Evaluation of the CTSL2 gene as a candidate gene for alopecia X in Pomeranians and Keeshonden

Alopecia X is a noninflammatory, progressive, bilateral symmetric alopecia in dogs. The disease is mainly found in Nordic breeds. The breed predisposition and a strong familial accumulation suggest a hereditary background. We analyzed the cathepsin L2 gene (CTSL2) as a candidate for alopecia X. The comparative sequencing of 14 affected and 18 control animals revealed ten polymorphisms; however, none of these polymorphisms affected the coding sequence. Haplotype analysis did not reveal an association of one particular CTSL2 haplotype with the disease phenotype; therefore, we conclude that the CTSL2 gene is probably not the causative gene for alopecia X.

Speciation with gene-flow in eastern North American red oaks (Quercus section Lobatae)

As foundational species, oaks (Quercus : Fagaceae) support the activities of both humans and wildlife. However, many oaks in North America are declining, a crisis exacerbated by the previous disappearance of other hard mast-producing trees. In addition, the economic demands placed on this drought-tolerant group may intensify if climate change extirpates other, relatively mesophytic species. Genetic tools can help address these management challenges. To this end, we developed a suite of 27 microsatellite markers, of which 22 are derived from expressed sequence tags (ESTs). Many of these markers bear significant homology to known genes and may be able to directly assay functional genetic variation. Markers obtained from enriched microsatellite libraries, on the other hand, are typically located in heterochromatic regions and should reflect demographic processes. Considered jointly, genic and genomic microsatellites can elucidate patterns of gene-flow and natural selection, which are fundamental to both an organism's evolutionary ecology and conservation biology. To this end, we employed the developed markers in an FST-based genome scan to detect the signature of divergent selection among the red oaks (Quercus section Lobatae). Three candidate genes with putative roles in stress responses demonstrated patterns of diversity consistent with adaptation to heterogeneous selective pressures. These genes may be important in both local genetic adaptation within species and divergence among them. Next, we used an isolation-with-migration model to quantify levels of gene-flow among four red oaks species during speciation. Both speciation in allopatry and speciation with gene-flow were found to be major drivers of red oak biodiversity. Loci playing a key role in speciation are also likely to be ecologically important within species

Studies on the {\it in vivo} function of a phosphatidylinositol transfer protein from {\it Saccharomyces cerevisiae}

Phosphatidylinositol transfer proteins (PI-TP's) catalyze the transfer of phosphatidylinositol and phosphatidylcholine between membranes in vitro. However the in vivo function of these proteins is unknown. In this thesis we have used a combined biochemical and genetic approach to determine the importance of PI-TP in vivo. An oligonucleotide based on the amino terminal sequence of the PI-TP from Saccharomyces cerevisiae, was used to screen a yeast genomic library for the gene encoding PI-TP (PIT1 gene). Yeast strains transformed with the positive clones showed overproduction of transfer activities and transfer protein in the 100,000 x g supernatants. The 5$\sp\prime$ terminus of the PIT1 gene correlates with the predicted codons for residues 3-30 of the determined protein sequence. Tetrad analysis of a heterozygous diploid (PIT1/pit1::LEU2) revealed that the PIT1 gene is essential for cell growth. Non-viable spores could be rescued by transformation of the above diploid prior to sporulation, with a plasmid borne copy of the wild type gene. Sequencing of the entire PIT1 gene has revealed that the PIT1 gene is identical to the SEC14 gene. The sec14 ts mutant which exhibits conditional defects at the Golgi stage of protein secretion, is also temperature sensitive for PI-TP activity in vitro. These findings represent the first instance in which a physiological function has been assigned to any phospholipid transfer protein. ^

Genomic cloning and expression of three murine UDP-galactose: beta-N-acetylglucosamine beta1,3-galactosyltransferase genes.

Based on the detection of expressed sequence tags that are similar to known galactosyltransferase sequences, we have isolated three novel UDP-galactose:beta-N-acetylglucosamine beta1, 3-galactosyltransferase (beta3GalT) genes from a mouse genomic library. The three genes, named beta3GalT-I, -II, and -III, encode type II transmembrane proteins of 326, 422, and 331 amino acids, respectively. The three proteins constitute a distinct subfamily as they do not share any sequence identity with other eucaryotic galactosyltransferases. Also, the entire protein-coding region of the three beta3GalT genes was contained in a single exon, which contrasts with the genomic organization of the beta1,4- and alpha1, 3-galactosyltransferase genes. The three beta3GalT genes were mainly expressed in brain tissue. The expression of the full-length murine genes as recombinant baculoviruses in insect cells revealed that the beta3GalT enzymes share the same acceptor specificity for beta-linked GlcNAc, although they differ in their Km for this acceptor and the donor UDP-Gal. The identification of beta3GalT genes emphasizes the structural diversity present in the galactosyltransferase gene family.

Phylogenetic position of Riemerella anatipestifer based on 16S rRNA gene sequences

Riemerella anatipestifer, the causative agent of septicemia anserum exsudativa (also called new duckling disease), belongs to the family Flavobacteriaceae of gram-negative bacteria. We determined the DNA sequences of the rrs genes encoding the 16S rRNAs of four R. anatipestifer strains by directly sequencing PCR-amplified rrs genes. A sequence similarity analysis confirmed the phylogenetic position of R. anatipestifer in the family Flavobacteriaceae in rRNA superfamily V and allowed fine mapping of R. anatipestifer on a separate rRNA branch comprising the most closely related species, Bergeyella zoohelcum, as well as Chryseobacterium balustinum, Chryseobacterium indologenes, and Chryseobacterium gleum. The sequences of the rrs genes of the four R. anatipestifer strains varied between 0.5 and 1.0%, but all of the strains occupied the same position on the phylogenetic tree. In general, differences in rrs genes were observed among R. anatipestifer strains, even within a given serotype, as shown by restriction fragment length polymorphism of PCR-amplified rrs genes.

Phylogenetic positions of Clostridium chauvoei and Clostridium septicum based on 16S rRNA gene sequences

The sequences of the 16S rRNA genes (rrs genes) of Clostridium chauvoei, the causative agent of blackleg in cattle, and the phenotypically related organism Clostridium septicum were determined. After amplification of 1,507-bp PCR fragments from the corresponding rrs genes, the sequences were determined in a single round of sequencing by using conserved region primers. A sequence similarity analysis of the sequences revealed the close phylogenetic relationship of C. chauvoei and C. septicum in Clostridium cluster I (M. D. Collins, P. A. Lawson, A. Willems, J. J. Cordoba, J. Fernandez-Garayzabal, P. Garcia, J. Cai, H. Hippe, and J. A. E. Farrow, Int. J. Syst. Bacteriol. 44:812-826, 1994), which includes Clostridium carnis, Clostridium perfringens, Clostridium botulinum, and Clostridium tetani. We found that 99.3% of the nucleotides in the genes of C. chauvoei and C. septicum are identical.

