Summary
Background Most recently, a new rapid and fully automated electrochemiluminescence immunoassay for the determination of TSH receptor autoantibodies (TRAb) based on the ability of TRAb to ...inhibit the binding of a human thyroid‐stimulating monoclonal antibody (M22) has been established.
Objective To evaluate this assay system in clinical routine based on an international multicentre trial and to compare the results with other established TRAb assays.
Patients and measurements Totally 508 Graves’ disease (GD), 142 autoimmune thyroiditis, 107 subacute thyroiditis, 109 nonautoimmune nodular goitre, 23 thyroid cancer patients and 446 normal controls were retrospectively evaluated.
Results ROC plot analysis revealed an area under curve of 0·99 (95% CI: 0·99–1·0) indicating a high assay sensitivity and specificity. The highest sensitivity (99%) and specificity (99%) was seen at a cut‐off level of 1·75 IU/l. Here, the calculated positive predictive value was 95%, whereas the negative predictive value was 100%. Applying the ROC plot‐derived cut‐off of 1·75 IU/l we found a sensitivity for TRAb positivity within the group of newly diagnosed GD patients of 97% which is in accordance to the sum of different nonautomated porcine TSH receptor‐based assays with a sensitivity of 94% indicating an excellent analytical performance of the new assay format. Detailed comparison of the automated and the sum of manual assays revealed a near identical specificity.
Conclusion Our results demonstrate that this new assay system has a high sensitivity for detecting GD and specificity for discriminating from other thyroid diseases. This assay may represent the future technology for rapid fully automated TRAb detection.
El cineasta chino Wong Kar-wai ha reconocido la influencia de las novelas de Manuel Puig en la estructura de sus películas, y esta influencia ha sido comentada por diversos críticos. Sin embargo, se ...ha tendido a obviar el hecho de que el director también presentó Happy Together como “basada en” la novela The Buenos Aires Affair del mismo escritor. Nuestro artículo analiza la influencia de la narrativa de Manuel Puig en las producciones de Wong Kar-wai, haciendo especial hincapié en la que afecta al contenido de los filmes y explica, tal como argumentamos, la construcción de una determinada idea de lo latino, estereotipada pero al mismo tiempo bien definida, que el director contrapone a la de lo chino.
El estudio de la sexualidad desde una mirada social y cultural ha adquirido una gran relevancia en las ciencias sociales y humanas durante las últimas décadas. Este hecho se ha visto reflejado en un ...creciente número de etnografías y análisis antropológicos que dotan de nuevos significados a la diversidad sexual y de género más allá de las lecturas clínicas y biomédicas. En este artículo se ha efectuado una revisión bibliográfica exhaustiva con el objetivo de analizar el recorrido, las transformaciones y los ámbitos de interés de los trabajos sobre diversidad sexo-genérica en la antropología española desde 1980 hasta 2017. Veremos que, actualmente, esta especialidad disciplinaria cuenta con una amplia variedad de temáticas, ya que aparte de los estudios sobre la homosexualidad han florecido nuevas áreas de interés, tales como los estudios trans e intersex. Con este trabajo se pretende contribuir a la construcción de la historia de la antropología de la diversidad sexual y de género española desde una perspectiva situada. Este artículo demuestra que, a pesar de la inequívoca influencia internacional en el desarrollo de la especialidad en España, también existe un recorrido propio fuertemente influenciado por el contexto socio-histórico y la antropología feminista del país.
An annual seasonal cycle of composition of a bacterioplankton community in an oligotrophic coastal system was studied by denaturing gradient gel electrophoresis (DGGE) using five different primer ...sets. Analysis of DGGE fingerprints showed that primer set 357fGC-907rM grouped samples according to seasons. Additionally, we used the set of 16S rRNA genes archived in the RDPII database to check the percentage of perfect matches of each primer for the most abundant bacterial groups inhabiting coastal plankton communities. Overall, primer set 357fGC-907rM was the most suitable for the routine use of PCR-DGGE analyses in this environment.
