitochondrial polymorphisms using the iCOGS array. Haplogroup inference and association detection were performed using a phylogenetic approach. ALTree was applied to explore the reference mitochondrial evolutionary tree and detect subclades enriched in affected or unaffected individuals. Results: We discovered that subclade T1a1 was depleted in affected BRCA2 mutation carriers compared with the rest of clade T (hazard ratio (HR) = 0.55; 95% confidence interval (CI), 0.34 to 0.88; P = 0.01). Compared with the most frequent haplogroup in the general population (that is, H and T clades), the T1a1 haplogroup has a HR of 0.62 (95% CI, 0.40 to 0.95; P = 0.03). We also identified three potential susceptibility loci, including G13708A/rs28359178, which has demonstrated an inverse association with familial breast cancer risk. Conclusions: This study illustrates how original approaches such as the phylogeny-based method we used can empower classical molecular epidemiological studies aimed at identifying association or risk modification effects.Peer reviewe

Analysis of mRNA abundance and stability by ribonuclease protection assay

Gene expression is a multi-step process, which proceeds from DNA through RNA to protein. The tight regulation of this process is essential for overall cellular integrity and physiological homeostasis. Regulation of the messenger RNA (mRNA) levels has emerged as a crucial event in the modulation of the expression of genetic information. The mechanisms by which this process occurs have been extensively studied and begin to be much better understood. They involve a network of complex pathways that use intrinsic features of the target mRNA, like stability, to control its relative abundance in the cytoplasm. Thus, the analysis of the mRNA stability and abundance is essential to properly undertake gene expression studies. This chapter describes the ribonuclease protection assay, a widely accepted approach to evaluate the quality and amount of a target mRNA. This technique displays a higher sensitivity than classical Northern blot analysis and may be used either individually or in combination with other quantitative methods, such as quantitative reverse-transcription PCR, as complementary procedures rendering more complete and reliable information on gene expression.Peer Reviewe

The chaperone-like activity of conserved RNA elements in the hepatitis C virus regulates genome dimerization

The RNA genome of the hepatitis C virus (HCV) establishes a network of long-distance RNA-RNA interactions that direct the progression of the infective cycle. This work shows that the dimerization of the viral genome, which is initiated at the dimer linkage sequence (DLS) within the 3′UTR, is promoted by the CRE region, while the IRES is a negative regulatory partner. Using differential 2′-acylation probing (SHAPE-dif) and molecular interference (HMX) technologies, the CRE activity was found to mainly lie in the critical 5BSL3.2 domain, while the IRES-mediated effect is dependent upon conserved residues within the essential structural elements JIIIabc, JIIIef and PK2. These findings support the idea that, along with the DLS motif, the IRES and CRE are needed to control HCV genome dimerization. They also provide evidences of a novel function for these elements as chaperone-like partners that finetune the architecture of distant RNA domains within the HCV genome.Peer reviewe

End-to-end crosstalk within the hepatitis C virus genome mediates the conformational switch of the 3′X-tail region

[EN]The hepatitis C virus (HCV) RNA genome contains multiple structurally conserved domains that make long-distance RNA–RNA contacts important in the establishment of viral infection. Microarray antisense oligonucelotide assays, improved dimethyl sulfate probing methods and 2′ acylation chemistry (selective 2’-hydroxyl acylation and primer extension, SHAPE) showed the folding of the genomic RNA 3′ end to be regulated by the internal ribosome entry site (IRES) element via direct RNA–RNA interactions. The essential cis-acting replicating element (CRE) and the 3′X-tail region adopted different 3D conformations in the presence and absence of the genomic RNA 5′ terminus. Further, the structural transition in the 3′X-tail from the replication-competent conformer (consisting of three stem-loops) to the dimerizable form (with two stem-loops), was found to depend on the presence of both the IRES and the CRE elements. Complex interplay between the IRES, the CRE and the 3′X-tail region would therefore appear to occur. The preservation of this RNA–RNA interacting network, and the maintenance of the proper balance between different contacts, may play a crucial role in the switch between different steps of the HCV cycle.Peer reviewe

