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Background and objective: We aimed to evaluate the safety and outcomes of thrombectomy in anterior circulation acute ischaemic stroke recorded in the SITS–International Stroke Thrombectomy Register (SITS-ISTR) and compare them with pooled randomized controlled trials (RCTs) and two national registry studies. Methods: We identified centres recording ≥10 consecutive patients in the SITS-ISTR with at least 70% of available modified Rankin Scale (mRS) at 3 months during 2014–2019. We defined large artery occlusion as intracranial internal carotid artery, first and second segment of middle cerebral artery and first segment of anterior cerebral artery. Outcome measures were functional independence (mRS score 0-2) and death at 3 months and symptomatic intracranial haemorrhage (SICH) per modified SITS-MOST. Results: Results are presented in the following order: SITS-ISTR, RCTs, MR CLEAN Registry and German Stroke Registry (GSR). Median age was 73, 68, 71 and 75 years; baseline NIHSS score was 16, 17, 16 and 15; prior intravenous thrombolysis was 62%, 83%, 78% and 56%; onset to reperfusion time was 289, 285, 267 and 249 min; successful recanalization (mTICI score 2b or 3) was 86%, 71%, 59% and 83%; functional independence at 3 months was 45.5% (95% CI: 44–47), 46.0% (42–50), 38% (35–41) and 37% (35–41), respectively; death was 19.2% (19–21), 15.3% (12.7–18.4), 29.2% (27–32) and 28.6% (27–31); and SICH was 3.6% (3–4), 4.4% (3.0–6.4), 5.8% (4.7–7.1) and not available. Conclusion: Thrombectomy in routine clinical use registered in the SITS-ISTR showed safety and outcomes comparable to RCTs, and better functional outcomes and lower mortality than previous national registry studies. © 2021 The Authors. Journal of Internal Medicine published by John Wiley & Sons Ltd on behalf of Association for Publication of The Journal of Internal Medicine

Virtual laboratories (VL) and interactive simulations are excellent approaches for training students to understand technical principles. This can be useful in many fields of science and engineering teaching. For this purpose, we have developed a virtual environment that can simulate real-laboratory operations, effectively enhancing the teaching process with an intuitive and appealing interface. Such software shows the virtual-studio practice characterization of basic biofuel-properties, simulating reality step-by-step. This virtual laboratory has been used by students of the postgraduate master subject on Biomass for Power Generation, belonging to the Master of Distributed Renewable Energy. The aim of this tool is to help students to study, learn and investigate on their own. The virtual lab is made of a web-based application to complement experimental laboratory training, allowing students to prepare their experimental practices before going to the lab, and to review them at any time afterwards. The computer application exhibits key virtual-lab educational features, like an integrative layout and self-evaluating tests. It allows a personalized and active learning-process, adaptability to teacher's aims and versatility and simplicity, using different multimedia resources. A satisfaction questionnaire was carried out between master degree students to evaluate the usefulness of the VL. Such a tool was positively evaluated, achieving a mean score of 6 out of 7 points. Additionally, the VL efficiency in the learning process showed a final examination main score of 8 out of 10 points. Last but not least, most students considered that the VL promoted learning and personal effort, being an excellent preparatory tool to real experiments.

Humans have identified and classified biological entities (virusoids, viroids, prokaryotes and eukaryotes) using morphological characteristics. The scientific and technological developments have also allowed to use molecules (molecular markers). The identification and classification of biological entities is important, since it allows its study, control and exploitation. Initially, the peptides were used for such characterization (mainly as isoenzymatic polymorphisms). Currently, the DNA is the molecule of choice. It was first analyzed by RFLP, but the development of the PCR allowed to use such powerful methodology to design faster fingerprinting approaches like RAPD, microsatellites (SSR or STR), AFLP and SNP). The RNA can also be used as a molecular marker. Other powerful technologies for fingerprinting are arrays and microarrays of DNA, RNA or peptides. It is also possible to sequence the genome and transcriptome, which brings these tools to unprecedented levels. Indeed, the DNA, RNA and peptide sequencing are becoming the ultimate molecular markers, with special emphasis on the DNA. As with the genome and transcriptome sequencing, there are many relevant applications of the molecular markers. In fact, the genome may be exploited as a magnificent tool to generate molecular markers, which can be then tested, evaluated and applied in biotechnological developments, including the biomedical sciences.

