Classification and identification of organic binding media in artworks by means of Fourier transform infrared spectroscopy and principal component analysis.

Fourier transform infrared spectroscopy is a powerful analytical technique to study organic materials. However, in Cultural Heritage, since the sample under analysis is always a complicated matrix of several materials, data analysis performed through peak-by-peak comparisons of sample spectra with those of standard compounds is a tedious method that does not always provide good results. To overcome this problem, a chemometric model based on principal component analysis was developed to classify and identify organic binding media in artworks. The model allows the differentiation of five families of binders: drying oils, waxes, proteins, gums, and resins, taking into account the absorption bands in two characteristic spectral windows: C-H stretching and carbonyl band. This new methodology was applied in the characterization of binders in three kinds of artworks: papers of historical, archeological, and artistic value, easel paintings, and polychromed stone-based sculptures.

Gestión de calidad en el diagnóstico genético preimplantación / Quality management in preimplantation genetic diagnosis

El diagnóstico genético preimplantación (DGP) se haconsolidado como una forma precoz de diagnóstico prenatalrealizada en embriones en estadios previos a la implantación y con el objeto de evitar la transmisión de enfermedades genéticas a la descendencia. La principal característica del DGP es el hecho de adaptar todos los protocolos de diagnóstico genético a una sola célula embrionaria. En este contexto es necesario maximizar la seguridad y la reproducibilidad de los procedimientos. La implantación de un sistema de gestión de la calidad (SGC) en un laboratorio de DGP nos permite evaluar la efectividad de los procedimientos, identificar y corregir errores, asegurar la sensibilidad y la precisiónde los diferentes procesos y supervisar la competencia de los profesionales que trabajan en los diferentes laboratorios. Actualmente estamos asistiendo a la creación de los primeros grupos de expertos implicados en la gestión de la calidad en los laboratorios de DGP y se han publicado las primeras directrices internacionales en este sentido. La implementación de un SGC representa un trabajo adicional y requiere de una fuerte inversión en diferentes áreas. Presentamosla implementación del primer programa de gestiónde la calidad basado en la norma UNE ISO 9001:2000en un servicio de DGP en Europa

Preimplantation genetic diagnosis (PGD) has beenestablished as an early form of prenatal diagnosis. It isperformed in preimplantation stages embryos in order toavoid the transmission of genetic disorders. The principal trait of PGD is the adaptation of diagnosis protocols to only one embryonic cell. In this sense, it is necessary to ensure the safety and reproducibility of all procedures. The implementation of a Quality Management System (QMS) leads the procedure efficiency evaluation, identification and correction of errors, ensure the sensitivity and supervise the competence of the professional teams working at laboratories. Nowadays, we assist to the creation of the first experts groups concerning to quality management in the frame of PGD. First international guidelines have been published, recently. The implementation of a QMS leads a huge amount of additional work and requires a lot of investment in all kind of areas. We present the first PGD quality management program based on UNE ISO 9001:2000 implemented in Europef (AU)

Recomendaciones de calidad para el diagnóstico prenatal molecular / Quality recommenations in molecular prenatal diagnosis

El diagnóstico prenatal molecular consiste en la aplicación de técnicas de genética molecular sobre una muestra fetal obtenida generalmente mediante una técnica invasiva con el objetivo de diagnosticar una determinada enfermedad hereditaria ante un riesgo clínico evidente. Desde que en 1975 se realizara el primer diagnóstico prenatal molecular hasta la fecha, el panorama ha cambiado considerablemente. El descubrimiento y aplicación de nuevas técnicas moleculares, la mejora de las técnicas invasivas con la consiguiente reducción del riesgo de pérdida fetal y, en particular, la gran experiencia adquirida en las unidades multidisciplinaresde genética han convertido al diagnóstico prenatalmolecular en una opción diagnóstica rutinaria con una importante demanda social. Además, el número de enfermedades hereditarias que pueden ser diagnosticadas y el número de mutaciones identificadas en pacientes afectados por enfermedades raras es cada vez mayor, lo que posibilita cada vez más la aplicación de esta opción diagnóstica. Ante esta situación de crecimiento, tanto en cantidad de diagnósticos como en su diversidad y complejidad, y teniendo en cuenta las importantes implicaciones, tanto hereditarias como éticas y sociales, resulta necesario disponer de unos criteriosmínimos de calidad que garanticen un adecuado diagnósticoprenatal y de este modo posibilitar la posterior toma dedecisiones. Estas recomendaciones deben tener en cuentano sólo los aspectos técnicos que rodean el análisis de la muestra de ADN fetal, sino también la planificación previa y la viabilidad de un determinado tipo de diagnóstico, así como la correcta interpretación y comunicación a la pareja de las implicaciones de los resultados obtenidos