Summary
The bacterial community composition of activated sludge from a wastewater treatment plant (Almería, Spain) with the particularity of using seawater was investigated by applying ...454‐pyrosequencing. The results showed that Deinococcus‐Thermus, Proteobacteria, Chloroflexi and Bacteroidetes were the most abundant retrieved sequences, while other groups, such as Actinobacteria, Chlorobi, Deferribacteres, Firmicutes, Planctomycetes, Spirochaetes and Verrumicrobia were reported at lower proportions. Rarefaction analysis showed that very likely the diversity is higher than what could be described despite most of the unknown microorganisms probably correspond to rare diversity. Furthermore, the majority of taxa could not be classified at the genus level and likely represent novel members of these groups. Additionally, the nitrifiers in the sludge were characterized by pyrosequencing the amoA gene. In contrast, the nitrifying bacterial community, dominated by the genera Nitrosomonas, showed a low diversity and rarefaction curves exhibited saturation. These results suggest that only a few populations of low abundant but specialized bacteria are responsible for removal of ammonia in these saline wastewater systems.
The bacterial community composition of activated sludge from a seawater‐processing WWTP (Almería, Spain) was investigated by 454‐pyrosequencing. Members of Deinococcus‐Thermus, Proteobacteria, Chloroflexi and Bacteroidetes were the most abundant, and rarefaction curves showed that diversity was higher than what could be described despite most of the unknown microorganisms probably correspond to rare diversity. Additionally, the nitrifying bacterial community was dominated by the genera Nitrosomonas.
In plants, 3‐deoxy‐d‐manno‐oct‐2‐ulosonic acid (Kdo) is a monosaccharide that is only found in the cell wall pectin, rhamnogalacturonan‐II (RG‐II). Incubation of 4‐day‐old light‐grown Arabidopsis ...seedlings or tobacco BY‐2 cells with 8‐azido 8‐deoxy Kdo (Kdo‐N₃) followed by coupling to an alkyne‐containing fluorescent probe resulted in the specific in muro labelling of RG‐II through a copper‐catalysed azide–alkyne cycloaddition reaction. CMP‐Kdo synthetase inhibition and competition assays showing that Kdo and D‐Ara, a precursor of Kdo, but not L‐Ara, inhibit incorporation of Kdo‐N₃ demonstrated that incorporation of Kdo‐N₃ occurs in RG‐II through the endogenous biosynthetic machinery of the cell. Co‐localisation of Kdo‐N₃ labelling with the cellulose‐binding dye calcofluor white demonstrated that RG‐II exists throughout the primary cell wall. Additionally, after incubating plants with Kdo‐N₃ and an alkynated derivative of L‐fucose that incorporates into rhamnogalacturonan I, co‐localised fluorescence was observed in the cell wall in the elongation zone of the root. Finally, pulse labelling experiments demonstrated that metabolic click‐mediated labelling with Kdo‐N₃ provides an efficient method to study the synthesis and redistribution of RG‐II during root growth.
There are few genomic tools available in melon (Cucumis melo L.), a member of the Cucurbitaceae, despite its importance as a crop. Among these tools, genetic maps have been constructed mainly using ...marker types such as simple sequence repeats (SSR), restriction fragment length polymorphisms (RFLP) and amplified fragment length polymorphisms (AFLP) in different mapping populations. There is a growing need for saturating the genetic map with single nucleotide polymorphisms (SNP), more amenable for high throughput analysis, especially if these markers are located in gene coding regions, to provide functional markers. Expressed sequence tags (ESTs) from melon are available in public databases, and resequencing ESTs or validating SNPs detected in silico are excellent ways to discover SNPs.
EST-based SNPs were discovered after resequencing ESTs between the parental lines of the PI 161375 (SC) x 'Piel de sapo' (PS) genetic map or using in silico SNP information from EST databases. In total 200 EST-based SNPs were mapped in the melon genetic map using a bin-mapping strategy, increasing the map density to 2.35 cM/marker. A subset of 45 SNPs was used to study variation in a panel of 48 melon accessions covering a wide range of the genetic diversity of the species. SNP analysis correctly reflected the genetic relationships compared with other marker systems, being able to distinguish all the accessions and cultivars.