The common fig tree (Ficus carica L.) is a Mediterranean crop with problematic cultivar identification. The recovery and conservation of possible local varieties for ecological production requires the previous genetic characterization of the available germplasm. In this context, 42 lines corresponding to 12 local varieties and two caprifigs, in addition to 15 reference samples have been fingerprinted using 21 SSR markers. A total of 77 alleles were revealed, detecting a useful level of genetic variability within the local germplasm pools. UPGMA clustering analysis has revealed the genetic structure and relationships among the local and reference germplasm. Eleven of the local varieties could be identified and defined as obtained clusters, showing that SSR analysis is an efficient method to evaluate the Andalusian fig tree diversity for on-farm conservation.

Though a considerable number of publications about biodiesel can be found in literature, several problems remain unsolved, encompassing economical, social, and technical issues. Thus, the biodiesel industry has come under attack by some environmental associations, and subsidies for biofuel production have been condemned by some governments. Yet, biodiesel may represent a truly competitive alternative to diesel fuel, for which fuel tax exemption and subsidies to energetic crops are needed. Biodiesel must increase its popularity among social movements and governments to constitute a valid alternative of energy source. In this sense, the use of nonedible oils to produce biodiesel is proposed in the present review. Moreover, the compromise of noninterference between land for energetic and food purposes must be addressed. Concerning technical issues, it is important to consider a transesterification optimization, which is missing or incomplete for too many vegetable oils already tested. In most cases, a common...

Activity levels of oxidative stress-related enzymes in the root apoplast during the interaction of WR315 (resistant) and JG62 (susceptible) chickpeas (Cicer arietinum L.) with the highly virulent race 5 of Fusarium oxysporum f. sp. ciceris were compared. Because this fungus develops asymptomatic infections in the chickpea root cortex in both susceptible and resistant plants, but only intrudes into the root xylem in the susceptible variety, the interactions were compared at three specific stages during disease development in JG62: (i) before symptom development (10 days after inoculation); (ii) at the time of appearance of the first disease symptoms (15-17 days after inoculation) and (iii) when all plants had developed disease symptoms (20-22 days after inoculation). Diamine oxidase (DAO), ascorbate peroxidase (APX), glutathione reductase (GR), guaiacol-dependent peroxidase and superoxide dismutase (SOD), but not catalase (CAT), were found in the apoplast of chickpea roots. In terms of APX activity, infection by the pathogen caused a different response in the incompatible compared to the compatible plant. In the case of GR, SOD and DAO activities, the pathogen caused the same response, but it developed earlier (i.e. GR and SOD) or to higher levels (i.e. DAO) in the incompatible interaction. Expression of apx, cat, sod, lipoxygenase (lox) and actin genes was also analysed in infected roots. Infection by F. oxysporum f. sp. ciceris race 5 only caused a significant change in the root expression of lox and actin genes. This up-regulation was earlier (lox) or higher (actin) in the incompatible than in the compatible interaction. Thus, changes in oxidative metabolism differ in compatible and incompatible interactions in Fusarium wilt of chickpea.

Dam-recorded information was checked out in a selection scheme of dairy goats in South-eastern Spain. A total of 388 dam-offspring verifications were molecularly achieved using nine microsatellite markers. Five of these markers (SR-CRSP-1, SR-CRSP-5, SR-CRSP-8, SR-CRSP-9 and INRA011c) were chosen from the literature according to their polymorphic information content (PIC). Three markers (ChirUCO2, ChirUCO4 and ChirUCO5) were cloned in the same nucleus, and the remaining one was the ETH10 bovine microsatellite. The number of alleles observed were 10, 7, 7, 14, 21, 8, 7, 7, and 7 for the markers SR-CRSP-1, SR-CRSP-5, SR-CRSP-8, SR-CRSP-9, INRA011c, ChirUCO2, ChirUCO4, ChirUCO5 and ETH10, respectively. The global exclusion probability accomplished was 0.9991. On the other hand, among the 388 verifications carried out, 71.9% (279) resulted compatible, while 16.2% (63) were clearly incompatible. Most of the incompatibilities (84.1%) were due to less than four markers. These results suggest that a high percentage of dam-registered information in the selection nucleus could be erroneous, due to the archaic system used for animal identification and for data transfer. Thus an automated method of identification and data reporting should be considered to reduce the high level of errors encountered.

