The origin of 15R-prostaglandins in the Caribbean coral Plexaura homomalla: Molecular cloning and expression of a novel cyclooxygenase

The highest concentrations of prostaglandins in nature are found in the Caribbean gorgonian Plexaura homomalla. Depending on its geographical location, this coral contains prostaglandins with typical mammalian stereochemistry (15S-hydroxy) or the unusual 15R-prostaglandins. Their metabolic origin has remained the subject of mechanistic speculations for three decades. Here, we report the structure of a type of cyclooxygenase (COX) that catalyzes transformation of arachidonic acid into 15R-prostaglandins. Using a homology-based reverse transcriptase–PCR strategy, we cloned a cDNA corresponding to a COX protein from the R variety of P. homomalla. The deduced peptide sequence shows 80% identity with the 15S-specific coral COX from the Arctic soft coral Gersemia fruticosa and ≈50% identity to mammalian COX-1 and COX-2. The predicted tertiary structure shows high homology with mammalian COX isozymes having all of the characteristic structural units and the amino acid residues important in catalysis. Some structural differences are apparent around the peroxidase active site, in the membrane-binding domain, and in the pattern of glycosylation. When expressed in Sf9 cells, the P. homomalla enzyme forms a 15R-prostaglandin endoperoxide together with 11R-hydroxyeicosatetraenoic acid and 15R-hydroxyeicosatetraenoic acid as by-products. The endoperoxide gives rise to 15R-prostaglandins and 12R-hydroxyheptadecatrienoic acid, identified by comparison to authentic standards. Evaluation of the structural differences of this 15R-COX isozyme should provide new insights into the substrate binding and stereospecificity of the dioxygenation reaction of arachidonic acid in the cyclooxygenase active site.

Identification of the zinc-dependent endothelial cell binding protein for high molecular weight kininogen and factor XII: identity with the receptor that binds to the globular "heads" of C1q (gC1q-R).

High molecular weight kininogen (HK) and factor XII are known to bind to human umbilical vein endothelial cells (HUVEC) in a zinc-dependent and saturable manner indicating that HUVEC express specific binding site(s) for those proteins. However, identification and immunochemical characterization of the putative receptor site(s) has not been previously accomplished. In this report, we have identified a cell surface glycoprotein that is a likely candidate for the HK binding site on HUVECs. When solubilized HUVEC membranes were subjected to an HK-affinity column in the presence or absence of 50 microM ZnCl2 and the bound membrane proteins eluted, a single major protein peak was obtained only in the presence of zinc. SDS/PAGE analysis and silver staining of the protein peak revealed this protein to be 33 kDa and partial sequence analysis matched the NH2 terminus of gC1q-R, a membrane glycoprotein that binds to the globular "heads" of C1q. Two other minor proteins of approximately 70 kDa and 45 kDa were also obtained. Upon analysis by Western blotting, the 33-kDa band was found to react with several monoclonal antibodies (mAbs) recognizing different epitopes on gC1q-R. Ligand and dot blot analyses revealed zinc-dependent binding of biotinylated HK as well as biotinylated factor XII to the isolated 33-kDa HUVEC molecule as well as recombinant gC1q-R. In addition, binding of 125I-HK to HUVEC cells was inhibited by selected monoclonal anti-gC1q-R antibodies. C1q, however, did not inhibit 125I-HK binding to HUVEC nor did those monoclonals known to inhibit C1q binding to gC1q-R. Taken together, the data suggest that HK (and factor XII) bind to HUVECs via a 33-kDa cell surface glycoprotein that appears to be identical to gC1q-R but interact with a site on gC1q-R distinct from that which binds C1q.

High speed imaging and algorithms for non invasive vibrations measurement

Comunicación presentada en EVACES 2011, 4th International Conference on Experimental Vibration Analysis for Civil Engineering Structures, Varenna (Lecco), Italy, October 3-5, 2011.

Temperature influence on the physical and mechanical properties of a porous rock: San Julian's calcarenite

This work discusses the results from tests which were performed in order to study the effect of high temperatures in the physical and mechanical properties of a calcarenite (San Julian's stone). Samples, previously heated at different temperatures (from 105 °C to 600 °C), were tested. Non-destructive tests (porosity and ultrasonic wave propagation) and destructive tests (uniaxial compressive strength and slake durability test) were performed over available samples. Furthermore, the tests were carried out under different conditions (i.e. air-cooled and water-cooled) in order to study the effect of the fire off method. The results show that uniaxial compressive strength and elastic parameters (i.e. elastic modulus and Poisson's ratio), decrease as the temperature increases for the tested range of temperatures. A reduction of the uniaxial compressive strength up to 35% and 50% is observed in air-cooled and water-cooled samples respectively when the samples are heated to 600 °C. Regarding the Young's modulus, a fall over 75% and 78% in air-cooled and water-cooled samples respectively is observed. Poisson's ratio also declines up to 44% and 68% with the temperature in air-cooled and water-cooled samples respectively. Slake durability index also exhibits a reduction with temperature. Other physical properties, closely related with the mechanical properties of the stone, are porosity, attenuation and propagation velocity of ultrasonic waves in the material. All exhibit considerable changes with temperature.