The Molecular Characterization of a Predicted Glycosyltransferase from Lycopersicon esculentum

The synthesis of the plant cell wall is very complex, and understanding how this process occurs will lead to many benefits for future research and industries dependent upon cell walls for their products. The recent discovery of the functions of AtMUR3 and AtGT18 in Arabidopsis thaliana as xyloglucan galactosyltransferases has led to the identification of many more putative glycosyltransferases in the Arabidopsis genome. Due to the structural differences between the xyloglucans of Arabidopsis and solanaceous plants, we decided to search for putative arabinosyltransferases in the Solanaceae. Solanaceous xyloglucan is substituted by one to two arabinosyl residues at the second xylose position, and sometimes contains an arabinose at the first xylose position. In contrast, Arabidopsis xyloglucan does not contain arabinose, and is substituted by galactose at the second and third xylose position. Furthermore, the second galactose residue in Arabidopsis xyloglucan is usually fucosylated, a modification not found in solanaceous plants. Searching the database of expressed sequence tags (dbEST), we identified many likely glycosyltransferases in solanaceous plants, including tomato (Lycopersicon esculentum). AtMUR3 and AtGT18 search queries resulted in the identification of three putative glycosyltransferases in L. esculentum, which were tentatively designated LeGT1, Le1GT18, and Le2GT18. Based on phylogenetic considerations, Le2GT18 was thought to be a putative arabinosyltransferase. The gene was transformed into atmur3-3 and atgt18 mutant plants, and the resulting plants will be screened for homozygous plants with the inserted gene. The homozygous T2 plants can then be screened for changes in the composition of their cell walls. Because Le2GT18 is thought to be an arabinosyltransferase, the levels of arabinose may be increased in the xyloglucan fraction of the cell wall. If so, further testing can be performed to reveal the true function of Le2GT18.

Preliminary results and documentation of sediment cores CRP2 and CRP-2A from the Ross Sea off Cape Roberts, Antarctica

The site for CRP-2, 14 km east of Cape Roberts (77.006°S; 163.719°E), was selected to overlap the early Miocene strata cored in nearby CRP-1, and to sample deeper into the east-dipping strata near the western margin ofe he Victoria Land Basin to investigate Palaeogene climatic and tectonic history. CRP-2 was cored from 5 to 57 mbsf (metres below the sea floor) (core recovery 91 %), with a deviation resulting in CRP-2A being cored at the same site. CRP-2A reached down to 624mbsf (recovery 95%), and to strata with an age of c. 33-35 Ma. Drilling took place from 16 October to 25 November 1998, on 2.0-2.2 m of sea ice and through 178 m of water. Core fractures and other physical properties, such as sonic velocity, density and magnetic susceptibility, were measured throughout the core. Down-hole logs for these and other properties were run from 63 to 167 mbsf and subsequently from 200 to 623 mbsf, although density and velocity data could be obtained only to 440 mbsf because of hole collapse. Sonic velocity averages c. 2.0 km S-1 for the upper part of the hole, but there is an sharp increase to c. 3.0 km s-1 and also a slight angular unconformity, at 306 mbsf, corresponding most likely to the early/late Oligocene boundary (c. 28-30 Ma). Velocity then increases irregularly to around 3.6 km s-1 at the bottom of the hole, which is estimated to lie 120 m above the V4/V5 boundary. The higher velocities below 306 mbsf probably reflect more extensive carbonate and common pyrite cementation, in patches, nodules, bedding-parallel masses and as vein infills. Dip of the strata also increases down-hole from 3° in the upper 300 in to over 10° at the bottom. Temperature gradient is 21° k-1. Over 2 000 fractures were logged through the hole. Borehole televiewer imagery was obtained for the interval from 200 to 440 mbsf to orient the fractures for stress field analysis. Lithostratigraphical descriptions on a scale of 1:20 are presented for the full length of the core, along with core box images, as a 200 page supplement to this issue. The hole initially passed through a layer of muddy gravel to 5.5 mbsf (Lithological Sub-Unit or LSU 1.1), and then into a Quaternary diatom-bearing clast-rich diamicton to 21 mbsf (LSU 2. l), with an interval of alternating compact diamicton and loose sand, and containing a rich Pliocene foraminiferal fauna, to 27 mbsf (LSU 2.2). The unit beneath this (LSU 3.1) has similar physical properties (sonic velocity, porosity, magnetic susceptibility) and includes diamictites of similar character to those of LSU 2.1 and 2.2, but an early Miocene (c. 19 Ma) diatom assemblage at 28 mbsf (top of LSU 3.1) shows that this sub-unit is part of the older section. The strata beneath 27 mbsf, primary target for the project, extend from early Miocene to perhaps latest Eocene age, and are largely cyclic glacimarine nearshore to offshore sediments. They are described as 41 lithological sub-units and interpreted in terms of 12 recurrent lithofacies. These are 1) mudstone, 2) inter-stratified mudstone and sandstone, 3) muddy very fine to coarse sandstone, 4) well-sorted stratified fine sandstone, 5) moderately to well-sorted, medium-grained sandstone, 6) stratified diamictite, 7) massive diamictite, 8) rhythmically inter-stratified sandstone and mudstone, 9) clast-supported conglomerate, 10) matrix-supported conglomerate, 11) mudstone breccia and 12) volcaniclastic sediment. Sequence stratigraphical analysis has identified 22 unconformity-bounded depositional sequences in pre- Pliocene strata. They typically comprise a four-part architecture involving, in ascending order, 1) a sharp-based coarse-grained unit (Facies 6,7,9 or 10), 2) a fining-upward succession of sandstones (Facies 3 and 4), 3) a mudstone interval (Facies l), in some cases coarsening upward to muddy sandstones (Facies 3), and 4) a sharp-based sandstone dominated succession (mainly Facies 4). The cyclicity recorded by the strata is interpreted in terms of a glacier ice margin retreating and advancing from land to the west, and of rises and falls in sea level. Analysis of sequence periodicity awaits afirmer chronology. However, apreliminary spectral analysis of magnetic susceptibility for a deepwater mudstone within one of the sequences (from 339 to 347 mbsf) reveals ratios between hierarchical levels that are similar to those of the three Milankovitch orbital forcing periodicities. The strata contain a wide range of fossils, the most abundant being marine diatoms. These commonly form up to 5% of the sediment, though in places the core is barren (notably between 300 and 412 mbsf). Fifty samples out of 250 reviewed were studied in detail. The assemblages define ten biostratigraphical zones, some of them based on local or as yet undescribed forms. The assemblages are neritic, and largely planktonic, suggesting that the sea floor was mostly below the photic zone throughout deposition of the corcd sequence. Calcareous nannofossils, representing incursions of ocean surface waters, are much less common (72 out of 183 samples examined) and restricted to mudstone intervals a few tens of metres thick, but are important for dating. Foraminifera are also sparse (73 out of 135 samples) and represented only by calcareous benthic species. Changing assemblages indicate a shift from inshore environments in the early Oligocenc to outer shelf in the late Oligocenc, returning to inshore in the early Miocene. Marine palynomorplis yielded large numbers of well-preserved forms from most of the 116 samples examined. The new in situ assemblagc found last year in CRP-1 is extended down into the late Oligocene and a further new assemblage is found in the early Oligoccnc. Many taxa are new, and cannot us yet contribute to an improved understanding of chronology or ecology. Marine invertebrate macrofossils, mostly molluscs and serpulid tubes, are scattered throughout the core. Preservation is good in mudstones but poor in other lithologies. Climate on land is reflected in the content of terrestrial palynomorphs, which are extremely scarce down to c. 300 mbsf. Some forms are reworked, and others represent a low growing sparse tundra with at least one species of Nothofagus. Beneath this level, a significantly greater diversity and abundance suggests a milder climate and a low diversity woody vegetation in the early Oligocene, but still far short of the richness found in known Eocene strata of the region. Sedimentary facies in the oldest strata also suggest a milder climate in the oldest strata cored, with indications of substantial glacial melt-water discharges, but are typical of a coldcr climate in late Oligocene and early Miocene times. Clast analyses from diamictites reveal weak to random fabrics, suggesting either lack of ice-contact deposition or post-depositional modification, but periods when ice grounded at the drill site are inferred from thin zones of in-situ brecciated rock and soft-sediment folding. These are more common above c. 300 mbsf, perhaps reflecting more extensive glacial advances during deposition of those strata. Erosion of the adjacent Transantarctic Mountains through Jurassic basalt and dolerite-intruded Beacon strata into basement rocks beneath is recorded by petrographical studies of clast and sand grain assemblages. Core below 310 mbsf contains a dominance of fine-grained Jurassic dolerite and basalt fragments along with Beacon-derived coal debris and rounded quartz grains, whereas the strata above this level have a much higher proportion of basement derived granitoids, implying that the large areas of the adjacent mountains had been eroded to basement by the end of the early Oligocene. There is little indication of rift-related volcanism below 310 mbsf. Above this, however, basaltic and trachytic tephras are common, especially from 280 to 200 mbsf, from 150 to 46 mbsf, and in Pliocene LSU 2.2 from 21 to 27 mbsf. The largest volcanic eruptions generated layers of coarse (up to 1 cm) trachytic pumice lapilli between 97 and 114 mbsf. The thickest of these (1.2 m at 112 mbsf) may have produced an eruptive column extending tens of km into the stratosphere. A source within a few tens of km of the drill site is considered most likely. Present age estimates for the pre-Pliocene sequence are based mainly on biostratigraphy (using mainly marine diatoms and to a lesser extent calcareous nannofossils), with the age of the tephra from 112 to 114 mbsf (21.44k0.05 Ma from 84 crystals by Ar-Ar) as a key reference point. Although there are varied and well-preserved microfossil assemblages through most of the sequence (notably of diatoms and marine palynomorphs), they comprise largely taxa either known only locally or as yet undescribed. In addition, sequence stratigraphical analysis and features in the core itself indicate numerous disconformities. The present estimate from diatom assemblages is that the interval from 27 to 130 mbsf is early Miocene in age (c. 19 to 23.5 Ma), consistent with the Ar-Ar age from 112 to 114 mbsf. Diatom assemblages also indicate that the late Oligocene epoch extends from c. 130 to 307 mbsf, which is supported by late Oligocene nannofossils from 130 to 185 mbsf. Strata from 307 to 412 mbsf have no age-diagnostic assemblages, but below this early Oligocene diatoms and nannofossils have been recovered. A nannoflora at the bottom of the hole is consistent with an earliest Oligocene or latest Eocene age. Magnetostratigraphical studies based on about 1000 samples, 700 of which have so far undergone demagnetisation treatment, have provided a polarity stratigraphy of 12 pre-Pliocene magnetozones. Samples above 270 mbsf are of consistently high quality. Below this, magnetic behaviour is more variable. A preliminary age-depth plot using the Magnetic Polarity Time Scale (MPTS) and constrained by biostratigraphical data suggests that episodes of relatively rapid sedimentation took place at CRP-2 during Oligocene times (c. 100 m/My), but that more than half of the record was lost in a few major and many minor disconformities. Age estimates from Sr isotopes in shell debris and further tephra dating are expected to lead to a better comparison with the MPTS. CRP-2/2A has recorded a history of subsidence of the Victoria Land Basin margin that is similar to that found in CIROS-170 km to the south, reflecting stability in both basin and the adjacent mountains in late Cenozoic times, but with slow net accumulation in the middle Cenozoic. The climatic indicators from both drill holes show a similar correspondence, indicating polar conditions for the Quaternary but with sub-polar conditions in the early Miocene-late Oligocene and indications of warmer conditions still in the early Oligocene. Correlation between the CRP-2A core and seismic records shows that seismic units V3 and V4, both widespread in the Victoria Land Basin, represent a period of fluctuating ice margins and glacimarine sedimentation. The next drill hole, CRP-3, is expected to core deep into V5 and extend this record of climate and tectonics still further back in time.

