Tratamiento de imágenes de mineral acarreado post-voladura. Análisis de curvas granulométricas y comparativa mediante sistemasplit on-line y Split desktop: automático y manual

En este proyecto se han analizado distintas imágenes de fragmentos de rocas de distintas granulometrías correspondientes a una serie de voladuras de una misma cantera. Cada una de las voladuras se componen de 20 imágenes. A posteriori utilizando el programa Split Desktop en su versión 3.1, se delimitaron los fragmentos de roca de los que está compuesta la imagen, obteniéndose posteriormente la curva granulométrica correspondiente a dicha imagen. Una vez se calculan las curvas granulométricas correspondientes a cada imagen, se calcula la curva media de todas ellas, pudiéndose considerar por tanto la curva media de cada voladura. Se han utilizado las distintas soluciones del software, manual, online y automático, para realizar los análisis de dichas imágenes y a posteriori comparar sus resultados. Dichos resultados se muestran a través de una serie de gráficos y tablas que se explican con detalle para la comprensión del estudio. De dichos resultados es posible afirmar que, el tratamiento de imágenes realizado de manera online y automático por Split, desemboca en el mismo resultado, al no haber una diferencia estadística significativa. Por el contrario, el sistema manual es diferente de los otros dos, no pudiéndose afirmar cual es mejor de los dos. El manual depende del operario que trabaje las imágenes y el online de los ajustes realizados y por tanto, ambos tienen ciertas incertidumbres difíciles de solucionar. Abstract In this project, different images of rock fragments of different grain sizes corresponding to a series of blasts from the same quarry have been analyzed. To study each blast, 20 images has been used and studied with the software Split Desktop 3.1. Rock fragments from each image has been delimitated with the software, obtaining a grading curve of each one. Once these curves are calculated, the mean curve of these data set is obtained and can be considered the mean curve of each blast. Different software solutions as manual, online and automatic, has been used for the analysis of these images. Then the results has been compared between them. These results are shown through a series of graphs and tables, that are explained in detail, to enhance the understanding of the study. From these results, it can be said that the image processing with online and automatic options from Split, leads to the same result, after an statistical study. On the contrary, the manual Split mode is different from the others; however is not possible to assert what will be the best. The manual Split mode depends on the operator ability and dedication, although the online mode depends on the software settings, so therefore, both have some uncertainties that are difficult to solve.

Evaluación del funcionamiento de sistemas de análisis digital de imágenes - wipfrag (edición manual) y split online

En minería, la estimación de la curva granulométrica del escombro de voladura es importante para evaluar el diseño, ejecución y optimización de la misma. Para ello, actualmente se usan sistemas digitales de fotografías que obtienen dicha curva a partir de imágenes tomadas por una cámara. En este proyecto se ha analizado la fragmentación de seis voladuras realizadas en el año 2012 en la cantera “El Aljibe” situada en el término municipal de Almonacid de Toledo con un sistema automático en línea (Split Online) y con un software de otra compañía (WipFrag) que permite la edición manual de las imágenes. Han sido analizadas 120 imágenes de seis voladuras, elegidas aleatoriamente. Tras el estudio granulométrico, se observa que las curvas granulométricas obtenidas con ambos sistemas, estadísticamente, no son la misma en la mayor parte de la curva, por tanto, se analiza una posible relación entre los tamaños característicos X50 y X80, llegando a la conclusión de que ninguno de los sistemas es totalmente fiable, y es necesario calibrar los sistemas con datos de fragmentación reales obtenidos por medio de básculas. Abstract In mining, the estimate of the granulometric curve blasting debris is very important to evaluate the design, implementation and optimization of it. Currently, for the obtaining of this curves are used digital system of pictures taken by a camera. In this project, the fragmentation of six rock blasting were analyzed. The rock blastings are executed in 2012 in the quarry “El Aljibe” located in Almonacid de Toledo, with a automatic online system (Split Online) and a manual editing software (WipFrag). 120 randomly selected pictures have been analyzed. After the granulometric study, it appears that the size distribution curves obtained with both systems, statistically, are not the same, then, a possible relationship between the feature sizes X50 and X80 is analyzed, concluding that none of the systems is fully reliable, and systems must be calibrated with real data fragmentation obtained from data scales.