Regio and diastereoselective multicomponent 1,3-dipolar cycloadditions between prolinate hydrochlorides, aldehydes and dipolarophiles for the direct synthesis of pyrrolizidines

A general synthesis of highly substituted pyrrolizidines can be performed by a multicomponent 1,3-dipolar cycloaddition using proline ester hydrochlorides, aldehydes and dipolarophiles, at room temperature without catalysts or in the presence of AgOAc (5 mol %). In the case of (2S,4R)-4-hydroxyproline derivatives it is possible to obtain enantioenriched pyrrolizidines with high control of the regio- and diastereoselectivity affording the adducts 2,4-trans-2,5-trans according to an endo-approach and a S-dipole geometry of the in situ generated azomethine ylide. For proline esters a similar regioselectivity and endo-diastereoselectivity are observed when the dipole promotes an α-attack. However, when ethyl glyoxylate is used as aldehyde component the γ-attack of the S-ylide takes place preferentially giving rise the opposite regioselectivity for acrylic dipolarophiles, being crucial the role of silver acetate. In this case, the exo-adducts with a 2,3-cis-2,5-trans relative configuration are diastereoselectively obtained. In addition, computational studies have also been carried out to shed light on the origins of the diastereo- and regioselectivity observed for the described 1,3-dipolar cycloadditions.

Radon gas. Environmental concentration measures and remediation actions in exposed buildings

The main contribution to the radiological impact from natural radiation received by general population is due to the emission of radon (222Rn). The objective of this project is the study of radon gas as a radioactive element in our buildings (existing and future constructions) to avoid its influence in interior rooms. The proposed methodology reflects different aspects of natural radioactivity in buildings, their sources, their control criteria and regulatory framework; aspects related to the presence of radon in our constructions, entryways, measurement methodology for indoor environmental concentration are studied; other protection solutions and remediation measures in both existing buildings and new construction projects are analyzed. In conclusion, the paper presents previous evaluation tools, the analysis of existing concentration and the choice of the most appropriate mitigation / remediation measures depending on each case, through the establishment of different architectural proposals to plan actions against radon where necessary.

How buildings visualise client and architect: The problem that today’s user is typically not the client

An article presented at the last ICAT- conference stated at the end that buildings at all times tend to picture the people who had them erected. This paper aims to show the correctness of that statement. To this end, it will examine a number of typical residential buildings dating from the beginning of the seventeenth century up to today, investigate who had the buildings erected, and relate that to the performance of the buildings. This relation analysis will mainly use the scale but also the degree of diversity in function and appearance as factors. Furthermore, using economic data and data on the buildings to identify patterns, it will investigate how size of the property and relative size of the capital interest behind the building has developed. Since the authors live in Copenhagen and Copenhagen is very typical in its historical development, buildings and environments in and around the centre of Copenhagen are used as examples.

Micro-, Small- and Medium-Sized Enterprises with High-Growth Potential in the Southern Mediterranean: Identifying Obstacles and Policy Responses. CEPS Paperbacks. January 2014

The Arab Spring, which took root in Tunisia and Egypt in the beginning of 2011 and gradually spread to other countries in the southern Mediterranean, highlighted the importance of private-sector development, job creation, improved governance and a fairer distribution of economic opportunities. The developments led to domestic and international calls for the region’s governments to implement the needed reforms to enhance business and investment conditions, modernise their economies and support the development of enterprises. Central to these demands are calls to enhance the growth prospects of micro-, small- and medium-sized enterprises (MSMEs), which represent an overwhelming majority of the region’s economic activity.

