Cardiovascular imaging 2011 in the International Journal of Cardiovascular Imaging

Cardiovascular Aspects of Radiolog

Aproximación a dos problemas práctico referidos a la Ley de Marcas

<p>En breves líneas nos proponemos abordar dos problemas de naturaleza evidentemente práctica, que creemos pasan desapercibidos en la persecución penal de los delitos previstos en la Ley N° 17.0111 , cuando la titular del registro involucrado es una persona jurídica domiciliada en el extranjero.</p><p>Contenido: Introducción. La instancia de parte en los delitos marcarios. Consecuencias del incumplimento. Los presupuestos para la aplicación del artículo 83 a mercaderías extranjeras. Conclusiones</p

<i>SNaPAfu</i>: A Novel Single Nucleotide Polymorphism Multiplex Assay for <i>Aspergillus fumigatus</i> Direct Detection, Identification and Genotyping in Clinical Specimens

<div><p>Objective</p><p>Early diagnosis of invasive aspergillosis is essential for positive patient outcome. Likewise genotyping of fungal isolates is desirable for outbreak control in clinical setting. We designed a molecular assay that combines detection, identification, and genotyping of <i>Aspergillus fumigatus</i> in a single reaction.</p><p>Methods</p><p>To this aim we combined 20 markers in a multiplex reaction and the results were seen following mini-sequencing readings. Pure culture extracts were firstly tested. Thereafter, <i>Aspergillus</i>-DNA samples obtained from clinical specimens of patients with possible, probable, or proven aspergillosis according to European Organization for the Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG) criteria.</p><p>Results</p><p>A new set of designed primers allowed multilocus sequence typing (MLST) gene amplification in a single multiplex reaction. The newly proposed <i>SNaPAfu</i> assay had a specificity of 100%, a sensitivity of 89% and detection limit of 1 ITS copy/mL (∼0.5 fg genomic <i>Aspergillus</i>-DNA/mL). The marker A49_F was detected in 89% of clinical samples. The <i>SNaPAfu</i> assay was accurately performed on clinical specimens using only 1% of DNA extract (total volume 50 µL) from 1 mL of used bronchoalveolar lavage.</p><p>Conclusions</p><p>The first highly sensitive and specific, time- and cost-economic multiplex assay was implemented that allows detection, identification, and genotyping of <i>A. fumigatus</i> strains in a single amplification followed by mini-sequencing reaction. The new test is suitable to clinical routine and will improve patient management.</p></div

Clinical samples considered in this study and methods used for diagnosis of invasive aspergillosis (ARDS: Acute Respiratory Destress Syndrome; BAL: bronchoalveolar lavages; BS: skin biopsy samples; nm: not enough material to be tested; GM: galactomannan; PF-PCR: Pan-fungal PCF; (+) and (−) represent positive and negative results).

<p>Clinical samples considered in this study and methods used for diagnosis of invasive aspergillosis (ARDS: Acute Respiratory Destress Syndrome; BAL: bronchoalveolar lavages; BS: skin biopsy samples; nm: not enough material to be tested; GM: galactomannan; PF-PCR: Pan-fungal PCF; (+) and (−) represent positive and negative results).</p

The primers used in <i>SNaPAfu</i> assay.

#<p>Primer nomenclature and incorporated information on the targeting polymorphic position (e.g. A45_R means that a reverse primer was designed in the polymorphic position 45 of the gene ANXC4 of MLST panel).</p>§<p>Expected peak size in the electropherogram (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075968#pone-0075968-g002" target="_blank">Figure 2</a>); bp means base pairs.</p>*<p>Base expected in MLST and <i>SNaP</i> profiles; a complementary base is expected to be seen on the <i>SNaPAfu</i> electropherogram for primers designed for reverse sequence.</p

Primers used for MLST genotyping of <i>Aspergillus fumigatus</i> in a multiplex reaction.

*<p>Primer used for sequencing.</p

<i>SNaPAfu</i> assay: A) position of each marker on the automated electropherogram; and B) example of an <i>Aspergillus fumigatus</i> profile (peaks: orange – ladder; blue – guanine; black – cytosine; green – adenine; red – thymine; the interpretation of such the peaks gives the electropherogram reading, then converted to the intermediate profile where the results of the markers with the reverse primer were converted in the complementary base – markers marked bold were converted); and C) the <i>SNaP</i> profile of the isolate presented above obtained according to <b>Table 2</b> (e.g. A45_R, A49_F, … Z198_F) to facilitate comparison with MLST data – markers are presented in alphabetic order; when markers are amplified using reverse primers (e.g. A45_R) the complementary base should be included in the <i>SNaP</i> profile.

<p><i>SNaPAfu</i> assay: A) position of each marker on the automated electropherogram; and B) example of an <i>Aspergillus fumigatus</i> profile (peaks: orange – ladder; blue – guanine; black – cytosine; green – adenine; red – thymine; the interpretation of such the peaks gives the electropherogram reading, then converted to the intermediate profile where the results of the markers with the reverse primer were converted in the complementary base – markers marked bold were converted); and C) the <i>SNaP</i> profile of the isolate presented above obtained according to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075968#pone-0075968-t002" target="_blank"><b>Table 2</b></a> (e.g. A45_R, A49_F, … Z198_F) to facilitate comparison with MLST data – markers are presented in alphabetic order; when markers are amplified using reverse primers (e.g. A45_R) the complementary base should be included in the <i>SNaP</i> profile.</p

Graphical Modeling of Gene Expression in Monocytes Suggests Molecular Mechanisms Explaining Increased Atherosclerosis in Smokers

<div><p>Smoking is a risk factor for atherosclerosis with reported widespread effects on gene expression in circulating blood cells. We hypothesized that a molecular signature mediating the relation between smoking and atherosclerosis may be found in the transcriptome of circulating monocytes. Genome-wide expression profiles and counts of atherosclerotic plaques in carotid arteries were collected in 248 smokers and 688 non-smokers from the general population. Patterns of co-expressed genes were identified by Independent Component Analysis (ICA) and network structure of the pattern-specific gene modules was inferred by the PC-algorithm. A likelihood-based causality test was implemented to select patterns that fit models containing a path “smoking→gene expression→plaques”. Robustness of the causal inference was assessed by bootstrapping. At a FDR ≤0.10, 3,368 genes were associated to smoking or plaques, of which 93% were associated to smoking only. <em>SASH1</em> showed the strongest association to smoking and <em>PPARG</em> the strongest association to plaques. Twenty-nine gene patterns were identified by ICA. Modules containing <em>SASH1</em> and <em>PPARG</em> did not show evidence for the “smoking→gene expression→plaques” causality model. Conversely, three modules had good support for causal effects and exhibited a network topology consistent with gene expression mediating the relation between smoking and plaques. The network with the strongest support for causal effects was connected to plaques through <em>SLC39A8</em>, a gene with known association to HDL-cholesterol and cellular uptake of cadmium from tobacco, while smoking was directly connected to <em>GAS6</em>, a gene reported to have anti-inflammatory effects in atherosclerosis and to be up-regulated in the placenta of women smoking during pregnancy. Our analysis of the transcriptome of monocytes recovered genes relevant for association to smoking and atherosclerosis, and connected genes that before, were only studied in separate contexts. Inspection of correlation structure revealed candidates that would be missed by expression-phenotype association analysis alone.</p> </div