El palacio de Diocleciano en Split

Estudio preliminar de la reproducción facsímil del libro El palacio de Diocleciano en Spalato, de Jacques Zeiller, 1912. perteneciente a la serie Monumentos Arquitectónicos de España. Serie : Fondo antiguo de la Escuela Técnica Superior de Arquitectura de Madrid ; 12

Architectural limits on split genes

Exon/intron architecture varies across the eukaryotic kingdom with large introns and small exons the rule in vertebrates and the opposite in lower eukaryotes. To investigate the relationship between exon and intron size in pre-mRNA processing, internally expanded exons were placed in vertebrate genes with small and large introns. Both exon and intron size influenced splicing phenotype. Intron size dictated if large exons were efficiently recognized. When introns were large, large exons were skipped; when introns were small, the same large exons were included. Thus, large exons were incompatible for splicing if and only if they were flanked by large introns. Both intron and exon size became problematic at ≈500 nt, although both exon and intron sequence influenced the size at which exons and introns failed to be recognized. These results indicate that present-day gene architecture reflects at least in part limitations on exon recognition. Furthermore, these results strengthen models that invoke pairing of splice sites during recognition of pre-mRNAs, and suggest that vertebrate consensus sequences support pairing across either introns or exons.

Murine cytomegalovirus M78 protein, a G protein-coupled receptor homologue, is a constituent of the virion and facilitates accumulation of immediate-early viral mRNA

The M78 protein of murine cytomegalovirus exhibits sequence features of a G protein-coupled receptor. It is synthesized with early kinetics, it becomes partially colocalized with Golgi markers, and it is incorporated into viral particles. We have constructed a viral substitution mutant, SMsubM78, which lacks most of the M78 ORF. The mutant produces a reduced yield in cultured 10.1 fibroblast and IC21 macrophage cell lines. The defect is multiplicity dependent and greater in the macrophage cell line. Consistent with its growth defect in cultured cells, the mutant exhibits reduced pathogenicity in mice, generating less infectious progeny than wild-type virus in all organs assayed. SMsubM78 fails to efficiently activate accumulation of the viral m123 immediate-early mRNA in infected macrophages. M78 facilitates the accumulation of the immediate-early mRNA in cycloheximide-treated cells, arguing that it acts in the absence of de novo protein synthesis. We conclude that the M78 G protein-coupled receptor homologue is delivered to cells as a constituent of the virion, and it acts to facilitate the accumulation of immediate-early mRNA.

Use of virion DNA as a cloning vector for the construction of mutant and recombinant herpesviruses.

We have developed improved procedures for the isolation of deletion mutant, point mutant, and recombinant herpesvirus saimiri. These procedures take advantage of the absence of NotI and AscI restriction enzyme sites within the viral genome and use reporter genes for the identification of recombinant viruses. Genes for secreted engineered alkaline phosphatase and green fluorescent protein were placed under simian virus 40 early promoter control and flanked by NotI and AscI restriction sites. When permissive cells were cotransfected with herpesvirus saimiri virion DNA and one of the engineered reporter genes cloned within herpesvirus saimiri sequences, recombinant viruses were readily identified and purified on the basis of expression of the reporter gene. Digestion of recombinant virion DNA with NotI or AscI was used to delete the reporter gene from the recombinant herpesvirus saimiri. Replacement of the reporter gene can be achieved by NotI or AscI digestion of virion DNA and ligation with a terminally matched fragment or, alternatively, by homologous recombination in cotransfected cells. Any gene can, in theory, be cloned directly into the virion DNA when flanked by the appropriate NotI or AscI sites. These procedures should be widely applicable in their general form to most or all herpesviruses that replicate permissively in cultured cells.

Split invertase polypeptides form functional complexes in the yeast periplasm in vivo.

The assembly of functional proteins from fragments in vivo has been recently described for several proteins, including the secreted maltose binding protein in Escherichia coli. Here we demonstrate for the first time that split gene products can function within the eukaryotic secretory system. Saccharomyces cerevisiae strains able to use sucrose produce the enzyme invertase, which is targeted by a signal peptide to the central secretory pathway and the periplasmic space. Using this enzyme as a model we find the following: (i) Polypeptide fragments of invertase, each containing a signal peptide, are independently translocated into the endoplasmic reticulum (ER) are modified by glycosylation, and travel the entire secretory pathway reaching the yeast periplasm. (ii) Simultaneous expression of independently translated and translocated overlapping fragments of invertase leads to the formation of an enzymatically active complex, whereas individually expressed fragments exhibit no activity. (iii) An active invertase complex is assembled in the ER, is targeted to the yeast periplasm, and is biologically functional, as judged by its ability to facilitate growth on sucrose as a single carbon source. These observation are discussed in relation to protein folding and assembly in the ER and to the trafficking of proteins through the secretory pathway.

