Lithogeochemical and isotope (Rb-Sr, Sm-Nd, Pb-Pb whole rock and Lu-Hf zircon) composition of samples from the Shackleton Range, East Antarctica

Three distinct, spatially separated crustal terranes have been recognised in the Shackleton Range, East Antarctica: the Southern, Eastern and Northern Terranes. Mafic gneisses from the Southern Terrane provide geochemical evidence for a within-plate, probably back-arc origin of their protoliths. A plume-distal ridge origin in an incipient ocean basin is the favoured interpretation for the emplacement site of these rocks at c. 1850 Ma, which, together with a few ocean island basalts, were subsequently incorporated into an accretionary continental arc/supra-subduction zone tectonic setting. Magmatic underplating resulted in partial melting of the lower crust, which caused high-temperature granulite-facies metamorphism in the Southern Terrane at c. 1710-1680 Ma. Mafic and felsic gneisses there are characterised by isotopically depleted, positive Nd and Hf initials and model ages between 2100 and 2000 Ma. They may be explained as juvenile additions to the crust towards the end of the Palaeoproterozoic. These juvenile rocks occur in a narrow, c. 150 km long E-W trending belt, inferred to trace a suture that is associated with a large Palaeoproterozoic accretionary orogenic system. The Southern Terrane contains many features that are similar to the Australo-Antarctic Mawson Continent and may be its furthermost extension into East Antarctica. The Eastern Terrane is characterised by metagranitoids that formed in a continental volcanic arc setting during a late Mesoproterozoic orogeny at c. 1060 Ma. Subsequently, the rocks experienced high-temperature metamorphism during Pan-African collisional tectonics at 600 Ma. Isotopically depleted zircon grains yielded Hf model ages of 1600-1400 Ma, which are identical to Nd model ages obtained from juvenile metagranitoids. Most likely, these rocks trace the suture related to the amalgamation of the Indo-Antarctic and West Gondwana continental blocks at ~600 Ma. The Eastern Terrane is interpreted as the southernmost extension of the Pan-African Mozambique/Maud Belt in East Antarctica and, based on Hf isotope data, may also represent a link to the Ellsworth-Whitmore Mountains block in West Antarctica and the Namaqua-Natal Province of southern Africa. Geochemical evidence indicates that the majority of the protoliths of the mafic gneisses in the Northern Terrane formed as oceanic island basalts in a within-plate setting. Subsequently the rocks were incorporated into a subduction zone environment and, finally, accreted to a continental margin during Pan-African collisional tectonics. Felsic gneisses there provide evidence for a within-plate and volcanic arc/collisional origin. Emplacement of granitoids occurred at c. 530 Ma and high-temperature, high-pressure metamorphism took place at 510-500 Ma. Enriched Hf and Nd initials and Palaeoproterozoic model ages for most samples indicate that no juvenile material was added to the crust of the Northern Terrane during the Pan-African Orogeny but recycling of older crust or mixing of crustal components of different age must have occurred. Isotopically depleted mafic gneisses, which are spatially associated with eclogite-facies pyroxenites, yielded late Mesoproterozoic Nd model ages. These rocks occur in a narrow, at least 100 km long, E-W trending belt that separates alkaline ocean island metabasalts and within-plate metagranitoids from volcanic arc metabasalts and volcanic arc/syn-collisional metagranitoids in the Northern Terrane. This belt is interpreted to trace the late Neoproterozoic/early Cambrian Pan-African collisional suture between the Australo-Antarctic and the combined Indo-Antarctic/West Gondwana continental blocks that formed during the final amalgamation of Gondwana.