Nine new full-length cDNA sequences for γ-gliadins were generated from total mRNA from bread and durum wheat. The length of the open reading frames (ORFs) ranged from 720 to 936 bp, which corresponds to putative proteins of 239–311 amino acid residues, respectively. All nine sequences, reconstructed from phylogenetic data showed sequences similar to the γ-gliadins registered at GenBank. All known γ-gliadin sequences fall into four major groups in the tree constructed by the neighbour-joining method. Primer pairs were designed to discriminate between these four γ-gliadin groups during grain development in bread wheat using quantitative real time polymerase chain reaction (QRT-PCR). Expression profiles of two lines of bread wheat showed higher levels of gene transcription for γ-gliadins from group II, whereas γ-gliadins from group I were expressed at the lowest level. Thus, the γ-gliadins from group II are expressed four- and 10-fold higher in ‘Perico’ and ‘Anza’, respectively, than the other γ-gliadins. The results reported may help to understand the order of synthesis and deposition of γ-gliadins in the grain of developing bread wheat. In addition, the differences found in the expression levels among the four groups could be used to select genotypes with specific profiles of γ-gliadins. These findings will also facilitate the up- and down-regulation of γ-gliadin gene expression in transgenic wheat.

One of the main limitations of cereal breeding is the lack of genetic variability within cultivated crops. Hordeum chilense is a wild relative of Hordeum vulgare, which has been successfully used in the synthesis of amphiploids by crossing with Triticum spp. Among the agronomic traits of these new amphiploids, the allelic variation in the endosperm storage proteins and their influence on breadmaking and malting quality are of special interest. B-hordeins are sulfur rich prolamins, which account for 70-80% of the total hordein fraction in barley. In this work, rapid amplification of cDNA ends by PCR (RACE-PCR) has been used for the cloning of the full-length open reading frame (ORF) of six sequences of B3-hordeins from two lines of H. chilense. Two consensus sequences of 813 and 822 bp for the H1 and H7 lines, respectively, were determined by alignment of all the sequences generated. Between both lines, differences involving single base changes, which could correspond to single nucleotide polymorphisms (SNP), insertions and deletions were observed. Of these differences, only six out of the 13 within the ORF caused a change of amino acid. Two insertions/deletions of 9 and 12 bp were also observed between both lines. The derived amino acid sequences showed a similar structure to the B-hordeins from cultivated barley and other prolamins. The repetitive region is based on the repetition of the motif PQQPFPQQ. The copy number of the B3-hordeins was estimated as a minimum of nine and five copies for the H1 and H7 lines, respectively. The expression profile of the B-hordeins through the developing endosperm is also described in this work. This study of the storage proteins of H. chilense is a useful contribution to the knowledge of the genetic diversity available in wild relatives of cultivated barley. In addition, the origin of the different prolamins can be better understood with an in-depth knowledge of its wild equivalent.

A selection of 51 barley Sequence-Tagged Sites (STSs) were studied for their utility in Hordeum chilense. They included four primer sets from wheat origin and six primer sets from oat origin. Forty-four primer pairs amplified H. chilense products consistently. Five primer pairs were suitable for studying the introgression of H. chilense in wheat because they amplified H. chilense products of distinct size. Six of the STSs showed polymorphism between different H. chilense accessions. The results showed that barley STSs could be useful for the genetic characterization of H. chilense, tritordeums and derived introgression lines.