Calculation of paleoproductivity during the Eocene-Oligocene transition of ODP Site 113-689

High-resolution records of carbon and oxygen isotopes and benthic foraminiferal accumulation rates for the Eocene-Oligocene section at Ocean Drilling Program Site 689 (Maud Rise, Weddell Sea; paleodepth about 1500 m) were used to infer variations in paleoproductivity in relation to changes in climate and ventilation of the deeper-water column. The benthic foraminiferal abundance and isotope records show short-term fluctuations at periodicities of 100 and 400 ka, implying orbitally driven climatic variations. Both records suggest that intermediate-depth water chemistry and primary productivity changed in response to climate. During the Eocene, productivity increased during cold periods and during cold-to-warm transitions, possibly as a result of increased upwelling of nutrient-rich waters. In the Oligocene, in contrast, productivity maxima occurred during intervals of low delta18O values (presumably warmer periods), when a proto-polar front moved to the south of the location of Site 689. This profound transition in climate-productivity patterns occurred around 37 Ma, coeval with rapid changes toward increasing variability of the oxygen and carbon isotope and benthic abundance records and toward larger-amplitude delta18O fluctuations. Therefore, we infer that, at this time, temperature fluctuations increased and a proto-polar front formed in conjunction with the first distinct pulsations in size of the Antarctic ice sheet. We speculate that this major change might have resulted from an initial opening of the Drake Passage at 37 Ma, at least for surface- and intermediate-water circulation.

Carbonate preservation reconstructed from ODP Sites 154-927 and 154-929 at Ceará Rise

Enhanced Atlantic overturning during the Pliocene was first proposed almost 10 yrs ago. Evidence for this Pliocene super conveyor scenario has been collected using a number of proxies (e.g., benthic d13C, Nd isotopic composition of manganese crusts). The present study contributes to the existing evidences by using carbonate dissolution and current vigour history of early Pliocene sediments from the Ceará Rise (ODP Sites 927 and 929). In order to reveal carbonate dissolution history, a number of commonly used and newly established proxies were applied, i.e., sand and carbonate contents, foraminifer fragmentation index, Bulloides Dissolution Index and carbonate silt grain-size distributions. Terrigenous silt grain-size distributions were used to unravel variations in relative current strength and sediment input to the two sites. Overall good carbonate preservation at the shallow Site 927 (3314 m water depth) shows that this level was bathed in North Atlantic Deep Water throughout the early Pliocene. The contrastingly poor carbonate preservation record of the deeper Site 929 (4358 m water depth, at present exposed to Antarctic Bottom Water) is frequently interrupted by phases of good carbonate preservation. These results indicate that the depth of the calcite lysocline was mainly tied to present level (ab. 4200 m water depth), and sometimes even dropped to water depths greater than 4360 m due to even more enhanced circulation. Surprisingly the expansion of NADW is not clearly reflected by an increase in current speed as shown by continuously fine terrigenous grain size.

Planktonic foraminiferal preservation in Plio/Pleistocene sediments from the western equatorial Atlantic and Caribbean

Records of mean sortable silt and planktonic foraminiferal preservation from the Ceará Rise (western equatorial Atlantic) and from the Caribbean are presented to analyze the Pliocene (3.5-2.2 Ma) to Pleistocene (1.6-0.3 Ma) evolution of near-bottom current strength and the carbonate corrosiveness of deep water. During the mid-Pleistocene climate transition (~1 Ma) a drastic decrease in glacial bottom current strength and an increase in carbonate corrosiveness is registered, demonstrating a substantial decrease in the glacial contribution of the Lower North Atlantic Deep Water (LNADW) to the Atlantic Ocean. Also, an increased sensitivity to eccentricity orbital forcing is registered after the MPT. By contrast, carbonate preservation increases considerably in the deep Caribbean in response to a strong and persistent stable contribution of Upper North Atlantic Deep Water (UNADW). We found evidence for the strongest and most stable circulation within the LNADW cell during the Northern Hemisphere cooling period between ~3.2 and 2.75 Ma. This is in agreement with the 'superconveyor model' which postulates that the highest NADW production took place prior to ~2.7 Ma. A considerable decrease in bottom current strength and planktonic foraminiferal preservation is observed synchronous with the first occurrence of large-scale continental ice sheets in the Northern Hemisphere. This documents the final termination of the 'superconveyor' at ca. 2.75 Ma. However, our data do not support a 'superconveyor' in the interval between 3.5 and 3.2 Ma when high-amplitude fluctuations in bottom current flow and preservation in planktonic foraminifera are observed. Because of the great sensitivity of NADW production to changes in surface water salinity, we assume that the high-amplitude fluctuations of LNADW circulation prior to ~3.2 Ma are linked to changes in the Atlantic salinity budget. After 2.75 Ma they are primarily controlled by ice-sheet forcing. In contrast to the stepwise deterioration of planktonic foraminiferal preservation in the western deep Atlantic, a trend toward better preservation from the Pliocene to Pleistocene is observed in the deep Caribbean. This indicates a long-term increase in the contribution of UNADW to the Atlantic Ocean.