Strontium- and lead-isotope composition of some basalts at DSDP Hole 69-504B

Seventeen whole-rock samples, generally taken at 25- to 50-meter intervals from 5 to 560 meters sub-basement in Deep Sea Drilling Project Hole 504B, were analyzed for 87Sr/86Sr ratios, and rubidium and strontium concentrations. Ten of these samples also were analyzed for Pb-isotope composition. Strontium-isotope ratios for eight samples from the upper 260 meters of the hole range from 0.70287 to 0.70377, with a mean of 0.70320. In the interval 330 to 560 meters, five samples have a restricted range of 0.70259 to 0.70279, with a mean of 0.70266, almost identical to the average value of fresh mid-ocean-ridge basalts. In the interval 260 to 330 meters, approximately intermediate strontium- isotope ratios are found. The higher 87Sr/86Sr ratios in the upper part of the hole can be interpreted in terms of strontium-isotope alteration during basalt-sea-water interaction. Relative to average fresh mid-ocean ridge basalts, the upper 260 meters of basalts are enriched by an average of about 9% in sea-water strontium 87Sr/86Sr = 0.7091). This Sr presumably is located in the smectites, which, as the main secondary minerals throughout the hole, replace olivine and matrix glass and locally fill vesicles (analyzed samples contained no veins). The strontium-isotope data strongly suggest that the integrated flux of sea water through the upper part of the Hole 504B crust has been greater than through the lower part. This is also suggested by (1) the common occurrence of Feoxide- hydroxide minerals as alteration products above 270 meters, but their near absence below 320 meters, (2) the presence of vein calcite above 320 meters, but its near absence below this level, and (3) the occurrence of vein pyrite only below a depth of 270 meters. Sea-water circulation in the lower basalts may have been partly restricted by the greater number of relatively impermeable massive lava flows below 230 meters sub-basement. Although sufficient sea water was present within the lower part of the hole to produce smectitic alteration products, the overall water /rock ratio was low enough to prevent significant modification of strontium-isotope ratios. Lead-isotope ratios of Hole 504B basalts form approximately linear arrays in plots of 208Pb/204Pb and 207Pb/204Pb versus 206Pb/204Pb. The arrays are similar to those reported for basalts from other mid-ocean ridges. There is no trend in Hole 504B lead-isotope ratios with vertical position in the basement. The arrays indicate that the lead-isotope composition of the upper mantle from which the Hole 504B basaltic melts were derived was inhomogeneous.

Major element oxides and 40Ar/39Ar plateau and isochron ages of ODP Hole 165-1001A

40Ar-39Ar incremental heating experiments and electron microprobe analyses were performed on basaltic rocks recovered from Site 1001 during Ocean Drilling Program Leg 165. The lower Nicaraguan Rise, on which Site 1001 lies, appears to be part of a larger Caribbean oceanic plateau that makes up the core of the Caribbean plate. Our results indicate an eruption age of 81 ± 1 Ma. A single flow-rim glass is tholeiitic and almost identical to the shipboard X-ray fluorescence analyses of the whole rock. The slightly porphyritic basalts have at least two populations of plagioclase, groundmass, and glomerocrystic plagioclase laths that appear to be in equilibrium with the surrounding melt and corroded tabular phenocrysts that have a higher An content (An84-86).

Shallow water foraminifera of ODP Hole 133-821A

Reworked shallow-water foraminifers that settled on the upper slope of the central Great Barrier Reef at Site 821 (water depth, 212.6 m) were used as indicators of the paleoclimatic and paleoenvironmental conditions that have controlled the Pleistocene evolution of the adjacent platform. Throughout the 400-m-thick sequence drilled, the nature, composition, and distribution of the shallow-water foraminiferal assemblages studied indicate that (1) all the species recorded are at present living in diverse tropical, reef-related areas of the Indo-Pacific and Atlantic provinces; (2) the composition of the microfaunal taphocoenoses is almost identical between the different stratigraphic intervals studied and the modern Great Barrier Reef environments; (3) inner-neritic, tropical environments have continued to develop since the middle Pleistocene; (4) high- to moderate-energy platform edges occurred repeatedly throughout Pleistocene time. These factors may suggest that, since the beginning of the Pleistocene, several reef-like tracts have grown successively on the central area of the northeastern Australian shelf edge. These tracts probably had a sufficiently evolved morphological zonation to act as shelters for foraminiferal biocoenoses of high species diversity.