Hordeum chilense is a wild relative of H. vulgare, cultivated barley, that has been successfully used in the synthesis of amphiploids by crossing with Triticum spp. These amphiploids-named generically x Tritordeum-have been tested under field conditions, and one of them, the hexaploid tritordeum obtained following chromosome doubling of the hybrid H. chilense x T. turgidum, shows traits of interest inherited from the barley parent. Of great interest is the allelic variation observed in the endosperm storage proteins and their influence on the breadmaking and malting quality of tritordeum. We report here two mRNA (cDNA) sequences for a gamma-3 hordein from two accession lines of H. chilense, H1 and H7, and their characterization by quantitative real time (QRT)-PCR in the developing endosperm. Sequences were obtained by rapid amplification of cDNA ends and "edge-to-edge" amplification of open reading frames from cDNA of H. chilense. Eight putative single nucleotide polymorphisms and one codon insertion were identified in the sequences of the H1 and H7 gamma-3 hordeins. The deduced amino acid sequences showed similar features to that of the gamma-3 hordein and gamma-gliadins from barley and wheat, respectively. While the repetitive motif (PQQQPF) is similar to that of the gamma-3 hordein from H. vulgare, there are 19 motif repeats in H. vulgare, whereas H. chilense shows 15 tandem repeats. The transcription of the genes encoding for the gamma-3 hordein were monitored by QRT-PCR: in both lines maximum transcription occurred 12 days after flowering.

When developing SCARs by sequencing RAPD markers useful for olive variety identification, one RAPD sequence of 407 bp has been identified that shows significant DNA homology with more than 160 retrotransposon-like sequences. A generally coherent phylogenetic tree has been constructed based on the homologous retrotransposon-like sequences, reflecting genetic distances in 35 species belonging to eight kingdoms. The implications of this finding in the development of SCAR markers are discussed. In addition, specific oligonucleotide primers have been developed to amplify the DNA region and have a practical application as an internal amplification control in SCAR-based multiplexed PCRs. To our knowledge, this is the first report of a retrotransposon-like element in olive., Financial support from the CICYT andINIA, Spain (Projects OLI96–2184 and CAO98–001-C3–2), isalso acknowledged.

The common fig tree (Ficus carica L.) is a Mediterranean crop with problematic cultivar identification. The recovery and conservation of possible local varieties for ecological production requires the previous genetic characterization of the available germplasm. In this context, 42 lines corresponding to 12 local varieties and two caprifigs, in addition to 15 reference samples have been fingerprinted using 21 SSR markers. A total of 77 alleles were revealed, detecting a useful level of genetic variability within the local germplasm pools. UPGMA clustering analysis has revealed the genetic structure and relationships among the local and reference germplasm. Eleven of the local varieties could be identified and defined as obtained clusters, showing that SSR analysis is an efficient method to evaluate the Andalusian fig tree diversity for on-farm conservation.

Hordeum chilense is a potential source of useful genes for wheat breeding. The use of this wild species to increase genetic variation in wheat will be greatly facilitated by marker-assisted introgression. In recent years, the search for the most suitable DNA marker system for tagging H. chilense genomic regions in a wheat background has lead to the development of RAPD and SCAR markers for this species. RAPDs represent an easy way of quickly generating suitable introgression markers, but their use is limited in heterogeneous wheat genetic backgrounds. SCARs are more specific assays, suitable for automatation or multiplexing. Direct sequencing of RAPD products is a cost-effective approach that reduces labour and costs for SCAR development. The use of SSR and STS primers originally developed for wheat and barley are additional sources of genetic markers. Practical applications of the different marker approaches for obtaining derived introgression products are described., Financial support from the Spanish ‘Ministerio de Ciencia y Tecnología’ ('Ramón y Cajal' Program and project AGL2002‐04079‐C02‐02 from DGI) is gratefully acknowledged.

The transferability of microsatellite markers developed for olive cultivars ( Olea europaea L.) has been tested and confirmed in the Olea complex. Thirty two genotypes, belonging to different taxa of the genus Olea, have been analyzed with four olive SSRs. Positive amplifications at all loci were obtained in 13 taxa (at least one accession per species). Sixty seven different alleles have been detected at the four loci analyzed. Polymorphic products have been observed at the inter- and intra-species level. Some SSR loci have shown multiple amplification products in some species. The high number of unique alleles has allowed the unambiguous discrimination of most accessions. Similarity coefficients and relationships among the Olea taxa have been calculated based on SSR amplification results. The reliability of SSRs as markers for intra-species variability evaluation has been confirmed while their use to explore relationships at the inter-species level is discussed, being dependent on the locus analyzed.