Geochemistry, clay mineralogy and grain size distribution in various sediment cores drilled during ODP Leg 119 on the Antarctic continental shelf off Prydz Bay

Drilling was undertaken at five sites (739-743) on ODP Leg 119 on a transect across the continental shelf of Prydz Bay, East Antarctica, to elucidate the long-term glacial history of the area and to examine the importance of the area with respect to the development of the East Antarctic ice sheet as a whole. In addition to providing a record of glaciation spanning 36 m.y. or more, Leg 119 has provided information concerning the development of a continental margin under the prolonged influence of a major ice sheet. This has allowed the development of a sedimentary model that may be applicable not only to other parts of the Antarctic continental margin, but also to northern high-latitude continental shelves. The cored glacial sedimentary record in Prydz Bay consists of three major sequences, dominated by diamictite: 1. An upper flat-lying sequence that ranges in thickness from a few meters in inner and western Prydz Bay to nearly 250 m in the outer or eastern parts of the bay. The uppermost few meters consist of Holocene diatom ooze and diatomaceous mud with a minor ice-rafted component overlying diamicton and diamictite of late Miocene to Quaternary age. The diamictite is mainly massive, but stratified varieties and minor mudstone and diatomite also occur. 2. An upper prograding sequence cored at Sites 739 and 743, unconformly below the flat-lying sequence. This consists of a relatively steep (4° inclination) prograding wedge with a number of discrete sedimentary packages. At Sites 739 and 743 the sequence is dominated by massive and stratified diamictite, some of which shows evidence of slumping and minor debris flowage. 3. A lower, more gently inclined, prograding sequence lies unconformably below the flat-lying sequence at Site 742 and the upper prograding sequence at Site 739. This extends to the base of both sites, to 316 and 487 mbsf, respectively. It is dominated by massive, relatively clast-poor diamictite which is kaolinite-rich, light in color, and contains sporadic carbonate-cemented layers. The lower part of Site 742 includes well-stratified diamictites and very poorly sorted mudstones. The base of this site has indications of large-scale soft-sediment deformation and probably represents proximity to the base of the glacial sequence. Facies analysis of the Prydz Bay glacial sequence indicates a range of depositional environments. Massive diamictite is interpreted largely as waterlain till, deposited close to the grounding line of a floating glacier margin, although basal till and debris flow facies are also present. Weakly stratified diamictite is interpreted as having formed close to or under the floating ice margin and influenced by the input of marine diatomaceous sediment (proximal glaciomarine setting). Well-stratified diamictite has a stronger marine input, being more diatom-rich, and probably represents a proximal-distal glaciomarine sediment with the glaciogenic component being supplied by icebergs. Other facies include a variety of mudstones and diatom-rich sediments of marine origin, in which an ice-rafted component is still significant. None of the recovered sediments are devoid of a glacial influence. The overall depositional setting of the prograding sequence is one in which the grounded ice margin is situated close to the shelf edge. Progradation was achieved primarily by deposition of waterlain till. The flat-lying sequence illustrates a complex sequence of advances and retreats across the outer part of the shelf, with intermittent phases of ice loading and erosion. The glacial chronology is based largely on diatom stratigraphy, which has limited resolution. It appears that ice reached the paleoshelf break by earliest Oligocene, suggesting full-scale development of the East Antarctic ice sheet by that time. The ice sheet probably dominated the continental margin for much of Oligocene to middle Miocene time. Retreat, but not total withdrawal of the ice sheet, took place in late Miocene to mid-Pliocene time. The late Pliocene to Pleistocene was characterized by further advances across, and progradation of, the continental shelf. Holocene time has been characterized by reduced glacial conditions and a limited influence of glacial processes on sedimentation.

