Selective Inhibition of Vascular Endothelial Growth Factor–mediated Angiogenesis by Cyclosporin A: Roles of the Nuclear Factor of Activated T Cells and Cyclooxygenase 2

Cyclosporin A (CsA) is an immunosuppressive drug that inhibits the activity of transcription factors of the nuclear factor of activated T cells (NFAT) family, interfering with the induction of cytokines and other inducible genes required for the immune response. Here we show that CsA inhibits migration of primary endothelial cells and angiogenesis induced by vascular endothelial growth factor (VEGF); this effect appears to be mediated through the inhibition of cyclooxygenase (Cox)-2, the transcription of which is activated by VEGF in primary endothelial cells. Consistent with this, we show that the induction of Cox-2 gene expression by VEGF requires NFAT activation. Most important, the CsA-mediated inhibition of angiogenesis both in vitro and in vivo was comparable to the Cox-2 inhibitor NS-398, and reversed by prostaglandin E2. Furthermore, the in vivo corneal angiogenesis induced by VEGF, but not by basic fibroblast growth factor, was selectively inhibited in mice treated with CsA systemically. These findings involve NFAT in the regulation of Cox-2 in endothelial cells, point to a role for this transcription factor in angiogenesis, and may provide a novel mechanism underlying the beneficial effects of CsA in angiogenesis-related diseases such as rheumatoid arthritis and psoriasis.This work was supported by grant PM99-0116 from Ministerio de Educación y Cultura (MEC-DGES) of Spain (to J.M. Redondo) and grants FEDER 1FD97-0514-CO2-01 and FEDER FD97-0275 from MEC-DGES and the European Community to J.M. Redondo and M. Fresno, respectively. G.L. Hernández was supported by grants from Consejo Superior de Investigaciones Científicas y Tecnológicas (CONICET) of Argentina and Comunidad Autónoma de Madrid grant 8.3/0024/2000, and M. Fresno by grant PM97-0130, O. Volpert by American Heart Association grant AHA SDG 0030023N, and S. Martínez-Martínez by grant 8.3/19/1998 from the Comunidad Autónoma de Madrid. The Centro de Biología Molecular "Severo Ochoa" is supported by a grant from the Fundación Ramón ArecesPeer reviewe

Agromyces italicus sp. nov., Agromyces humatus sp. nov. and Agromyces lapidis sp. nov., isolated from Roman catacombs

A polyphasic study was carried out to clarify the taxonomic positions of three Gram-positive isolates from the Catacombs of Domitilla, Rome (Italy). 16S rRNA gene sequence comparisons placed these strains within the genus Agromyces. The morphological and chemotaxonomic characteristics of these isolates were consistent wiih the description of the genus Agromyces. The three isolates could be readily distinguished from one another and from representatives of all Agromyces species with validly published names by a broad range of phenotypic characteristics and DNA-DNA relatedness studies. Therefore, these isolates are proposed to represent three novel species of the genus Agromyces, Agromyces italicus sp. nov. (type strain CD1T=HKI 0325T=DSM 16388T=NCIMB 14011T), Agromyces humatus sp. nov. (type strain CD5T=HKI 0327T=DSM 16389T=NCIMB 14012T) and Agromyces lapidis sp. nov. (type strain CD55T =HKI 0324T=DSM 16390T=NCIMB 14013T). © 2005 IUMS.V. J. and L. L. are grateful to fellowships from the Spanish Ministry of Education and Science (MEC) I3P programme and J. M. G. to an MEC contract from the ‘Ramo´n y Cajal’ programme. This study was supported by project CATS (EVK4-CT2000-00028) and MEC project BTE2002-04492-C02-01.Peer Reviewe

Selective Inhibition of Vascular Endothelial Growth Factor–Mediated Angiogenesis by Cyclosporin a: Roles of the Nuclear Factor of Activated T Cells and Cyclooxygenase 2

Cyclosporin A (CsA) is an immunosuppressive drug that inhibits the activity of transcription factors of the nuclear factor of activated T cells (NFAT) family, interfering with the induction of cytokines and other inducible genes required for the immune response. Here we show that CsA inhibits migration of primary endothelial cells and angiogenesis induced by vascular endothelial growth factor (VEGF); this effect appears to be mediated through the inhibition of cyclooxygenase (Cox)-2, the transcription of which is activated by VEGF in primary endothelial cells. Consistent with this, we show that the induction of Cox-2 gene expression by VEGF requires NFAT activation. Most important, the CsA-mediated inhibition of angiogenesis both in vitro and in vivo was comparable to the Cox-2 inhibitor NS-398, and reversed by prostaglandin E2. Furthermore, the in vivo corneal angiogenesis induced by VEGF, but not by basic fibroblast growth factor, was selectively inhibited in mice treated with CsA systemically. These findings involve NFAT in the regulation of Cox-2 in endothelial cells, point to a role for this transcription factor in angiogenesis, and may provide a novel mechanism underlying the beneficial effects of CsA in angiogenesis-related diseases such as rheumatoid arthritis and psoriasis