[EN] A range of single tagged site (STS), simple sequence repeat (SSR), and sequence-characterized amplified region (SCAR) markers were screened for their utility in detecting Hordeum vulgare and H. chilense chromosomes in a wheat background. PCR conditions were optimized for specific amplification of the targeted sequences and to avoid cross-species amplification. Two H. vulgare derived STSs, six H. vulgare derived SSRs, and nine H. chilense derived SCARs were usable for the detection of five H. vulgare and three H. chilense chromosomes by direct ethidium bromide staining of the PCR products in test tubes, avoiding the more costly and time-consuming DNA electrophoresis step. The practical application of the method is illustrated by the identification of a monotelosomic substitution of H. vulgare chromosome 6HS in tritordeum and a monosomic addition of H. chilense chromosome 6Hch in durum wheat.Key words: GISH, Hordeum chilense, Hordeum vulgare, marker-assisted selection, SSR, Tritordeum., [FR] Un ensemble de marquers de type STS (« single-tagged-site »), microsatellite (SSR) et SCAR (« sequence-characterized amplified region ») ont été examinés quant à leur utilité potentielle pour la détection de chromosomes duHordeum vulgareet duH. chilensedans un fond génétique du blé. Les conditions PCR ont été optimisées pourl’amplification spécifique des séquences ciblées et pour éviter l’amplification croisée entre espèces. Deux marqueursSTS dérivés duH. vulgare, six microsatellites dérivés duH. vulgareet neuf SCAR dérivés duH. chilensepermettaientde détecter cinq chromosomes duH. vulgareet trois chromosomes duH. chilensepar simple détection des ampliconsau bromure d’éthidium dans des tubes réactionnels, évitant ainsi l’étape plus longue et coûteuse de l’électrophorèse.L’intérêt pratique de cette méthode est illustré via l’identification d’une substitution monotélosomique du chromosome6HS duH. vulgarechez une lignée issue d’un croisement blé-orge (tritordeum) ainsi que par l’identification d’une ad-dition monosomique du chromosome 6HchduH. chilensechez le blé durum., Financial support from the Minesterio de Ciencia y Tecnologia, Spain, the Comisión Interministerial de Ciencia yTecnologia (CICYT), Spain (project numbers AGF98-0945-CO2-01 and AGF99-1036-CO2-02), and the Spanish–British Acciones Integradas program (concerted action HB1998-0199) is acknowledged.

RAPD markers generated by mixtures of two different 10-mer primers were developed for eight different olive cultivars used as parental lines in olive-breeding programs. Two RAPD bands were converted into dominant SCAR markers by direct sequencing of the RAPD products, avoiding the costly and time-consuming cloning step. The SCARs generated have maintained the original RAPD polymorphism among the cultivars and segregated according to Mendelian inheritance. Preliminary results suggest the use of the SCAR SCOeMS-2 for the marker-assisted selection of the high flesh/stone ratio. This strategy provides a rapid method for the characterization of RAPD markers and for the development of PCR-based markers with applications in olive mapping, paternal testing and germplasm characterization. The use of these markers in multiplexed PCRs, and the direct ethidium bromide detection of the PCR products in the test tube, facilitate their efficient and reliable breeding applications., Financial support from the CICYT andINIA, Spain (Projects OLI96-2184 and CAO98-001-C3-2), is acknowledged.