Description and chemical composition of manganese nodules during R/V "Tyro" cruise APNAP 1 to the Madeira Abyssal Plain in 1986

The porewater and sediment composition of two boxcores and of a small gravity core, taken on a manganese-nodule-covered hill and in the Madeira Abyssal Plain proper respectively, are compared. The pore-water study of the two boxcores indicates that oxic conditions prevail in both cores. In addition, it indicates that no detectable fluxes of Mn or Fe occur from the porewater to the ocean bottom water. Variations in the geochemical composition of the sediments can be explained by fluctuations in the amount of carbonate, which acts as a diluting agent. A clear carbonate minimum is observed at 20-22 cm depth in the two cores. This minimum is likely to be associated with the last glacial period (10-20 kyr B.P.). This association is supported by the sediment accumulation rate of 15 mm/kyr as found by extrapolation from the rate for pelagic sediments in the Madeira Abyssal Plain. The bulk composition of the manganese nodules recovered from the submarine hill is chemically almost identical to the average composition of Atlantic nodules. The trace metal and Rare Earth Elements composition indicate a hydrogenous origin for the manganese nodules of this study. On the basis of the chemical composition, and that of nodules relative to that of the adjacent sediments, an average nodule accretian rate of 2.8-3.3 mm/myr has been calculated. Although the analyses of the entire ferromanganese nodules that have been studied seem to indicate a homogenous composition, internal structures of the nodules reveal great inhomogeneity, both visually and chemically. These fluctuations may be related to variations in the fluxes of Mn and Fe, which in turn could be climate-related.

Benthic foraminifera stable isotope data from the South Pacific sediment cores PS75/059-2 and PS75/056-1

Investigating the inter-basin deep water exchange between the Pacific and Atlantic Oceans over glacial-interglacial climate cycles is important for understanding circum-Antarctic Southern Ocean circulation changes and their impact on the global Meridional Overturning Circulation. We use benthic foraminiferal d13C records from the southern East Pacific Rise to characterize the d13C composition of Circumpolar Deep Water in the South Pacific, prior to its transit through the Drake Passage into the South Atlantic. A comparison with published South Atlantic deep water records from the northern Cape Basin suggests a continuous water mass exchange throughout the past 500 ka. Almost identical glacial-interglacial d13C variations imply a common deep water evolution in both basins suggesting persistent Circumpolar Deep Water exchange and homogenization. By contrast, deeper abyssal waters occupying the more southern Cape Basin and the southernmost South Atlantic have lower d13C values during most, but not all, stadial periods. We conclude that these values represent the influence of a more southern water mass, perhaps AABW. During many interglacials and some glacial periods, the gradient between Circumpolar Deep Water and the deeper southern Cape Basin bottom water disappears suggesting either no presence of AABW or indistinguishable d13C values of both water masses.

Oxygen isotopes and hydrocarbon composition of gas hydrate and hydrate water from ODP Leg 165 sites

Ocean Drilling Program (ODP) Leg 164 recovered a number of large solid gas hydrate from Sites 994, 996, and 997 on the Blake Ridge. Sites 994 and 997 samples, either nodular or thick massive pieces, were subjected to laboratory analysis and measurements to determine the structure, molecular and isotopic composition, thermal conductivity, and equilibrium dissociation conditions. X-ray computed tomography (CT) imagery, X-ray diffraction, nuclear magnetic resonance (NMR), and Raman spectroscopy have revealed that the gas hydrates recovered from the Blake Ridge are nearly 100% methane gas hydrate of Structure I, cubic with a lattice constant of a = 11.95 ± 0.05 angström, and a molar ratio of water to gas (hydration number) of 6.2. The d18O of water is 2.67 per mil to 3.51 per mil SMOW, which is 3.5-4.0 heavier than the ambient interstitial waters. The d13C and dD of methane are -66 per mil to -70 per mil and -201 per mil to -206 per mil, respectively, suggesting that the methane was generated through bacterial CO2 reduction. Thermal conductivity values of the Blake Ridge hydrates range from 0.3 to 0.5 W/(m K). Equilibrium dissociation experiments indicate that the three-phase equilibrium for the specimen is 3.27 MPa at 274.7 K. This is almost identical to that of synthetic pure methane hydrate in freshwater.

Seawater carbonate chemistry and community calcification during Miyako Island (Japan) coral reef studies, 1994