The molecular prenatal diagnosis consists in theapplication of molecular genetics techniques on a foetalsample obtained, generally, using an invasive techniqueto diagnose a particular inherited disease when an evident clinical risk is present. Since in 1975 the first molecular prenatal diagnosis was performed up to now, the picture has changed considerably. The discovery and implementation of new molecular techniques, the improving of the invasive techniques with a consequent reduction of the risk for foetal loss and in particular the great experience acquired in the multidisciplinary genetics units have become the molecular prenatal diagnosis in a routine diagnostic option with an important social demand. In addition, the number of inherited diseases that can be diagnosed and the number of mutations identified in patients affected by rare diseases is increasing, enabling more and more the application of this diagnosis option. In this situation of growth, both in quantity of diagnoses as in its diversity and complexity, and taking into account the important implications, not only hereditary but also ethical and social, is necessary to have minimum of quality standards in order to guarantee an adequate prenatal diagnosis and thus enabling the subsequent decision making process. These recommendationsshould take into account, not just the technicalaspects surrounding the analysis of the sample offoetal DNA, but also the planning and the feasibility of a particular type of diagnosis, as well as the correct interpretation and communication of all the results implications to the couple (AU)

Essential oil composition of the different parts of Eryngium corniculatum Lam. (Apiaceae) from Spain.

The essential oil from the different parts (inflorescences, stems+leaves and roots) of E. corniculatum Lam. gathered in Guadalajara (Spain) has been extracted by steam distillation and analysed by gas chromatography and gas chromatography coupled to mass spectrometry. Quantitative and qualitative differences have been found between the analysed parts, although all of them contained the same principal compound, 2,4,6-trimethylbenzaldehyde, representing the 50.8%, 50.0%, and 29.8% of the total oil for inflorescences, stems+leaves and roots, respectively. Other representative constituents of the oil were similar in the different fractions: in the inflorescences compounds were found to be alpha-pinene (4.0%), chrysanthenyl acetate (4.0%), 2,4,5-trimethylbenzaldehyde (3.3%), (2Z,6E)-farnesol (2.0%), (E)-nerolidol (2.1%) and (Z)-beta-ocimene (2.1%), while the stems+leaves oil showed 2,4,5-trimethylbenzaldehyde (3.8%), alpha-pinene (3.4%), (E)-nerolidol (2.4%) and (2Z,6E)-farnesol (2.1%), and in the roots oil a phyllocladene isomer (13.0%), (E)-nerolidol (9.4%), beta-eudesmol (4.1%) and (2Z,6E)-farnesol (2.1%) were found. The presence of C-10 compounds as the main fraction for an Eryngium species is worth mentioning. This is the first report on the chemical composition of this Mediterranean endemic species.

Analysis by gas chromatography-mass spectrometry of the volatiles from the fruits of Ammodaucus leucotrichus subsp. leucotrichus and subsp. nanocarpus grown in North Africa and the Canary Islands, respectively.

The volatiles from the fruits of Ammodaucus leucotrichus subsp. leucotrichus and subsp. nanocarpus (two endemic species, the first from North Africa and the second from the Canary Islands, Spain) were studied by gas chromatography and gas chromatography-mass spectrometry. The major components of the volatiles of subsp. nanocarpus were found to be, beta-pinene (22.2-33.6%), bornyl angelate (20.6-21.8%) and camphor (8.3-11.7%) whereas in the fruits of subsp. leucotrichus, the main constituents were perillaldehyde (63.6%) and limonene (26.8%). We also suggest that subsp. nanocarpus should have the status of species and should be named Ammodaucus nanocarpus.