The first genetic map of the wild South Ameri- can barley species Hordeum chilense is presented. The map, based on an F2 population of 114 plants, contains 123 markers, including 82 RAPDs, 13 SSRs, 16 RFLPs, four SCARs, two seed storage proteins and two STS markers. The map spans 694 cM with an average distance of 5.7 cM between markers. Six additional SSRs and seven additional SCARs which were not polymorphic were assigned to chromosomes using wheat/H. chilense addition lines. Polymorphisms were revealed by 50% of the RAPD amplifications, 13% of wheat and barley SSR primers, and 78% of the Gramineae RFLP anchor probes. The utility of SSR and RFLP probes from other Gramineae species shows the usefulness of a comparative approach as a source of markers and for aligning the genetic map of H. chilense with other species. This also indicates that the overall structure of the H. chilense linkage groups is probably similar to that of the B and D genomes of wheat and the H genome of barley. Applications of the map for tritordeum and wheat breeding are discussed., Financial support from the Spanish-Britishconcerted action, HB1998–0199, and from the CICYT, Spain (Project number AGF98–0945-CO2–01), is gratefully acknowl-edged.

A survey of Gramineae markers was carried out with the aim of developing cost-effective methods for the molecular analysis of Miscanthus species. Ten out of twenty Gramineae RFLP probes from ”anchor” sets hybridized well to Miscanthus DNA while all 15 maize probes tested cross-hybridized successfully, showing similar patterns in both species. Cross-taxa amplification of maize microsatellite primers was then tested. This showed that 57 out of 76 (75%) give highly reproducible amplification with Miscanthus DNA. Amplification products differed in size from those in maize but there was no bias toward higher or lower molecular weights. Microsatellite polymorphism produced by 17 primer pairs was studied in detail in a panel of 11 Miscanthus clones belonging to the species Miscanthus sinensis, Miscanthus sacchariflorus, Miscanthus ×giganteus and Miscanthus condensatus. Intra- and inter-specific length polymorphisms were frequent between the tested Miscanthus clones with length polymorphisms being found for all primer pairs, detecting 3–22 alleles. Polymorphism information content (PIC) values for microsatellites ranged from 0.48 to 0.94 with an average of 0.83. Species-specific amplicons were produced by two microsatellites. Genetic similarity coefficients of the Miscanthus clones ranged from 0.35 to 0.92, with an average of 0.57. Five polymorphisms were studied in a segregating population, where they showed Mendelian inheritance. In addition, two microsatellite markers mapping 1.3-cM apart on maize chromosome 7 were linked in Miscanthus at an estimated distance of 8 cM, suggesting collinearity. The high transferability of microsatellite markers from maize will enhance the power and resolution of genome analysis in Miscanthus., Financial support from the EU (Project num-ber FAIR-CT98–3571) and from the Spanish-British concerted ac-tion, HB1998–0199, are gratefully acknowledged.

RAPD markers were developed for octoploid×Tritordeum (amphiploid Hordeum chilense×Triticum aestivum) and its parents. Addition lines were used to identify specific RAPD markers for the Hordeum chilense chromosomes detectable in a wheat background. Twelve RAPD fragments have been cloned, sequenced and converted into STS markers. Eleven of these STSs have maintained both the chromosome specificity and the possibility of detection in a wheat background. The use of these markers in multiplexed PCRs facilitates both the efficient and reliable screening of new addition lines as well as the monitoring of introgression of H. chilense in bread and durum wheat., Th first author acknowledges the Consejo Su-perior de Investigaciones Cientificas''for the award of a fellowship .Financial support from the CICYT, Madrid, Spain (Project num-bers AGF95-0964-C02-01 and AGF98-0945-C02-01) is also ac-knowledged.

We have isolated and characterized 47 ultraviolet light-induced hprt mutants from a simian virus 40-transformed excision-repair-deficient xeroderma pigmentosum cell line (complementation group A). Twenty-one independent mutations were found, of which the majority were point mutations. Eleven of these were identified as base changes, nine of which could be attributed to ultraviolet damage on the transcribed DNA strand. Both transitions and transversions were found among the single base changes. A large proportion of the mutations ( 13 21 ) resulted in aberrant splicing of the hprt gene, suggesting that the target size for mutations resulting in aberrant splicing must be quite large. A small number of spontaneous mutations were identified, most of which were large deletions. Our data provide a spectrum for the intrinsic mutations resulting from ultraviolet damage in human cells in the absence of repair.