Phyllobacterium catacumbae sp. nov., a member of the order 'Rhizobiales' isolated from Roman catacombs

Two strains were isolated from tuff, a volcanic rock that forms the walls of the Roman Catacombs of Saint Callixtus in Rome, Italy. A polyphasic approach using nutritional and physiological tests, reactions to antibiotics, fatty acid profiles, DNA base ratios, DNA-DNA reassociation and 16S rRNA gene sequence comparisons showed that the two isolates belong to a novel species within the genus Phyllobacterium. The species Phyllobacterium catacumbae sp. nov. is proposed. The type strain is CSC19T (=CECT 5680T=LMG 22520T).V. J. and L. L. received fellowships from the Spanish Ministry of Education and Science (MEC), I3P programme and J. M. G. is supported by an MEC contract from the ‘Ramón y Cajal’ program. This study was supported by EC project EVK4-CT2000-00028 and MEC project BTE2002-04492-C02-01.Peer Reviewe

Influence of potential pulses amplitude sequence in a voltammetric electronic tongue (VET) applied to assess antioxidant capacity in aliso

[EN] Four signals configurations were studied, two of them built by small increases of potential and two with bigger increments. The highest current values were obtained when pulses with bigger change of potential were used although the best results were shown by the pulse sequence which included an intermediate pulse before the relevant pulse. A mathematical model based on trolox pattern was developed to predict antioxidant capacity of aliso, employing information obtained from all the electrodes, although model validation could be done only employing the information from gold electrode.Fuentes-Pérez, EM.; Alcañiz Fillol, M.; Contat-Rodrigo, L.; Baldeon-Chamorro, E.; Barat Baviera, JM.; Grau Meló, R. (2017). Influence of potential pulses amplitude sequence in a voltammetric electronic tongue (VET) applied to assess antioxidant capacity in aliso. Food Chemistry. 224:233-241. doi:10.1016/j.foodchem.2016.12.076S23324122

La restauración de la villa romana de La Ontavia (Terrinches, Ciudad Real)

En el mes de julio de 2010 se iniciaron las labores de conservación y restauración en el yacimiento romano de La Ontavia (Terrinches, Ciudad Real). Durante cuatro meses se han simultaneado estos trabajos con la excavación arqueológica, en la que han colaborado once alumnos de la Escuela Superior de Conservación y Restauración de Bienes Culturales de la Comunidad de Madrid, bajo la dirección del equipo técnico, realizando prácticas para completar su formación. La intervención llevada a cabo ha estado enfocada a minimizar el efecto de los agentes de deterioro a los que se halla expuesto el yacimiento, que se encuentra al aire libre. El presente artículo tiene como finalidad la difusión de los trabajos de conservación y restauración realizados durante esta campaña, prestando especial atención a la metodología y a los materiales utilizadosIn July 2010 conservation and restoration work began at the Roman site of La Ontavia (Terrinches, Ciudad Real). For four months, the work was carried out alongside the archaeological excavation, involving eleven students from the Madrid School of Cultural Asset Conservation and Restoration, doing work experience under the guidance of a technical team to complete their training. The intervention focused on minimising the effect of the deterioration factors that this open-air site has been exposed to. This article aims to provide a report of the conservation and restoration work carried out in this campaign, paying special attention to the methodology and materials use

Museum Genomics Confirms that the Lord Howe Island Stick Insect Survived Extinction

The Lord Howe Island stick insect, Dryococelus australis, was once common on the island but was driven to extinction after the arrival of ship rats in the early 20th century [1, 2]. It was thought to be extinct for decades, until a tiny population of similar-looking stick insects was discovered 20 km away, on the islet of Ball’s Pyramid, in 2001 [2]. Individuals from this population are currently being reared in Australia and elsewhere in the world, with the eventual goal of recolonizing Lord Howe Island [3]. Recent surveys of the wild population on Ball’s Pyramid suggest that it is among the world’s rarest species. However, there are significant morphological differences between Ball’s Pyramid and museum specimens, and there has never been a genetic confirmation of the rediscovered population’s species identity. Because Dryococelus is monotypic, there are also no known extant relatives for comparison. Using shotgun genomic data from the Ball’s Pyramid population, we assembled a draft genome and the complete mitochondrial genome. We found that the genome is massive, over 4 Gb in size, and is most likely hexaploid. We re-sequenced mitochondrial genomes from historic museum specimens collected on Lord Howe Island before the extinction event. Sequence divergence between the two populations is less than 1% and is within the range of intraspecific differences between the museum specimens, suggesting that they are conspecific and that D. australis has successfully evaded extinction so far. This work highlights the importance of museum collections for taxonomic validation in the context of ongoing conservation efforts