¿El museo como forma de "conocer la Historia"? El público interpreta la inmigración

El trabajo analiza cómo la exposición presentada en el Museo Nacional de la Inmigración fue aprehendida e interpretada por su público visitante. Dependiente de la Dirección Nacional de Migraciones, el Museo funcionó entre los años 2001 y 2009 en el edificio del antiguo Hotel de Inmigrantes (en Retiro). Para este estudio utilizamos más de 700 mensajes registrados en el Libro de Visitas del Museo, que fueron clasificados en nueve categorías, de acuerdo a los aspectos comunes que expresaban. A lo largo del análisis, el concepto de memoria se volvió fundamental para comprender los sentimientos y deseos expresados por el público que visitó la institución. Dentro de los temas que aparecieron en los mensajes, pudimos verificar la importancia que le atribuían los visitantes al Estado argentino por la construcción del Hotel, la recepción de los inmigrantes y, actualmente, por la preservación de su memoria y patrimonio; también se evidenció un agradecimiento a los inmigrantes, cuya "fuerza" y "esperanza" contribuyeron al desarrollo del país

¿El museo como forma de "conocer la Historia"? El público interpreta la inmigración

El trabajo analiza cómo la exposición presentada en el Museo Nacional de la Inmigración fue aprehendida e interpretada por su público visitante. Dependiente de la Dirección Nacional de Migraciones, el Museo funcionó entre los años 2001 y 2009 en el edificio del antiguo Hotel de Inmigrantes (en Retiro). Para este estudio utilizamos más de 700 mensajes registrados en el Libro de Visitas del Museo, que fueron clasificados en nueve categorías, de acuerdo a los aspectos comunes que expresaban. A lo largo del análisis, el concepto de memoria se volvió fundamental para comprender los sentimientos y deseos expresados por el público que visitó la institución. Dentro de los temas que aparecieron en los mensajes, pudimos verificar la importancia que le atribuían los visitantes al Estado argentino por la construcción del Hotel, la recepción de los inmigrantes y, actualmente, por la preservación de su memoria y patrimonio; también se evidenció un agradecimiento a los inmigrantes, cuya "fuerza" y "esperanza" contribuyeron al desarrollo del país

¿El museo como forma de "conocer la Historia"? El público interpreta la inmigración

El trabajo analiza cómo la exposición presentada en el Museo Nacional de la Inmigración fue aprehendida e interpretada por su público visitante. Dependiente de la Dirección Nacional de Migraciones, el Museo funcionó entre los años 2001 y 2009 en el edificio del antiguo Hotel de Inmigrantes (en Retiro). Para este estudio utilizamos más de 700 mensajes registrados en el Libro de Visitas del Museo, que fueron clasificados en nueve categorías, de acuerdo a los aspectos comunes que expresaban. A lo largo del análisis, el concepto de memoria se volvió fundamental para comprender los sentimientos y deseos expresados por el público que visitó la institución. Dentro de los temas que aparecieron en los mensajes, pudimos verificar la importancia que le atribuían los visitantes al Estado argentino por la construcción del Hotel, la recepción de los inmigrantes y, actualmente, por la preservación de su memoria y patrimonio; también se evidenció un agradecimiento a los inmigrantes, cuya "fuerza" y "esperanza" contribuyeron al desarrollo del país