Pliocene-Pleistocene stack of globally distributed benthic stable oxygen isotope records

We present a 5.3-Myr stack (the ''LR04'' stack) of benthic d18O records from 57 globally distributed sites aligned by an automated graphic correlation algorithm. This is the first benthic delta18O stack composed of more than three records to extend beyond 850 ka, and we use its improved signal quality to identify 24 new marine isotope stages in the early Pliocene. We also present a new LR04 age model for the Pliocene-Pleistocene derived from tuning the delta18O stack to a simple ice model based on 21 June insolation at 65 N. Stacked sedimentation rates provide additional age model constraints to prevent overtuning. Despite a conservative tuning strategy, the LR04 benthic stack exhibits significant coherency with insolation in the obliquity band throughout the entire 5.3 Myr and in the precession band for more than half of the record. The LR04 stack contains significantly more variance in benthic delta18O than previously published stacks of the late Pleistocene as the result of higher resolution records, a better alignment technique, and a greater percentage of records from the Atlantic. Finally, the relative phases of the stack's 41- and 23-kyr components suggest that the precession component of delta18O from 2.7-1.6 Ma is primarily a deep-water temperature signal and that the phase of d18O precession response changed suddenly at 1.6 Ma.

Oligocene to Miocene stable isotope record and planktonic foraminifer biostratigraphy of the Sierra Leone Rise

We have integrated Oligocene to lower upper Miocene planktonic foraminifer biostratigraphy with benthic foraminifer (Cibicidoides spp.) stable isotope records for two sites drilled on opposite sides of the Sierra Leone Rise in the eastern equatorial Atlantic Ocean. Deep Sea Drilling Project Site 366 (2853 m present water depth; 2200-2800 m paleodepth) recovered an Oligocene to upper Miocene record with a minor unconformity in the "middle" Oligocene and a condensed middle Miocene section. Ocean Drilling Program Site 667 (3529 m present depth; 3000-3500 m paleodepth) recovered an apparently continuous "middle" Oligocene to lower middle Miocene record and a similar condensed middle Miocene section. The Oligocene to lower Miocene sections were deposited at similar sedimentation rates (~11-16 m/m.y.). Stable isotope stratigraphy proved to be useful in establishing intra- and interbasinal correlations. In addition to the well-known earliest Oligocene and middle Miocene S180 increases, a distinct d18O increase occurred near the Oligocene/Miocene boundary. Carbon isotope variations provide similar potential for improving correlations; for example, a d13C increase occurred near the Oligocene/Miocene boundary in concert with increased d18O values. There was little d13C difference between the western Atlantic and eastern Atlantic basins during the late Oligocene and most of the middle Miocene; in contrast, eastern basin d13C values were slightly lower than those in the western basins during the earliest Oligocene (about 35-33 Ma) and early Miocene (about 22-18 Ma).

Maestrichtian calcareous nannofossils of MAud Rise, Weddell Sea

Recovery of an essentially complete upper Maestrichtian/lower Paleocene interval on Maud Rise at 65 °S latitude in the Weddell Sea during Ocean Drilling Program Leg 113 marks the first time that this interval has been cored at these high latitudes. The entire interval was missing at all Falkland Plateau sites drilled during DSDP Legs 36 and 71. Maestrichtian nannofossil assemblages in sediments from Sites 689 and 690, therefore, provide the basis for a needed revision of Maestrichtian coccolith zonation schemes for high southern latitudes. Three zones and two new subzones are described: the uppermost Maestrichtian Nephrolithus frequens Zone, which is subdivided into the Cribrosphaerella daniae Subzone and the underlying N. corystus Subzone, and the Biscutum magnum and B. coronum Zones. A complete calcareous nannofossil biostratigraphy based on the proposed scheme is given including a description of individual species abundance, preservation, and stratigraphic distribution. At this site, the southernmost carbonate site yet drilled by DSDP/ODP, it is evident that the Falkland Plateau Nannofossil Biogeographic Province can be extended to the margins of Antarctica. In addition, the biogeographic ranges of many calcareous nannofossils can likewise be extended. Last, we hypothesize that Nephrolithus frequens evolved from N. corystus prior to its dispersal to the lower latitudes where it is an important zonal marker. Three new taxa, Neocrepidolithus watkinsii n. sp., Nephrolithus frequens miniporus emend, n. comb, and Psyktosphaera firthii n. gen., n. sp. are described.