This is the first example of a genetic map in a cucurbit species that includes a major set of SNP markers discovered using ESTs. The PI 161375 x 'Piel de sapo' melon genetic map has around 700 markers, of which more than 500 are gene-based markers (SNP, RFLP and SSR). This genetic map will be a central tool for the construction of the melon physical map, the step prior to sequencing the complete genome. Using the set of SNP markers, it was possible to define the genetic relationships within a collection of forty-eight melon accessions as efficiently as with SSR markers, and these markers may also be useful for cultivar identification in Occidental melon varieties.
Melon (Cucumis melo), an economically important vegetable crop, belongs to the Cucurbitaceae family which includes several other important crops such as watermelon, cucumber, and pumpkin. It has ...served as a model system for sex determination and vascular biology studies. However, genomic resources currently available for melon are limited.
We constructed eleven full-length enriched and four standard cDNA libraries from fruits, flowers, leaves, roots, cotyledons, and calluses of four different melon genotypes, and generated 71,577 and 22,179 ESTs from full-length enriched and standard cDNA libraries, respectively. These ESTs, together with ~35,000 ESTs available in public domains, were assembled into 24,444 unigenes, which were extensively annotated by comparing their sequences to different protein and functional domain databases, assigning them Gene Ontology (GO) terms, and mapping them onto metabolic pathways. Comparative analysis of melon unigenes and other plant genomes revealed that 75% to 85% of melon unigenes had homologs in other dicot plants, while approximately 70% had homologs in monocot plants. The analysis also identified 6,972 gene families that were conserved across dicot and monocot plants, and 181, 1,192, and 220 gene families specific to fleshy fruit-bearing plants, the Cucurbitaceae family, and melon, respectively. Digital expression analysis identified a total of 175 tissue-specific genes, which provides a valuable gene sequence resource for future genomics and functional studies. Furthermore, we identified 4,068 simple sequence repeats (SSRs) and 3,073 single nucleotide polymorphisms (SNPs) in the melon EST collection. Finally, we obtained a total of 1,382 melon full-length transcripts through the analysis of full-length enriched cDNA clones that were sequenced from both ends. Analysis of these full-length transcripts indicated that sizes of melon 5' and 3' UTRs were similar to those of tomato, but longer than many other dicot plants. Codon usages of melon full-length transcripts were largely similar to those of Arabidopsis coding sequences.
The collection of melon ESTs generated from full-length enriched and standard cDNA libraries is expected to play significant roles in annotating the melon genome. The ESTs and associated analysis results will be useful resources for gene discovery, functional analysis, marker-assisted breeding of melon and closely related species, comparative genomic studies and for gaining insights into gene expression patterns.
Melon (Cucumis melo) is a horticultural specie of significant nutritional value, which belongs to the Cucurbitaceae family, whose economic importance is second only to the Solanaceae. Its small ...genome of approx. 450 Mb coupled to the high genetic diversity has prompted the development of genetic tools in the last decade. However, the unprecedented existence of a transcriptomic approaches in melon, highlight the importance of designing new tools for high-throughput analysis of gene expression.
We report the construction of an oligo-based microarray using a total of 17,510 unigenes derived from 33,418 high-quality melon ESTs. This chip is particularly enriched with genes that are expressed in fruit and during interaction with pathogens. Hybridizations for three independent experiments allowed the characterization of global gene expression profiles during fruit ripening, as well as in response to viral and fungal infections in plant cotyledons and roots, respectively. Microarray construction, statistical analyses and validation together with functional-enrichment analysis are presented in this study.
The platform validation and enrichment analyses shown in our study indicate that this oligo-based microarray is amenable for future genetic and functional genomic studies of a wide range of experimental conditions in melon.
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