Efecto de las cubiertas ajardinadas sobre el microclima urbano de verano

El cambio climático y sus efectos requieren con urgencia el desarrollo de estrategias capaces no solo de mitigar pero también permitir la adaptación de los sistemas afectados por este fenómeno a los cambios que están provocando a nivel mundial. Olas de calor más largas y frecuentes, inundaciones, y graves sequías aumentan la vulnerabilidad de la población, especialmente en asentamientos urbanos. Este fenómeno y sus soluciones potenciales han sido ampliamente estudiados en las últimas décadas desde diferentes perspectivas y escalas que analizan desde el fenómeno regional de isla de calor al aumento de la intensidad energética necesaria en los edificios para mantener las condiciones de confort en los escenarios de calentamiento que se predicen. Su comprensión requiere el entendimiento de este fenómeno y un profundo análisis de las estrategias que pueden corregirlo y adaptarse a él. En la búsqueda de soluciones a este problema, las estrategias que incorporan sistemas naturales tales como las cubiertas ajardinadas, las fachadas vegetadas y bosques urbanos, se presentan como opciones de diseño capaces de proporcionan múltiples servicios al ecosistema urbano y de regular y hacer frente a los efectos del cambio climático. Entre los servicios que aportan estos sistemas naturales se incluyen la gestión de agua de tormentas, el control del efecto isla de calor, la mejora de la calidad del aire y del agua, el aumento de la diversidad, y como consecuencia de todo lo anterior, la reducción de la huella ecológica de las ciudades. En la última década, se han desarrollado múltiples estudios para evaluar y cuantificar los servicios al ecosistema proporcionados por las infraestructuras verdes, y específicamente las cubiertas ajardinadas, sin embargo, determinados servicios como la capacidad de la regulación del microclima urbano no ha sido apenas estudiados. La mayor parte de la literatura en este campo la componen estudios relacionados con la capacidad de las cubiertas ajardinadas de reducir el efecto de la isla de calor, en una escala local, o acerca de la reducción de la demanda energética de refrigeración debida a la instalación de cubiertas ajardinadas en la escala de edificio. La escala intermedia entre estos dos ámbitos, la calle, desde su ámbito habitable cercano al suelo hasta el límite superior del cañón urbano que configura, no han sido objeto detallado de estudio por lo que es esta escala el objeto de esta tesis doctoral. Esta investigación tiene como objeto contribuir en este campo y aportar un mayor entendimiento a través de la cuantificación del impacto de las cubiertas ajardinadas sobre la temperatura y humedad en el cañón urbano en la escala de calle y con un especial foco en el nivel peatonal. El primer paso de esta investigación ha sido la definición del objeto de estudio a través del análisis y revisión de trabajos tanto teóricos como empíricos que investigan los efectos de cubiertas ajardinadas en el entorno construido, entendidas como una herramienta para la adaptación y mitigación del impacto del cambio climático en las ciudades. La literatura analizada, revela el gran potencial de los sistemas vegetales como herramientas para el diseño pasivo puesto que no solo son capaces de mejorar las condiciones climáticas y microclimaticas en las ciudades reduciendo su demanda energética, sino también la necesidad de mayor análisis en la escala de calle donde confluyen el clima, las superficies urbanas y materiales y vegetación. Este análisis requiere una metodología donde se integren la respuesta térmica de edificios, las variaciones en los patrones de viento y radiación, y la interacción con la vegetación, por lo que un análisis cuantitativo puede ayudar a definir las estrategias más efectivas para lograr espacios urbanos más habitables. En este contexto, el objetivo principal de esta investigación ha sido la evaluación cuantitativa del impacto de la cubierta ajardinada en el microclima urbano a escala de barrio en condiciones de verano en los climas mediterráneos continentales. Para el logro de este objetivo, se ha seguido un proceso que persigue identificar los modelos y herramientas de cálculo capaces de capturar el efecto de la cubierta ajardinada sobre el microclima, identificar los parámetros que potencian o limitan este efecto, y cuantificar las variaciones que microclima creado en el cañón urbano produce en el consumo de energía de los edificios que rodean éste espacio. La hipótesis principal detrás de esta investigación y donde los objetivos anteriores se basan es el siguiente: "una cubierta ajardinada instalada en edificios de mediana altura favorece el establecimiento de microclimas a nivel peatonal y reduce las temperaturas en el entorno urbano donde se encuentra”. Con el fin de verificar la hipótesis anterior y alcanzar los objetivos propuestos se ha seguido la siguiente metodología: • definición del alcance y limitaciones del análisis • Selección de las herramientas y modelos de análisis • análisis teórico de los parámetros que afectan el efecto de las cubiertas ajardinadas • análisis experimental; • modelización energética • conclusiones y futuras líneas de trabajo Dada la complejidad de los fenómenos que intervienen en la generación de unas determinadas condiciones microclimáticas, se ha limitado el objeto de este estudio a las variables de temperatura y humedad, y sólo se han tenido en cuenta los componentes bióticos y abióticos del sistema, que incluyen la morfología, características superficiales del entorno estudiado, así como los elementos vegetales. Los componentes antrópicos no se han incluido en este análisis. La búsqueda de herramientas adecuadas para cumplir con los objetivos de este análisis ha concluido en la selección de ENVI-met v4 como el software más adecuado para esta investigación por su capacidad para representar los complejos fenómenos que caracterizan el microclima en cañones urbanos, en una escala temporal diaria y con unas escala local de vecindario. Esta herramienta supera el desafío que plantean los requisitos informáticos de un cálculo completo basado en elementos finitos realizados a través de herramientas de dinámica de fluidos computacional (CFD) que requieren una capacidad de cálculo computacional y tiempo privativos y en una escala dimensional y temporal limitada a esta capacidad computacional lo que no responde a los objetivos de esta investigación. ENVI-met 4 se basa es un modelo tridimensional del micro clima diseñado para simular las interacciones superficie-planta-aire en entornos urbanos. Basado en las ecuaciones fundamentales del equilibrio que representan, la conservación de masa, energía y momento. ENVI-met es un software predictivo, y como primer paso ha requerido la definición de las condiciones iniciales de contorno que se utilizan como punto de partida por el software para generar su propio perfil de temperatura y humedad diaria basada en la localización de la construcción, geometría, vegetación y las superficies de características físicas del entorno. La geometría de base utilizada para este primer análisis se ha basado en una estructura típica en cuanto al trazado urbano situada en Madrid que se ha simulado con una cubierta tradicional y una cubierta ajardinada en sus edificios. La estructura urbana seleccionada para este análisis comparativo es una red ortogonal con las calles principales orientadas este-oeste. El edificio típico que compone el vecindario se ha definido como “business as usual” (BAU) y se ha definido con una cubierta de baldosa de hormigón estándar, con un albedo 0.3, paredes con albedo 0.2 (construcción de muro de ladrillo típico) y cerramientos adiabáticos para evitar las posibles interferencias causadas por el intercambio térmico con el ambiente interior del edificio en los resultados del análisis. Para el caso de la cubierta ajardinada, se mantiene la misma geometría y características del edificio con excepción de la cobertura superficial de la azotea. Las baldosas de hormigón se han modificado con una cubierta ajardinada extensiva cubierta con plantas xerófilas, típicas en el clima de Madrid y caracterizado por su índice de densidad foliar, el “leaf area density” (LAD), que es la superficie total de superficie de hojas por unidad de volumen (m2/m3). El análisis se centra en los cañones urbanos entendidos como el espacio de calle comprendido entre los límites geométricos de la calle, verticales y horizontales, y el nivel superior de la cota urbana nivel de cubiertas. Los escenarios analizados se basan en la variación de la los principales parámetros que según la literatura analizada condicionan las variaciones microclimáticas en el ámbito urbano afectado por la vegetación, la velocidad del viento y el LAD de la azotea. Los resultados han sido registrados bajo condiciones de exposición solar diferentes. Las simulaciones fueron realizadas por los patrones de viento típico de verano, que para Madrid se caracterizan por vientos de componente suroeste que van desde 3 a 0 m/s. las simulaciones fueron realizadas para unas condiciones climáticas de referencia de 3, 2, 1 y 0 m/s a nivel superior del cañón urbano, como condición de contorno para el análisis. Los resultados calculados a 1,4 metros por encima del nivel del suelo, en el espacio habitado, mostraron que el efecto de la cubierta ajardinada era menor en condiciones de contorno con velocidades de viento más altas aunque en ningún caso el efecto de la cubierta verde sobre la temperatura del aire superó reducciones de temperatura de aire superiores a 1 º C. La humedad relativa no presentó variaciones significativas al comparar los diferentes escenarios. Las simulaciones realizadas para vientos con velocidad baja, entre 0 y 1 m/s mostraron que por debajo de 0.5 m/s la turbulencia del modelo aumentó drásticamente y se convirtió en el modelo inestable e incapaz de producir resultados fiables. Esto es debido al modelo de turbulencia en el software que no es válido para velocidades de viento bajas, lo que limita la capacidad de ENVI-met 4 para realizar simulaciones en estas condiciones de viento y es una de las principales conclusiones de este análisis en cuanto a la herramienta de simulación. También se comprobó el efecto de las densidades de la densidad de hoja (LAD) de los componentes vegetales en el modelo en la capa de aire inmediatamente superior a la cubierta, a 0,5 m sobre este nivel. Se compararon tres alternativas de densidad de hoja con la cubierta de baldosa de hormigón: el techo verde con LAD 0.3 (hierba típica o sedum), LAD 1.5 (plantas mixtas típicas) y LAD 2.5 (masa del árbol). Los resultados mostraron diferencias de temperatura muy relevante entre las diferentes alternativas de LAD analizadas. Los resultados muestran variaciones de temperatura que oscilan entre 3 y 5 º C al comparar el estándar de la azotea concreta con albedo 0, 3 con el techo con vegetación y vegetación densa, mostrando la importancia del LAD en la cuantificación de los efectos de las cubiertas vegetales en microclima circundante, lo que coincide con los datos reportados en la literatura existente y con los estudios empíricos analizados. Los resultados de los análisis teóricos han llegado a las siguientes conclusiones iniciales relacionadas con la herramienta de simulación y los resultados del modelo: En relación con la herramienta ENVI-met, se han observado limitaciones para el análisis. En primer lugar, la estructura rígida de la geometría, las bases de datos y el tamaño de la cuadrícula, limitan la escala y resolución de los análisis no permitiendo el desarrollo de grandes zonas urbanas. Por otro lado la estructura de ENVI-met permite el desarrollo de este tipo de simulación tan complejo dentro de tiempos razonables de cálculo y requerimientos computacionales convencionales. Otra limitación es el modelo de turbulencia del software, que no modela correctamente velocidades de viento bajas (entre 0 y 1 m/s), por debajo de 0,5 m/s el modelo da errores y no es estable, los resultados a estas velocidades no son fiables porque las turbulencias generadas por el modelo hacen imposible la extracción de patrones claros de viento y temperatura que permitan la comparación entre los escenarios de cubierta de hormigón y ajardinada. Además de las limitaciones anteriores, las bases de datos y parámetros de entrada en la versión pública del software están limitados y la complejidad de generar nuevos sistemas adaptándolos al edificio o modelo urbano que se quiera reproducir no es factible salvo en la versión profesional del software. Aparte de las limitaciones anteriores, los patrones de viento y perfiles de temperatura generados por ENVI-met concuerdan con análisis previos en los que se identificaban patrones de variación de viento y temperaturas en cañones urbanos con patrones de viento, relación de aspecto y dimensiones similares a los analizados en esta investigación. Por lo tanto, el software ha demostrado una buena capacidad para reproducir los patrones de viento en los cañones de la calle y capturar el efecto de enfriamiento producido por la cubierta verde en el