Homozygous AdhF AdhF and Adhs Adhs lines, collected from the same D. melanogaster winery polymorphic population, were submitted to stress by environmental ethanol. The responses of each of the FF, FS and SS genotypes, for both control and selected lines were analysed in normal and ethanol supplemented medium, in relation to three fitness components: egg-to-adult viability, developmental time and ethanol utilisation. Ethanol selection has resulted in a higher ethanol tolerance, a shorter developmental time and a better ability to use ethanol as food, showing the AdhF AdhF genotype to be that with the highest relative fitness.

The L-arabinose resistance test with Salmonella typhimurium (Ara test) is a forward mutation assay which selects a single phenotypic change (from L-arabinose sensitivity to L-arabinose resistance) in a unique tester strain (an araD mutant). The present study examined the ability of the Ara test to identify as mutagens different classes of chemical carcinogens, including six with detection problems in most bacterial assays. A noncarcinogen related in chemical structure to the selected carcinogen was included whenever possible. A total of 25 organic compounds was assayed by means of a standard mutagenesis procedure: the plate-incorporation test in the presence (if required) of 10% S9 from rat liver induced with Aroclor 1254. Chemicals giving negative or inconclusive results were then retested using other common in vitro mutagenesis conditions. The Ara test detected as mutagens all but one (Urethane) of the 15 established carcinogens and six out of the nine chemicals with questionable or nonassayed carcinogenicity. The compound mutagenic at the lowest dose was ethionine (0.57 nmol), one of the carcinogens undetected by the popular histidine reverse mutation assay (Ames test) or by the SOS chromotest. Actually, only benzo(a)pyrene was found mutagenic at a dose (0.77 nmol) close to that of ethionine. The data reported in this paper suggest that the Ara forward mutation test is less prone than the His reverse mutation test or the SOS chromotest to misclassify as negative carcinogenic compounds. Consequently, it should be considered as an alternative to other bacterial assays in general, massive, and primary screening for genotoxic carcinogens.

The forward mutation assay to L-arabinose resistance (Ara test) in Salmonella typhimurium detected a Spanish red table wine (Rioja) as a direct-acting mutagen. The best mutagenic response was obtained by preincubating strain BA13 with the wine sample in the presence of sodium phosphate buffer and in the absence of any external metabolic activation. In fact, the S9 mixture abolished most of the mutagenic activity of red wine in the Ara test. Such an inactivating capacity seems to be independent of microsomal enzymes and mediated through some kind of heat-stable component(s) in the S9 fraction. Both regular wine (directly from the bottle) and lyophilized wine were strong mutagens in the Ara test, inducing 4914 and 2739 AraR mutants/ml. Both pKM101 and delta uvrB were critical factors in the detection of the mutagenicity of wine, exhibiting a synergic effect in strain BA13. The mutagenicity of red wine was somehow pH-dependent, increasing with the pH value of the preincubation mixture. In comparison with the Ara test, the His reverse mutation assay (Ames test) was much less sensitive to the mutagenicity of lyophilized red wine, TA102 being the most (448 His+/ml) and TA98 the least (38 His+/ml) sensitive strain. TA100, TA104 and TA97 manifested intermediate mutagenicities to red wine. Previous reports have identified strain TA98 as the His strain most sensitive to the apolar fraction (e.g. XAD-2-bound) of red wine. Based on these results we propose that TA98 mainly detects glycosides of mutagenic flavonols present in red wine (quercetin, rutin, etc.), which do not constitute the major direct-acting mutagens detected with the Ara test.(ABSTRACT TRUNCATED AT 250 WORDS)

We report the development of microsatellites or simple sequence repeats (SSRs) in the olive tree (Olea europaea L.). Forty three positive clones obtained by the screening of a GA-enriched genomic library were sequenced and primers were designed for 13 microsatellite loci. Five primer pairs amplified polymorphic products of the expected size range. SSR polymorphism was explored in a set of 46 olive cultivars. A total of 26 alleles were detected for the five loci. Heterozygosity ranged from 0.46 to 0.71. Ninety one per cent of the cultivars had unique multilocus genotypes. Microsatellite segregation was studied in a complex population from a cross between the commercial cultivars ’Leccino’ and ’Dolce Agogia’.