Coral reefs are characterized by enormous carbonate production of the organisms. It is known that rapid calcification is linked to photosynthesis under control of the carbonate equilibrium in seawater. We have established a model simulating the coexisting states of photosynthesis and calcification in order to examine the effects of photosynthesis and calcification on the carbonate system in seawater. Supposing that the rates of photosynthesis and calcification are proportional to concentrations of their inorganic carbon source, the model calculations indicate that three kinds of unique interactions of the organic and inorganic carbon productions are expected. These are photosynthetic enhancement of calcification, calcification which benefits photosynthesis and carbonate dissolution induced by respiration. The first effect appears when the photosynthetic rate is more than approximately 1.2 larger than that of calcification. This effect is caused by the increase of CO3 content and carbonate saturation degree in seawater. If photosynthesis use molecular carbon dioxide, the second effect occurs when the calcification rate is more than approximately 1.6 times larger than that of photosynthesis. Time series model experiments indicate that photosynthesis and calcification potentially enhance each other and that organic and inorganic carbon is produced more efficiently in the coexisting system than in the isolated reactions. These coexisting effects on production enhancement of photosynthesis and calcification are expected to appear not only in the internal pool of organisms but also in a reef environment which is isolated from the outer ocean during low tide. According to the measurements on the fringing type Shiraho Reef in the Ryukyu Islands, the diurnal change of water properties (pH, total alkalinity, total carbon dioxide and carbonate saturation degree) were conspicuous. This environment offers an appropriate condition for the appearance of these coexisting effects. The photosynthetic enhancement of calcification and the respiratory inducement of decalcification were observed during day-time and night-time slack-water periods, respectively. These coexisting effects, especially the photosynthetic enhancement of calcification, appear to play important roles for fluorishing coral reef communities.

Toward an inclusive and a little bit ethical world trading system : listening to the voices of the people in LDCs

Many specialists in international trade have started saying that the era of a mega FTA is approaching. If the three poles of the global economy, namely East Asia, EU and the United States, form mega FTAs, most of the volume of global trade will be covered. That may be fine, but there will be many countries left out of the mega FTA, most of which will be the least developed countries (LDCs). Since the inception of the Doha Development Agenda (DDA) negotiations in 2001, the WTO and its member countries have tried to include LDCs in the world trading system through various means, including DFQF and AfT. Although these means have some positive impact on the economic development of LDCs, most of the LDCs will never feel comfortable with the current world trading system. To overcome the stalemate in the DDA and to create an inclusive world trading system, we need more commitment from both LDCs and non-LDCs. To surmount the prolonged stalemate in the DDA, we should understand how ordinary people in LDCs feel and think about the current world trading system. Those voices have seldom been listened to, even by the decision makers of their own countries. So as to understand the situation of the people in LDCs, IDE-JETRO carried out several research projects using macro, meso and micro approaches. For the micro level, we collected and analyzed statements from ordinary people concerning their opinions about the world trading system. The interviewees are ordinary people such as street vendors, farmers and factory workers. We asked about where they buy and sell daily necessities, their perception of imported goods, export promotion and free trade at large, etc. These ‘voices of the people’ surveys were conducted in Madagascar and Cambodia during 2013. Based on this research, and especially the findings from the ‘voices of the people’ surveys, we propose a ‘DDA-MDGs hybrid’ strategy to conclude DDA negotiations and develop a more inclusive and a little bit more ethical world trading system. Our proposal may be summarized in the following three points. (1) Aid for Trade (AfT) ver. 2 Currently AfT is mainly focused on coordinating several aid projects related to LDCs’ capacity building. However, this is inadequate; for the proposed ‘DDA-MDGs hybrid’, a super AfT is needed. The WTO, other development agencies and LDC governments will not only coordinate but also plan together aid projects for trade capacity building. AfT ver. 2 includes infrastructure projects either gran aid, ODA loans and private investment. This is in accordance with the post-MDGs argument which emphasizes the role of the private sector. (2) Ethical Attitude Reciprocity is a principle of multilateral agreement, and it has been a core promise since GATT. However, for designing an inclusive system, special and differential treatment (S&D) is still needed for disadvantaged members. To compromise full reciprocity and less than full reciprocity, an ethical attitude on the part of every member is needed in which every member refrains from insisting on the full rights and demands of its own country. As used herein, the term ‘ethical’ implies more consideration for LDCs, and it is almost identical to S&D but with a more positive attitude from developed countries (super S&D). (3) Collect Voices of the People In order to grasp the real situation of the people, the voices of the people on free trade will continue to be collected in other LDCs, and the findings and leanings will be fed back to the WTO negotiation space.