Coralsnake venomics: Analyses of venom gland transcriptomes and proteomes of six Brazilian taxa

Venom gland transcriptomes and proteomes of six Micrurus taxa (M. corallinus, M. lemniscatus carvalhoi, M. lemniscatus lemniscatus, M. paraensis, M. spixii spixii, and M. surinamensis) were investigated, providing the most comprehensive, quantitative data on Micrurus venom composition to date, and more than tripling the number of Micrurus venom protein sequences previously available. The six venomes differ dramatically. All are dominated by 2–6 toxin classes that account for 91–99% of the toxin transcripts. The M. s. spixii venome is compositionally the simplest. In it, three-finger toxins (3FTxs) and phospholipases A2 (PLA2s) comprise >99% of the toxin transcripts, which include only four additional toxin families at levels ≥0.1%. Micrurus l. lemniscatus venom is the most complex, with at least 17 toxin families. However, in each venome, multiple structural subclasses of 3FTXs and PLA2s are present. These almost certainly differ in pharmacology as well. All venoms also contain phospholipase B and vascular endothelial growth factors. Minor components (0.1–2.0%) are found in all venoms except that of M. s. spixii. Other toxin families are present in all six venoms at trace levels (<0.005%). Minor and trace venom components differ in each venom. Numerous novel toxin chemistries include 3FTxs with previously unknown 8- and 10-cysteine arrangements, resulting in new 3D structures and target specificities. 9-cysteine toxins raise the possibility of covalent, homodimeric 3FTxs or heterodimeric toxins with unknown pharmacologies. Probable muscarinic sequences may be reptile-specific homologs that promote hypotension via vascular mAChRs. The first complete sequences are presented for 3FTxs putatively responsible for liberating glutamate from rat brain synaptosomes. Micrurus C-type lectin-like proteins may have 6–9 cysteine residues and may be monomers, or homo- or heterodimers of unknown pharmacology. Novel KSPIs, 3× longer than any seen previously, appear to have arisen in three species by gene duplication and fusion. Four species have transcripts homologous to the nociceptive toxin, (MitTx) α-subunit, but all six species had homologs to the β-subunit. The first non-neurotoxic, non-catalytic elapid phospholipase A2s are reported. All are probably myonecrotic. Phylogenetic analysis indicates that the six taxa diverged 15–35 million years ago and that they split from their last common ancestor with Old World elapines nearly 55 million years ago. Given their early diversification, many cryptic micrurine taxa are anticipated

Coralsnake Venomics: Analyses of Venom Gland Transcriptomes and Proteomes of Six Brazilian Taxa

Venom gland transcriptomes and proteomes of six Micrurus taxa (M. corallinus, M. lemniscatus carvalhoi, M. lemniscatus lemniscatus, M. paraensis, M. spixii spixii, and M. surinamensis) were investigated, providing the most comprehensive, quantitative data on Micrurus venom composition to date, and more than tripling the number of Micrurus venom protein sequences previously available. The six venomes differ dramatically. All are dominated by 2–6 toxin classes that account for 91–99% of the toxin transcripts. The M. s. spixii venome is compositionally the simplest. In it, three-finger toxins (3FTxs) and phospholipases A2 (PLA2s) comprise >99% of the toxin transcripts, which include only four additional toxin families at levels ≥0.1%. Micrurus l. lemniscatus venom is the most complex, with at least 17 toxin families. However, in each venome, multiple structural subclasses of 3FTXs and PLA2s are present. These almost certainly differ in pharmacology as well. All venoms also contain phospholipase B and vascular endothelial growth factors. Minor components (0.1–2.0%) are found in all venoms except that of M. s. spixii. Other toxin families are present in all six venoms at trace levels (<0.005%). Minor and trace venom components differ in each venom. Numerous novel toxin chemistries include 3FTxs with previously unknown 8- and 10-cysteine arrangements, resulting in new 3D structures and target specificities. 9-cysteine toxins raise the possibility of covalent, homodimeric 3FTxs or heterodimeric toxins with unknown pharmacologies. Probable muscarinic sequences may be reptile-specific homologs that promote hypotension via vascular mAChRs. The first complete sequences are presented for 3FTxs putatively responsible for liberating glutamate from rat brain synaptosomes. Micrurus C-type lectin-like proteins may have 6–9 cysteine residues and may be monomers, or homo- or heterodimers of unknown pharmacology. Novel KSPIs, 3× longer than any seen previously, appear to have arisen in three species by gene duplication and fusion. Four species have transcripts homologous to the nociceptive toxin, (MitTx) α-subunit, but all six species had homologs to the β-subunit. The first non-neurotoxic, non-catalytic elapid phospholipase A2s are reported. All are probably myonecrotic. Phylogenetic analysis indicates that the six taxa diverged 15–35 million years ago and that they split from their last common ancestor with Old World elapines nearly 55 million years ago. Given their early diversification, many cryptic micrurine taxa are anticipated