Analysis by gas chromatography-mass spectrometry of the essential oils from the aerial parts of Pimpinella anagodendron Bolle and Pimpinella rupicola Svent., two endemic species to the Canary Islands, Spain.

The essential oils from the aerial parts of Pimpinella anagodendron Bolle and Pimpinella rupicola Svent., two endemic species growing in Tenerife, Canary Islands, Spain, were studied by gas chromatography and gas chromatography-mass spectrometry. The major components of the flowering tops (flowers+unripe fruits) of P. rupicola (PRFT) were found to be beta-bisabolene (34.8%), limonene (10.9%) and alpha-zingiberene (10.5%), whereas in the flowering tops of P. anagodendron (PAFT), the main constituents were alpha-zingiberene (32.9%), beta-bisabolene (17.9%), beta-pinene (15.8%) and ar-curcumene (11.5%). The major compounds found in the stems+leaves of P. rupicola (PRSL) were beta-bisabolene (31.6%), alpha-zingiberene (11.4%) and limonene (10.8%), whereas those of P. anagodendron (PASL) were alpha-zingiberene (32.3%), beta-bisabolene (14.0%) and ar-curcumene (12.6%). In all the oils were found the characteristic constituents of genus Pimpinella, the pseudoisoeugenol esters. In accordance with the morphological, chorological and chemical differences between both species, we suggest that P. rupicola Svent. is a good taxon and not a synonym of P. anagodendron.

On-line FT-Raman and dispersive Raman spectra database of artists' materials (e-VISART database).

Raman spectroscopy has been widely applied in the analysis of different types of artwork. This technique is sensitive, reliable, non-destructive and can be used in situ. However, there are few references in the literature regarding specific Raman spectra libraries for the field of artwork analysis. In this paper, the development of two on-line databases with Fourier transform Raman (FT-Raman; 1064 nm) and dispersive Raman (785 nm) spectra of materials used in fine art is presented; both are implemented in the e-vibrational spectroscopic databases of artists' materials database (e-VISART). The database provides not only spectra, but also information about each pigment. It must be highlighted that for each pigment or material several spectra are available from different dealers. Some of the FT-Raman spectra available in the e-VISART database have not been published until now. Some examples in which the e-VISART database has been successfully used are presented.

Raman fibre optic approach to artwork dating.

Raman micro-probe spectroscopy has been applied to the analysis of a non catalogued hand-crafted wallpaper during its restoration process. The analysis has been totally non-destructive without the necessity of taking any sample. The artwork showed a great chromatic palette having been detected the presence of calcium carbonate, Prussian blue, ultramarine blue, gypsum (CaSO4.2H2O), minium (Pb3O4), vermilion (HgS), chrome orange (CaCO3), chrome yellow (PbCrO4), barium sulphate and carbon black (C). From the spectroscopic analysis the date of its manufacturing has been set between 1828 and 1830, introduction of chrome yellow and orange, as well as artificial ultramarine blue, and 1840, when continuous industrial wallpapers were extensively manufactured in Europe.

Essential oil composition of the leaves and stems of Meum athamanticum Jacq., from Spain.

The essential oil of the leaves and stems of Meum athamanticum Jacq., has been extracted by steam distillation and analysed by gas chromatography (GC) and gas chromatography coupled to mass spectrometry (GC-MS). The monoterpene fraction was predominant while the sesquiterpene one was practically absent. The principal constituents have been identified as (E)-beta-ocimene (29.6%), gamma-terpinene (17.9%), terpinolene (17.0%) and p-cymene (9.7%). Our results show that the chemical composition of the essential oil obtained of the leaves and stems of M. athamanticum from Spain is different to that obtained from plants of Germany, Italy and France.