The folding of the hepatitis C virus internal ribosome entry site depends on the 3′-end of the viral genome

Spanish Ministerio de Ciencia e Innovación, MICINN [BFU2009-08137 to A.B.-H., EUI2008-00158 and BIO2010-20696 to C.B.]; Junta de Andalucía [CTS-5077 to A.B.-H.]; University of Granada-GREIB Project [GREIB.PYR_2010_12 to C.R.-L.]; Spanish Council for Scientific Research [201120E004 to A.B.-H.]; FEDER funds from the EU [to A.B.-H. and C.B.]; Instituto de Salud Carlos III [to CIBERehd]. Funding for open access charge: Spanish Ministerio de Ciencia e Innovación, MICINN [BFU2009-08137 to A.B.-H.].Peer reviewe

Secuencia de RNA, construcción de RNA y DNA, y composición farmacéutica inhibidoras de la proliferación del virus causante de la hepatitis tipo C (VHC), y sus aplicaciones

Fecha de presentación internacional: 23.03.2005. - Titular: Consejo Superior de Investigaciones Científicas (CSIC)[EN] The invention relates to: an RNA sequence that inhibits the replication of the HCV virus by binding to the IRES region, RNA and DNA constructions, pharmaceutical compositions containing same, and the application thereof in methods for the prevention and treatment of infections caused, in particular, by the HCV. The aforementioned RNA sequences are advantageous in that it is a product that is synthesised naturally by cells and, as such, does not produce any type of toxic effect. In addition, the fact that the IRES region is highly conserved between the different HCV strains makes possible the development of more effective therapeutic tools against said viruses which have a good mutagenesis capacity, thereby enabling same to evade antiviral treatments.[ES] La invención proporciona una secuencia de RNA inhibidora de la replicación del virus VHC por su unión a la región IRES, así como construcciones de RNA y DNA, composiciones farmacéuticas que las contienen, y su aplicación en procedimientos de prevención y tratamiento de infecciones producidas, preferentemente por el VHC. La ventaja de dichas secuencias de RNA es que es un producto que sintetizan las células de forma natural, por lo que no se prevé ningún tipo de efecto tóxico. Además, el hecho que esta región IRES esté altamente conservada entre las distintas cepas de VHC permitirá el desarrollo de herramientas terapéuticas más eficaces frente a estos virus que presentan una gran capacidad de mutagénesis, que les capacita el escape a tratamientos antivirales.Peer reviewe

Inhibition of HCV replication and IRES-dependent translation by an RNA molecule

Hepatitis C virus (HCV) protein synthesis is mediated by a highly conserved internal ribosome entry site (IRES), mostly located at the 5’ untranslatable region (5’UTR) of the viral genome. The translation mechanism is different from that used by cellular cap-mRNAs, making IRES an attractive target site for new antiviral drugs. The present work characterizes a chimeric RNA molecule (HH363-50) composed of two inhibitors: a hammerhead ribozyme targeting position 363 of the HCV genome, and an aptamer directed towards the essential stem-loop structure in domain IV of the IRES region (which contains the translation start codon). The inhibitor RNA interferes with the formation of a translationally active complex, stalling its progression at the level of 80S particle formation. This action is likely related to the effective and specific blocking of HCV IRES-dependent translation achieved in Huh-7 cells. The inhibitor HH363-50 also reduces HCV RNA levels in a subgenomic replicon system. The present findings suggest that HH363-50 could be an effective anti-HCV compound and highlight the possibilities of antiviral agents based on RNA molecules.This work was supported by grants BFU2006-02568 from the Spanish Ministerio de Educación y Ciencia and CTS-233 from the Junta de Andalucía to A. B-H. C. R-L was funded by grant BMC2003-669. R. D-G was the recipient of a fellowship from the Spanish Ministerio de Educación y CienciaN