cañón. En relación con el modelo, el resultado revela la influencia del viento, la radiación y el LAD en la temperatura del aire en cañones urbanos con relación de aspecto comprendida entre 0,5 y 1. Siendo el efecto de la cubierta verde más notable en cañones urbanos sombreados con relación de aspecto 1 y velocidades de viento en el nivel de “canopy” (por encima de la cubierta) de 1 m/s. En ningún caso las reducciones en la temperatura del aire excedieron 1 º C, y las variaciones en la humedad relativa no excedieron 1% entre los escenarios estudiados. Una vez que se han identificado los parámetros relevantes, que fueron principalmente la velocidad del viento y el LAD, se realizó un análisis experimental para comprobar los resultados obtenidos por el modelo. Para éste propósito se identificó una cubierta ajardinada de grandes dimensiones capaz de representar la escala urbana que es el objeto del estudio. El edificio usado para este fin fue el parking de la terminal 4 del aeropuerto internacional de Madrid. Aunque esto no es un área urbana estándar, la escala y la configuración del espacio alrededor del edificio fueron considerados aceptables para el análisis por su similitud con el contexto urbano objeto de estudio. El edificio tiene 800 x 200 m, y una altura 15 m. Está rodeado de vías de acceso pavimentadas con aceras conformando un cañón urbano limitado por el edificio del parking, la calle y el edificio de la terminal T4. El aparcamiento está cerrado con fachadas que configuran un espacio urbano de tipo cañón, con una relación de aspecto menor que 0,5. Esta geometría presenta patrones de viento y velocidad dentro del cañón que difieren ligeramente de los generados en el estudio teórico y se acercan más a los valores a nivel de canopo sobre la cubierta del edificio, pero que no han afectado a la tendencia general de los resultados obtenidos. El edificio cuenta con la cubierta ajardinada más grande en Europa, 12 Ha cubiertas por con una mezcla de hierbas y sedum y con un valor estimado de LAD de 1,5. Los edificios están rodeados por áreas plantadas en las aceras y árboles de sombra en las fachadas del edificio principal. El efecto de la cubierta ajardinada se evaluó mediante el control de temperaturas y humedad relativa en el cañón en un día típico de verano. La selección del día se hizo teniendo en cuenta las predicciones meteorológicas para que fuesen lo más semejantes a las condiciones óptimas para capturar el efecto de la cubierta vegetal sobre el microclima urbano identificadas en el modelo teórico. El 09 de julio de 2014 fue seleccionado para la campaña de medición porque las predicciones mostraban 1 m/s velocidad del viento y cielos despejados, condiciones muy similares a las condiciones climáticas bajo las que el efecto de la cubierta ajardinada era más notorio en el modelo teórico. Las mediciones se registraron cada hora entre las 9:00 y las 19:00 en 09 de julio de 2014. Temperatura, humedad relativa y velocidad del viento se registraron en 5 niveles diferentes, a 1.5, 4.5, 7.5, 11.5 y 16 m por encima del suelo y a 0,5 m de distancia de la fachada del edificio. Las mediciones fueron tomadas en tres escenarios diferentes, con exposición soleada, exposición la sombra y exposición influenciada por los árboles cercanos y suelo húmedo. Temperatura, humedad relativa y velocidad del viento se registraron con un equipo TESTO 410-2 con una resolución de 0,1 ºC para temperatura, 0,1 m/s en la velocidad del viento y el 0,1% de humedad relativa. Se registraron las temperaturas de la superficie de los edificios circundantes para evaluar su efecto sobre los registros usando una cámara infrarroja FLIR E4, con resolución de temperatura 0,15ºC. Distancia mínima a la superficie de 0,5 m y rango de las mediciones de Tª de - 20 º C y 250 º C. Los perfiles de temperatura extraídos de la medición in situ mostraron la influencia de la exposición solar en las variaciones de temperatura a lo largo del día, así como la influencia del calor irradiado por las superficies que habían sido expuestas a la radiación solar así como la influencia de las áreas de jardín alrededor del edificio. Después de que las medidas fueran tomadas, se llevaron a cabo las siguientes simulaciones para evaluar el impacto de la cubierta ajardinada en el microclima: a. estándar de la azotea: edificio T4 asumiendo un techo de tejas de hormigón con albedo 0.3. b. b. cubierta vegetal : T4 edificio asumiendo una extensa cubierta verde con valor bajo del LAD (0.5)-techo de sedum simple. c. c. cubierta vegetal: T4 edificio asumiendo una extensa cubierta verde con alta joven valor 1.5-mezcla de plantas d. d. cubierta ajardinada más vegetación nivel calle: el edificio T4 con LAD 1.5, incluyendo los árboles existentes a nivel de calle. Este escenario representa las condiciones actuales del edificio medido. El viento de referencia a nivel de cubierta se fijó en 1 m/s, coincidente con el registro de velocidad de viento en ese nivel durante la campaña de medición. Esta velocidad del viento se mantuvo constante durante toda la campaña. Bajo las condiciones anteriores, los resultados de los modelos muestran un efecto moderado de azoteas verdes en el microclima circundante que van desde 1 º a 2 º C, pero una contribución mayor cuando se combina con vegetación a nivel peatonal. En este caso las reducciones de temperatura alcanzan hasta 4 ºC. La humedad relativa sin embargo, no presenta apenas variación entre los escenarios con y sin cubierta ajardinada. Las temperaturas medidas in situ se compararon con resultados del modelo, mostrando una gran similitud en los perfiles definidos en ambos casos. Esto demuestra la buena capacidad de ENVI-met para reproducir el efecto de la cubierta ajardinada sobre el microclima y por tanto para el fin de esta investigación. Las diferencias más grandes se registraron en las áreas cercanas a las zonas superiores de las fachadas que estaban más expuestas a la radiación del sol y también el nivel del suelo, por la influencia de los pavimentos. Estas diferencias se pudieron causar por las características de los cerramientos en el modelo que estaban limitados por los datos disponibles en la base de datos de software, y que se diferencian con los del edificio real. Una observación importante derivada de este estudio es la contribución del suelo húmedo en el efecto de la cubierta ajardinada en la temperatura del aire. En el escenario de la cubierta ajardinada con los arboles existentes a pie de calle, el efecto del suelo húmedo contribuye a aumentar las reducciones de temperatura hasta 4.5ºC, potenciando el efecto combinado de la cubierta ajardinada y la vegetación a pie de calle. Se realizó un análisis final después de extraer el perfil horario de temperaturas en el cañón urbano influenciado por el efecto de las cubiertas ajardinadas y los árboles. Con esos perfiles modificados de temperatura y humedad se desarrolló un modelo energético en el edificio asumiendo un edificio cerrado y climatizado, con uso de oficinas, una temperatura de consigna de acuerdo al RITE de 26 ºC, y con los sistemas por defecto que establece el software para el cálculo de la demanda energética y que responden a ASHRAE 90.1. El software seleccionado para la simulación fue Design Builder, por su capacidad para generar simulaciones horarias y por ser una de las herramientas de simulación energética más reconocidas en el mercado. Los perfiles modificados de temperatura y humedad se insertaron en el año climático tipo y se condujo la simulación horaria para el día definido, el 9 de Julio. Para la simulación se dejaron por defecto los valores de conductancia térmica de los cerramientos y la eficiencia de los equipos de acuerdo a los valores que fija el estándar ASHRAE para la zona climática de Madrid, que es la 4. El resultado mostraba reducciones en el consumo de un día pico de hasta un 14% de reducción en las horas punta. La principal conclusión de éste estudio es la confirmación del potencial de las cubiertas ajardinadas como una estrategia para reducir la temperatura del aire y consumo de energía en los edificios, aunque este efecto puede ser limitado por la influencia de los vientos, la radiación y la especie seleccionada para el ajardinamiento, en especial de su LAD. Así mismo, en combinación con los bosques urbanos su efecto se potencia e incluso más si hay pavimentos húmedos o suelos porosos incluidos en la morfología del cañón urbano, convirtiéndose en una estrategia potencial para adaptar los ecosistemas urbanos el efecto aumento de temperatura derivado del cambio climático. En cuanto a la herramienta, ENVI-met se considera una buena opción para éste tipo de análisis dada su capacidad para reproducir de un modo muy cercano a la realidad el efecto de las cubiertas. Aparte de ser una herramienta validada en estudios anteriores, en el caso experimental se ha comprobado por medio de la comparación de las mediciones con los resultados del modelo. A su vez, los resultados y patrones de vientos generados en los cañones urbanos coinciden con otros estudios similares, concluyendo por tanto que es un software adecuado para el objeto de esta tesis doctoral. Como líneas de investigación futura, sería necesario entender el efecto de la cubierta ajardinada en el microclima urbano en diferentes zonas climáticas, así como un mayor estudio de otras variables que no se han observado en este análisis, como la temperatura media radiante y los indicadores de confort. Así mismo, la evaluación de otros parámetros que afectan el microclima urbano tales como variables geométricas y propiedades superficiales debería ser analizada en profundidad para tener un resultado que cubra todas las variables que afectan el microclima en el cañón urbano. ABSTRACT Climate Change is posing an urgency in the development of strategies able not only to mitigate but also adapt to the effects that this global problem is evidencing around the world. Heat waves, flooding and severe draughts increase the vulnerability of population, and this is especially critical in urban settlements. This has been extensively studied over the past decades, addressed from different perspectives and ranging from the regional heat island analysis to the building scale. Its understanding requires physical and dimensional analysis of this broad phenomenon and a deep analysis of the factors and the strategies which can offset it. In the search of solutions to this problem, green infrastructure elements such as green roofs, walls and urban forests arise as strategies able provide multiple regulating ecosystem services to the urban environment able to cope with climate change effects. This includes storm water management, heat island effect control, and improvement of air and water quality. Over the last decade, multiple studies have been developed to evaluate and quantify the ecosystem services provided by green roofs, however, specific regulating services addressing urban microclimate and their impact on the urban dwellers have not been widely quantified. This research tries to contribute to fill this gap and analyzes the effects of green roofs and urban forests on urban microclimate at pedestrian level, quantifying its potential for regulating ambient temperature in hot season in Mediterranean –continental climates. The study is divided into a sequence of analysis where the critical factors affecting the performance of the green roof system on the microclimate are identified and the effects of the green roof is tested in a real case study. The first step has been the definition of the object of study, through the analysis and review of theoretical and empirical papers that investigate the effects of covers landscaped in the built environment, in the context of its use as a tool for adaptation and mitigation of the impact of climate change on cities and urban development. This literature review, reveals the great potential of the plant systems as a tool for passive design capable of improving the climatic and microclimatic conditions in the cities, as well as its positive impact on the energy performance of buildings, but also the need for further analysis at the street scale where climate, urban surfaces and materials, and vegetation converge. This analysis requires a methodology where the thermal buildings response, the variations in the patterns of wind and the interaction of the vegetation are integrated, so a quantitative analysis can help to define the most effective strategies to achieve liveable urban spaces and collaterally, , the improvement of the surrounding buildings energy performance. In this specific scale research is needed and should be customized to every climate, urban condition and nature based strategy. In this context, the main objective for this research was the quantitative assessment of the Green roof impact on the urban microclimate at a neighbourhood scale in summer conditions in Mediterranean- continental climates. For the achievement of this main objective, the following secondary objectives have been set: • Identify the numerical models and calculation tools able to capture the effect of the roof garden on the microclimate. • Identify