Analysis of bulk and inorganic degradation products of stones, mortars and wall paintings by portable Raman microprobe spectroscopy.

This work reports the use of a portable Raman microprobe spectrometer for the analysis of bulk and decaying compounds in carbonaceous materials such as stones, mortars and wall paintings. The analysed stones include limestone, dolomite and carbonaceous sandstone, gypsum and calcium oxalate, both mono- and dihydrated, being the main inorganic degradation products detected. Mortars include bulk phases with pure gypsum, calcite and mixtures of both or with sand, soluble salts being the most important degradation products. The pigments detected in several wall paintings include Prussian blue, iron oxide red, iron oxide yellow, vermilion, carbon black and lead white. Three different decaying processes have been characterised in the mortars of the wall paintings: (a) a massive absorption of nitrates that reacted with calcium carbonate and promoted the unbinding of pigment grains, (b) the formation of black crusts in the vault of the presbytery and (c) the thermodecomposition of pigments due to a fire.

Analysis by gas chromatography-mass spectrometry of the essential oil from the aerial parts of Pimpinella junoniae Ceb. & Ort., gathered in La Gomera, Canary Islands, Spain.

The essential oil from the aerial parts of Pimpinella junoniae Ceb. & Ort., growing in La Gomera, Canary Islands, Spain, was studied by gas chromatography and gas chromatography-mass spectrometry, and 43 constituents were identified. The major components were found to be alpha-zingiberene (20.6%), alpha-pinene (17.9%), (E)-beta-farnesene (9.3%), ar-curcumene (7.4%), beta-phellandrene (7.0%), beta-bisabolene (6.1%) and epoxypseudoisoeugenyl 2-methylbutyrate (6.0%). The decomposition product of epoxypseudoisoeugenol derivatives, 5-methoxy-2-methylbenzofuran (5.7%), moderate amounts of other arylpropanoids with the pseudoisoeugenol skeleton (total percentage, 5.2%) and other compounds such as beta-sesquiphellandrene (3.0%), cis-beta-guaiene (1.5%), alpha-phellandrene (1.5%) and alpha-bisabolol (1.3%), were also found.

Analysis by gas chromatography-mass spectrometry of the essential oils from the aerial parts of Rutheopsis herbanica (Bolle) Hans. & Kunk., gathered in Fuerteventura (Canary Islands).

The essential oil from the aerial parts of Rutheopsis herbanica (Bolle) Hans. & Kunk., growing in Fuerteventura, Canary Islands, Spain, was studied by gas chromatography and gas chromatography-mass spectrometry, and 42 constituents were identified. The major components were found to be alpha-pinene (29.4%), dillapiole (21.3%), limonene (14.1%), beta-pinene (13.2%) and myristicin (10.0%). As far as we know, this is the first report on the essential oil composition of this species.

Analysis by gas chromatography-mass spectrometry of the volatile components of Ageratina adenophora Spreng., growing in the Canary Islands.

The essential oil of Ageratina adenophora Spreng., growing in the Canary Islands was analyzed by GC-MS. A total of 78 volatile compounds was identified and p-cymene (11.6%) was the major component in the oil. The sesquiterpene fraction (44.3%) was higher than the monoterpene one (32.1%).

Analysis of the volatile components of Argyranthemum adauctum (Link.) Humphries by gas chromatography-mass spectrometry.

The volatile constituents of the essential oil of Argyranthemum adauctum (Link.) Humphries extracted by steam distillation have been studied by GC-MS. A total of 60 compounds have been identified representing the 89.8% of the oil. The major constituents were found to be beta-pinene (27.4%) and santolinatriene (22.6%). This is the first report on the chemical compounds of the oil of this species.

Seasonal variation in chemical constituents of Santolina rosmarinifolia L. ssp. rosmarinifolia.

