A novel antiviral approach.

Viral infections are often the etiological agents of severe acute and chronic human diseases. Their peculiar biology usually leads to the need of design specific therapies for each virus, and the eradication of the viruses and the healing of the patients very often are not reached also after decades of theoretical and applied researches. HIV is a classical example of how the efforts of the researchers may be disappointed in eradicating a virus infection in an infected patient. Here I present a hypothesis for a new antiviral approach that may be suitable for the treatment of HIV infected patients. The same approach, with opportune modifications, may be also applied as healing strategy for a wide set of viruses infections. In brief, my idea is to use the retrotranscription machinery and the packaging system of HIV infected cells to amplify the interfering effects of siRNAs directed against HIV genes and transcripts. The coding sequences for the interfering RNAs are brought to the infected cells via modified HIV virions deficient for structural viral genes that will use the resident viral activities of HIV infected cell as helpers. The use of this strategy will probably lead to an intracellular, intercellular and systemic amplification of the specific virus-targeted interfering activities. Moreover this strategy may show novel levels of interference: a competition between the deficient and wild type viruses for the packaging molecules and the possibility of homologous recombination between the deficient and wild type viruses that may lead in turn to the formation of recombinant non infectious viruses, and the removing of wild type provirus sequence from the host genome of infected cells by recombination

Hutchinson Gilford Progeria Syndrome: A Therapeutic Approach via Adenoviral Delivery of CRISPR/cas Genome Editing System

Hutchinson-Gilford Progeria Syndrome (HGPS) is a rare human genetic disease caused by mutations in the LMNA gene. LMNA codes for structural components of the nuclear lamina. Alterations of nuclear lamina lead to a very variable class of diseases known as laminopathies. In detail, HGPS manifests a severe premature ageing phenotype due to the accumulation of a dominant negative form of lamin-A called progerin. With current treatments, the life expectancy of HGPS patients does not exceed their second decade. Death is usually due to cardiovascular complications. Recently, a new technology for mammals in vivo gene editing has been developed: the clustered regularly interspaced short palindromic repeats/Cas protein (CRISPR/Cas) system. The CRISPR/Cas technology permits to edit the genome at specific loci. Even if the CRSIPR/Cas constructs are transiently administered to the target cells, the genome editing is permanent. The advantages of the combination of non-integrating transient vectors in combination with the CRISPR/Cas constructs could give rise to a secure approach for the treatment of disease of genetic origin, especially those caused by dominant negative mutations, such as HGPS. A potential application of non-integrating transient vectors carrying CRISPR/Cas constructs for the treatment of HGPS will be discussed in detail

A ceRNA approach may unveil unexpected contributors to deletion syndromes, the model of 5q- syndrome

In genomic deletions, gene haploinsufficiency might directly configure a specific disease phenotype. Nevertheless, in some cases no functional association can be identified between haploinsufficient genes and the deletion-associated phenotype. Transcripts can act as microRNA sponges. The reduction of transcripts from the hemizygous region may increase the availability of specific microRNAs, which in turn may exert in-trans regulation of target genes outside the deleted region, eventually contributing to the phenotype. Here we prospect a competing endogenous RNA (ceRNA) approach for the identification of candidate genes target of epigenetic regulation in deletion syndromes. As a model, we analyzed the 5q- myelodysplastic syndrome. Genes in haploinsufficiency within the common 5q deleted region in CD34+ blasts were identified in silico. Using the miRWalk 2.0 platform, we predicted microRNAs whose availability, and thus activity, could be enhanced by the deletion, and performed a genomewide analysis of the genes outside the 5q deleted region that could be targeted by the predicted miRNAs. The analysis pointed to two genes with altered expression in 5q- transcriptome, which have never been related with 5q- before. The prospected approach allows investigating the global transcriptional effect of genomic deletions, possibly prompting discovery of unsuspected contributors in the deletion-associated phenotype. Moreover, it may help in functionally characterizing previously reported unexpected interactions

RNA mediated trans-activation: its therapeutic potential in anaplastic thyroid cancer

RNA mediated trans-Activation is a proposed mechanism involved in the setting of the epigenetic state of chromatin. It has been studied first in Drosophila melanogaster where it seems that at least some transcribed loci are marked as transcriptionally active by their own transcripts. This effect acts in trans and in some cases could give rise to a transgenerational paramutational-like effect. This work studies RNA mediated trans-activation in the light of one of its possible applications, specifically its use as a therapeutic strategy for the incurable and highly aggressive ATC (anaplastic thyroid cancer). We explore the possibility to reactivate the Thyroid specific NIS (Natrium-Iodine symport) via the expression of ncRNAs with sequence homology with transcripts from the NIS coding locus itself. This work suggests that RNA trans-activation is a conserved mechanism and it may be used in human to manipulate the gene expression

Cómo realizar un plan hoshin: una aplicación. En Ssc. Telesí S.R.L.

Este trabajo es una introducción práctica al uso de la gestión hoshin en empresas pequeñas y medianas (pymes). En particular, refleja nuestra experiencia de su aplicación en Asc. Telesí, una firma de mantenimiento y venta de ascensores, con sede en Buenos Aires. Presentamos los conceptos generales de la gestión hoshin y los contrastamos con los de la administración por objetivos. Luego de discutir su aplicabilidad en las pymes, describimos la empresa, su estrategia y la forma en que desarrollamos anualmente la gestión hoshin, dentro de un ciclo de planificación de varios años. Destacamos dos enfoques ligeramente distintos empleados en Asc. Telesí en dos etapas sucesivas, como parte de un proceso de aprendizaje a través de la mejora continua. Resumimos, finalmente, un procedimiento general para la planificación hoshin en una pyme y proponemos algunas preguntas para reflexionar sobre el tema.Hoshin kanri, catchball, planificación por objetivos, planificación por políticas, pequeña y mediana empresa, metodología de planificación.

Identification of Novel Wsf1 Mutations in a Sicilian Child with Wolfram Syndrome

Wolfram Syndrome (WS) is a rare hereditary disease with autosomal recessive inheritance with incomplete penetrance. It is characterized by diabetes mellitus associated with progressive optic atrophy. The diagnosis is essentially clinical and mutation analysis is used to confirm the diagnosis. In the present study we describe the clinical and molecular features of a diabetic child carrying two novel WFS1 mutations. The Sicilian proband and his non-affected family were studied. Ophthalmologic examination included: visual acuity determination and funduscopy, optical coherent tomography, retinal fluorangiography, perimetry and electroretinogram. Molecular methods: automatic sequencing of PCR amplified WFS1 gene fragments and qRT-PCR analysis of WFS1 transcripts. 3 WSF1 mutations have been identified in the proband. One allele carries 2 paternally inherited mutations (c.1332 C>G and c.1631C>G) in exon-8, never annotated before, in heterozygosis with one “de novo” classic mutation (c.505 G>A) in exon-5. In addition, we report an unexpected molecular feature: higher WFS1 mRNA levels in the proband compared to the father

Targeted sequencing of BRAF by MinION in archival Formalin-Fixed Paraffin-Embedded specimens allows to discriminate between Hairy Cell Leukemia and Hair Cell Leukemia Variant

Targeted sequencing of BRAF by MinION in archival Formalin-Fixed Paraffin-Embedded specimens allows to discriminate between Hairy Cell Leukemia and Hair Cell Leukemia Varian

Commentary: Analysis of SUMO1-conjugation at synapses

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