the enhancing or limiting parameter affecting this effect. • Quantification of the impact of the microclimate created on the energy consumption of buildings surrounding the street canyon analysed. The main hypothesis behind this research and where the above objectives are funded on is as follows: "An extensive roof installed in medium height buildings favours the establishment of microclimates at the pedestrian level and reduces the temperatures in the urban environment where they are located." For the purpose of verifying the above hypothesis and achieving the proposed objectives the following methodology has been followed: - Definition of hypothesis and objectives - Definition of the scope and limitations - Theoretical analysis of parameters affecting gren roof performance - Experimental analysis; - Energy modelling analyisis - Conclusions and future lines of work The search for suitable tools and models for meeting the objectives of this analysis has led to ENVI-met v4 as the most suitable software for this research. ENVI met is a three-dimensional micro-climate model designed to simulate the surface-plant-air interactions in urban environments. Based in the fundamental equations representing, mass, energy and momentum conservation, the software has the capacity of representing the complex phenomena characterizing the microclimate in urban canyons, overcoming the challenge posed by the computing requirements of a full calculus based on finite elements done via traditional computational fluid dynamics tools. Once the analysis tool has been defined, a first set of analysis has been developed to identify the main parameters affecting the green roof influence on the microclimate. In this analysis, two different scenarios are compared. A neighborhood with standard concrete tile roof and the same configuration substituting the concrete tile by an extensive green roof. Once the scenarios have been modeled, different iterations have been run to identify the influence of different wind patterns, solar exposure and roof vegetation type on the microclimate, since those are the most relevant variables affecting urban microclimates. These analysis have been run to check the conditions under which the effects of green roofs get significance. Since ENVI-met V4 is a predictive software, the first step has been the definition of the initial weather conditions which are then used as starting point by the software, which generates its own daily temperature and humidity profile based on the location of the building, geometry, vegetation and the surfaces physical characteristics. The base geometry used for this first analysis has been based on a typical urban layout structure located in Madrid, an orthogonal net with the main streets oriented East-West to ease the analysis of solar radiation in the different points of the model. This layout represents a typical urban neighborhood, with street canyons keeping an aspect ratio between 0.5 and 1 and high sky view factor to ensure correct sun access to the streets and buildings and work with typical wind flow patterns. Finally, the roof vegetation has been defined in terms of foliage density known as Leaf Area Density (LAD) and defined as the total one-sided leaf area per unit of layer volume. This index is the most relevant vegetation characteristic for the purpose of calculating the effect of vegetation on wind and solar radiation as well as the energy consumed during its metabolic processes. The building as usual (BAU) configuring the urban layout has been defined with standard concrete tile roofs, considering 0.3 albedo. Walls have been set with albedo 0.2 (typical brick wall construction) and adiabatic to avoid interference caused by thermal interchanges with the building indoor environment. For the proposed case, the same geometry and building characteristics have been kept. The only change is the roof surface coverage. The gravel on the roof has been changed with an extensive green roof covered with drought tolerant plants, typical in Madrid climate, and characterized by their LAD. The different scenarios analysed are based in the variation of the wind speed and the LAD of the roof. The results have been recorded under different sun exposure conditions. Simulations were run for the typical summer wind patterns, that for Madrid are characterized by South-west winds ranging from 3 to 0 m/s. Simulations were run for 3, 2, 1 and 0 m/s at urban canopy level. Results taken at 1.4 m above the ground showed that the green roof effect was lower with higher wind speeds and in any case the effect of the green roof on the air temperatures exceeded air temperature reductions higher than 1ºC. Relative humidity presented no variations when comparing the different scenarios. For the analysis at 0m/s, ENVI-met generated error and no results were obtained. Different simulations showed that under 0.5 m/s turbulence increased dramatically and the model became unstable and unable to produce reliable results. This is due to the turbulence model embedded in the software which is not valid for low wind speeds (below 1 m/s). The effect of the different foliage densities was also tested in the model. Three different alternatives were compared against the concrete roof: green roof with LAD 0.3 ( typical grass or sedum), 1.5 (typical mixed plants) and 2.5 (tree mass). The results showed very relevant temperature differences between the different LAD alternatives analyzed. Results show temperature variations ranging between 3 and 5 ºC when comparing the standard concrete roof with albedo 0, 3 with the vegetated roof and vegetated mass, showing the relevance of the LAD on the effects of green roofs on microclimate. This matches the data reported in existing literature and empirical studies and confirms the relevance of the LAD in the roof effect on the surrounding microclimate. The results of the theoretical analysis have reached the following initial conclusions related to both, the simulation tool and the model results: • In relation to the tool ENVI-met, some limitations for the analysis have been observed. In first place, the rigid structure of the geometry, the data bases and the grid size, limit the scale and resolution of the analysis not allowing the development of large urban areas. On the other hand the ENVI-met structure enables the development of this type of complex simulation within reasonable times and computational requirements for the purpose of this analysis. Additionally, the model is unable to run simulations at wind speeds lower than 0.5 m/s, and even at this speed, the results are not reliable because the turbulences generated by the model that made impossible to extract clear temperature differences between the concrete and green roof scenarios. Besides the above limitations, the wind patterns and temperature profiles generated by ENVImet are in agreement with previous analysis identifying wind patterns in urban canyons with similar characteristics and aspect ratio. Therefore the software has shown a good capacity for reproducing the wind effects in the street canyons and seems to capture the cooling effect produced by the green roof. • In relation to the model, the results reveals the influence of wind, radiation and LAD on air temperature in urban canyons with aspect ratio comprised between 0.5 and 1. Being the effect of the green roof more noticeable in shaded urban canyons with aspect ratio 1 and wind speeds of 1 m/s. In no case the reductions in air temperature exceeded 1ºC. Once the relevant parameters have been identified, mainly wind speed and LAD, an experimental analysis was conducted to test the results obtained by the model. For this purpose a large green roof was identified, able to represent the urban scale which is the object of the studio. The building identified for this purpose was the terminal 4, parking building of the international Madrid Airport. Even though this is not a standard urban area, the scale and configuration of the space around the building were deemed as acceptable for the analysis. The building is an 800x200 m, 15 m height parking building, surrounded by access paved paths and the terminal building. The parking is enclosed with facades that configure an urban canyon-like space, although the aspect ratio is lower than 0.5 and the wind patterns might differ from the theoretical model run. The building features the largest green roof in Europe, a 12 Ha extensive green roof populated with a mix of herbs and sedum with a LAD of 1.5. The buildings are surrounded by planted areas at the sidewalk and trees shading the main building facades. Green roof performance was evaluated by monitoring temperatures and relative humidity in the canyon in a typical summer day. The day selection was done taking into account meteorological predictions so the weather conditions on the measurement day were as close as possible as the optimal conditions identified in terms of green roof effects on the urban canyon. July 9th 2014 was selected for the measurement campaign because the predictions showed 1 m/s wind speed and sunny sky, which were very similar to the weather conditions where the effect of the green roof was most noticeable in the theory model. Measurements were registered hourly from 9:00am to 19:00 on July 9th 2014. Temperature, relative humidity and wind speed were recorded at 5 different levels, at 1.5, 4.5, 7.5, 11.5 and 16 m above ground and at 0.5 m distance from the building façade. Measurements were taken in three different scenarios, sunny exposure, shaded exposure, and shaded exposure influenced by nearby trees and moist soil. Temperature, relative humidity and wind speed were registered using a TESTO 410-2 anemometer, with 0.1ºC resolution for temperature, 0.1 m/s resolution for wind speed and 0.1 % for relative humidity. Surface temperatures were registered using an infrared camera FLIR E4, with temperature resolution 0.15ºC. Minimal distance to surface of 0.5 m and Tª measurements range from -20ºC and 250ºC. The temperature profiles measured on the site showed the influence of solar exposure on the temperature variations along the day, as well as the influence of the heat irradiated by the building surfaces which had been exposed to the sun radiation and those influenced by the moist soft areas around the building. After the measurements were taken, the following simulations were conducted to evaluate the impact of the green roof on the microclimate: a. Standard roof: T4 building assuming a concrete tile roof with albedo 0.3. b. Green roof: T4 building assuming an extensive green roof with low LAD value (0.5)-Simple Sedum roof. c. Green roof: T4 building assuming an extensive green roof with high LAD value 1.5- Lucerne and grasses d. Green roof plus street level vegetation: T4 Building, LAD 1.5 (Lucerne), including the existing trees at street level. This scenario represents the current conditions of the building. The urban canopy wind was set as 1 m/s, the wind speed register at that level during the measurement campaign. This wind speed remained constant over the whole campaign. Under the above conditions, the results of the models show a moderate effect of green roofs on the surrounding microclimate ranging from 1ºC to 2ºC, but a larger contribution when combining it with vegetation at pedestrian level, where 4ºC temperature reductions are reached. Relative humidity remained constant. Measured temperatures and relative humidity were compared to model results, showing a close match in the profiles defined in both cases and the good capacity of ENVI met to capture the impact of the green roof in this analysis. The largest differences were registered in the areas close to the top areas of the facades which were more exposed to sun radiation and also near to the soil level. These differences might be caused by differences between the materials properties included in the model (which were limited by the data available in the software database) and those in the real building. An important observation derived from this study is the contribution of moist soil to the green roof effect on air temperatures. In the green roof scenario with surrounding trees, the effect of the moist soil contributes to raise the temperature reductions at 4.5ºC. A final analysis was conducted after extracting the hourly temperature profile in the street canyon influenced by the effect of green roofs and trees. An energy model was run on the building assuming it was a conventional enclosed building. Energy demand reductions were registered in the building reaching up to 14% reductions at the peak hour. The main conclusion of this study is the potential of the green roofs as a strategy for reducing air temperatures and energy consumption in the buildings, although this effect can be limited by the influence of high speed winds. This effect can be enhanced its combination with urban forests and even more if soft moist pavements are included in the urban canyon morphology, becoming a potential strategy for adapting urban ecosystems to the increasing temperature effect derived from climate change.