The seasonal variation of the essential oil extracts from the aerial parts of a Santolina rosmarinifolia population has been studied. Oil yields increased in the months of March, April, May and June. Oil concentration showed significant correlations with both precipitation (positive) and temperature (negative). Essential oil main components were sabinene, beta-pinene, myrcene, beta-phellandrene, limonene, 1,8-cineole, artemisia ketone, terpinen-4-ol, capillene, ar-curcumene and beta-eudesmol. Monoterpenes like beta-phellandrene, limonene and 1,8-cineole showed a significant negative correlation with temperature, while capillene presented a strong positive correlation with precipitation. The rest of the essential oil components did not show any noticeable trend.

Structural and evolutionary analysis of the copia-like elements in the Arabidopsis thaliana genome.

The analysis of 460 kb of genomic sequence of Arabidopsis thaliana chromosome III allowed us to identify two new transposable elements named AtC1 and AtC2. AtC1 shows identical long terminal repeats (LTRs) and all the structural features characteristic of the copia-like active elements. AtC2 is also a full copia-like element, but a putative stop codon in the open reading frame (ORF) would produce a truncated protein. In order to identify the copia-like fraction of the A. thaliana genome, a careful computer-based analysis of the available sequences (which correspond to 92% of the genome) was performed. Approximately 300 nonredundant copia-like sequences homologous to AtC1 and AtC2 were detected, which showed an extreme heterogeneity in size and degree of conservation. This number of copies would correspond to approximately 1% of the A. thaliana genome. Seventy-one sequences were selected for further analysis, with 23 of them being full complete elements. Five corresponded to previously described ones, and the remaining ones, named AtC3 to AtC18 are new elements described in this work. Most of these elements presented a putative functional ORF, nearly identical LTRs, and the other elements necessary for retrotransposon activity. Phylogenetic trees, supported by high bootstrap values, indicated that these 23 elements could be considered separate families. In turn, these 23 families could be clustered into six major lineages, named copia I-VI. Most of the 71 analyzed sequences clustered into these six main clades. The widespread presence of these copia-like superfamilies throughout plant genomes is discussed.

Sequence and analysis of chromosome 3 of the plant Arabidopsis thaliana.

Arabidopsis thaliana is an important model system for plant biologists. In 1996 an international collaboration (the Arabidopsis Genome Initiative) was formed to sequence the whole genome of Arabidopsis and in 1999 the sequence of the first two chromosomes was reported. The sequence of the last three chromosomes and an analysis of the whole genome are reported in this issue. Here we present the sequence of chromosome 3, organized into four sequence segments (contigs). The two largest (13.5 and 9.2 Mb) correspond to the top (long) and the bottom (short) arms of chromosome 3, and the two small contigs are located in the genetically defined centromere. This chromosome encodes 5,220 of the roughly 25,500 predicted protein-coding genes in the genome. About 20% of the predicted proteins have significant homology to proteins in eukaryotic genomes for which the complete sequence is available, pointing to important conserved cellular functions among eukaryotes.

Progress in Arabidopsis genome sequencing and functional genomics.

Arabidopsis thaliana has a relatively small genome of approximately 130 Mb containing about 10% repetitive DNA. Genome sequencing studies reveal a gene-rich genome, predicted to contain approximately 25000 genes spaced on average every 4.5 kb. Between 10 to 20% of the predicted genes occur as clusters of related genes, indicating that local sequence duplication and subsequent divergence generates a significant proportion of gene families. In addition to gene families, repetitive sequences comprise individual and small clusters of two to three retroelements and other classes of smaller repeats. The clustering of highly repetitive elements is a striking feature of the A. thaliana genome emerging from sequence and other analyses.

ZFWD: a novel subfamily of plant proteins containing a C3H zinc finger and seven WD40 repeats.

We describe a new subfamily of WD repeat proteins characterised by the presence of a C3H zinc finger at the N-terminal part of the protein associated with seven WD40 repeats. We have identified four members of this subfamily in Arabidopsis thaliana, one of them with associated expressed sequence tags (ESTs). We have also identified homologous ESTs in rice, cotton, maize, poplar, pine tree and the ice plant. We do not observe animal homologues, suggesting that this subfamily could be specific for plants. Our data suggest an important role for these proteins. Based on the high sequence conservation within the conserved domains, we suggest that these proteins could have a regulatory function.