The virion glycoproteins of Ebola viruses are encoded in two reading frames and are expressed through transcriptional editing.

In late 1994 and early 1995, Ebola (EBO) virus dramatically reemerged in Africa, causing human disease in the Ivory Coast and Zaire. Analysis of the entire glycoprotein genes of these viruses and those of other EBO virus subtypes has shown that the virion glycoprotein (130 kDa) is encoded in two reading frames, which are linked by transcriptional editing. This editing results in the addition of an extra nontemplated adenosine within a run of seven adenosines near the middle of the coding region. The primary gene product is a smaller (50-70 kDa), nonstructural, secreted glycoprotein, which is produced in large amounts and has an unknown function. Phylogenetic analysis indicates that EBO virus subtypes are genetically diverse and that the recent Ivory Coast isolate represents a new (fourth) subtype of EBO virus. In contrast, the EBO virus isolate from the 1995 outbreak in Kikwit, Zaire, is virtually identical to the virus that caused a similar epidemic in Yambuku, Zaire, almost 20 years earlier. This genetic stability may indicate that EBO viruses have coevolved with their natural reservoirs and do not change appreciably in the wild.

Trans-dominant inhibitory human immunodeficiency virus type 1 protease monomers prevent protease activation and virion maturation.

Production of infectious human immunodeficiency virus (HIV) requires proper polyprotein processing by the dimeric viral protease. The trans-dominant inhibitory activity of a defective protease monomer with the active site Asp-25 changed to Asn was measured by transient transfection. A proviral plasmid that included the drug-selectable Escherichia coli gpt gene was used to deliver the wild-type (wt) or mutant proteases to cultured cells. Coexpression of the wt proviral DNA (HIV-gpt) with increasing amounts of the mutant proviral DNA (HIV-gpt D25N) results in a concomitant decrease in proteolytic activity monitored by in vivo viral polyprotein processing. The viral particles resulting from inactivation of the protease were mostly immature, consisting predominantly of unprocessed p55gag and p160gag-pol polyproteins. In the presence of HIV-1 gp160 env, the number of secreted noninfectious particles correlated with the presence of increasing amounts of the defective protease. Greater than 97% reduction in infectivity was observed at a 1:6 ratio of wt to defective protease DNA. This provides an estimate of the level of inhibition required for effectively preventing virion processing. Stable expression of the defective protease in monkey cells reduced the yield of infectious particles from these cells by 90% upon transfection with the wt proviral DNA. These results show that defective subunits of the viral protease exert a trans-dominant inhibitory effect resulting from the formation of catalytically compromised heterodimers in vivo, ultimately yielding noninfectious viral particles.

Mutagenesis of the putative alpha-helical domain of the Vpr protein of human immunodeficiency virus type 1: effect on stability and virion incorporation.

vpr is one of the auxiliary genes of human immunodeficiency virus type 1 (HIV-1) and is conserved in the related HIV-2/simian immunodeficiency virus lentiviruses. The unique feature of Vpr is that it is the only nonstructural protein incorporated into the virus particle. Secondary structural analysis predicted an amphipathic alpha-helical domain in the amino terminus of Vpr (residues 17-34) which contains five acidic and four leucine residues. To evaluate the role of specific residues of the helical domain for virion incorporation, mutagenesis of this domain was carried out. Substitution of proline for any of the individual acidic residues (Asp-17 and Glu-21, -24, -25, and -29) eliminated the virion incorporation of Vpr and also altered the stability of Vpr in cells. Conservative replacement of glutamic residues of the helical domain with aspartic residues resulted in Vpr characteristic of wild type both in stability and virion incorporation, as did substitution of glutamine for the acidic residues. In contrast, replacement of leucine residues of the helical domain (residues 20, 22, 23, and 26) by alanine eliminated virion incorporation function of Vpr. These data indicate that acidic and hydrophobic residues and the helical structure in this region are critical for the stability of Vpr and its efficient incorporation into virus-like particles.