A reversibly glycosylated polypeptide (RGP1) possibly involved in plant cell wall synthesis: Purification, gene cloning, and trans-Golgi localization

We purified from pea (Pisum sativum) tissue an ≈40 kDa reversibly glycosylated polypeptide (RGP1) that can be glycosylated by UDP-Glc, UDP-Xyl, or UDP-Gal, and isolated a cDNA encoding it, apparently derived from a single-copy gene (Rgp1). Its predicted translation product has 364 aminoacyl residues and molecular mass of 41.5 kDa. RGP1 appears to be a membrane-peripheral protein. Immunogold labeling localizes it specifically to trans-Golgi dictyosomal cisternae. Along with other evidence, this suggests that RGP1 is involved in synthesis of xyloglucan and possibly other hemicelluloses. Corn (Zea mays) contains a biochemically similar and structurally homologous RGP1, which has been thought (it now seems mistakenly) to function in starch synthesis. The expressed sequence database also reveals close homologs of pea Rgp1 in Arabidopsis and rice (Oryza sativa). Rice possesses, in addition, a distinct but homologous sequence (Rgp2). RGP1 provides a polypeptide marker for Golgi membranes that should be useful in plant membrane studies.

Structure, Properties, and Tissue Localization of Apoplastic α-Glucosidase in Crucifers1

Apoplastic α-glucosidases occur widely in plants but their function is unknown because appropriate substrates in the apoplast have not been identified. Arabidopsis contains at least three α-glucosidase genes; Aglu-1 and Aglu-3 are sequenced and Aglu-2 is known from six expressed sequence tags. Antibodies raised to a portion of Aglu-1 expressed in Escherichia coli recognize two proteins of 96 and 81 kD, respectively, in vegetative tissues of Arabidopsis, broccoli (Brassica oleracea L.), and mustard (Brassica napus L.). The acidic α-glucosidase activity from broccoli flower buds was purified using concanavalin A and ion-exchange chromatography. Two active fractions were resolved and both contained a 96-kD immunoreactive polypeptide. The N-terminal sequence from the 96-kD broccoli α-glucosidase indicated that it corresponds to the Arabidopsis Aglu-2 gene and that approximately 15 kD of the predicted N terminus was cleaved. The 81-kD protein was more abundant than the 96-kD protein, but it was not active with 4-methylumbelliferyl-α-d-glucopyranoside as the substrate and it did not bind to concanavalin A. In situ activity staining using 5-bromo-4-chloro-3-indolyl-α-d-glucopyranoside revealed that the acidic α-glucosidase activity is predominantly located in the outer cortex of broccoli stems and in vascular tissue, especially in leaf traces.