Multi Domain-Specific Modeling of the Security Concerns of Service-Oriented Architectures

Service-Oriented Architectures (SOA), and Web Services (WS), the technology generally used to implement them, achieve the integration of heterogeneous technologies, providing interoperability, and yielding the reutilization of pre-existent systems. Model-driven development methodologies provide inherent benefits such as increased productivity, greater reuse, and better maintainability, to name a few. Efforts on achieving model-driven development of SOAs already exist, but there is currently no standard solution that addresses non-functional aspects of these services as well. This paper presents an approach to integrate these non-functional aspects in the development of web services, with an emphasis on security.

DNS of a hot-wire anemometer in a turbulent channel flow

A body with a shape similar to a hot wire with its sheath, but no prongs, has been placed close to the wall of a turbulent channel at Re_tau = 600. The results of the channel flow, without the wire, agree with previous published ones, despite the modest resolution and domain size. A simplified, two-dimensional version of the wire at the same Reynolds number has been studied to compare the dynamic response of cold and hot wires, where a slightly bigger perturbation is seen in the hot case, but an almost identical dynamic response. The cold wire seems to be able to measure instantaneous velocity with total drag after proper calibration. Being a DNS, the complete description of the flow field around the wire is obtained.

Jacques Tati : vivienda experimental y espacio de trabajo a mediados del siglo XX

La obra fílmica del director francés Jacques Tati podría considerarse como el perfecto reflejo del paradigma edificatorio de mediados del siglo XX en plena posguerra europea, una época ávida de transformaciones de las que el cine supo hacerse eco. Particularmente, el cine de Tati refleja las preocupaciones del ciudadano europeo de posguerra sobre las consecuencias de las masivas construcciones erigidas en sus devastados núcleos urbanos y la puesta en práctica de la ciudad funcional propuesta por la Carta de Atenas (1931). Pero, además, el análisis del cine de Jacques Tati permite un acercamiento a la modernidad desde diversos puntos de vista como la movilidad, el diseño urbano, las nuevas construcciones, los espacios de trabajo en los nuevos distritos terciarios, la vivienda -tradicional, moderna y experimental- o el diseño mobiliario en la posguerra. A través de su alter ego -Monsieur Hulot- Tati interacciona curioso con las nuevas construcciones geométricas de paños neutros y cuestiona su ruptura con la tradición edificatoria francesa, enfatizando la oposición entre el pasado nostálgico y la modernidad de las décadas de los 50 y 60, salpicadas por el consumismo feroz del recién estrenado estado de bienestar. La confrontación funcional, volumétrica, estética e incluso cromática entre ambos mundos construidos –el tradicional y el moderno- invita al espectador a un ejercicio de reflexión y crítica sobre la arquitectura moderna de este período en Europa. En particular, la mirada cinematográfica de Tati se centra en dos conceptos fundamentales. Por una parte, su atención se dirige a la famosa casa mecanicista Le Corbuseriana materializada en la ultra-moderna casa Arpel (Mon Oncle, 1958) y proyectada en la misma época en la que se desarrollaban importantes prototipos de vivienda experimental como la Casa de Futuro de Alison y Peter Smithson o las viviendas de Jean Prouvé. Debe ponerse de manifiesto que la crítica de Jacques Tati no se centraba en la arquitectura moderna en sí misma sino en el empleo erróneo que los usuarios pudieran hacer de ella. Por otro lado, Tati centra su atención en el prisma miesiano a través de los bloques de oficinas que conforman la ciudad de Tativille en Playtime (1967). Se trataba de una gran ciudad moderna construida explícitamente para el rodaje de la película y basada en los casi idénticos tejidos urbanos residenciales y terciarios ya en funcionamiento en las principales capitales europeas y norteamericanas en aquellos años. Tativille funcionaría como una ciudad autónoma disponiendo de diversas instalaciones y con el objetivo de integrarse y consolidarse en la trama urbana parisina. Lamentablemente, su destino al final del rodaje fue bien distinto. En definitiva, el análisis de la producción fílmica de Jacques Tati permite un acercamiento a la arquitectura y al urbanismo modernos de posguerra y al contexto socio-económico que favoreció su crecimiento y expansión. Por ello, su obra constituye una herramienta visual muy útil que aún hoy es consultada y mostrada por su claridad y humor y que invita a los ciudadanos –telespectadores- a participar en un ejercicio crítico arquitectónico hasta entonces reservado a los arquitectos. ABSTRACT The film work of French director Jacques Tati could be considered as the perfect reflection of the mid-20th century European post-war building paradigm, a period of time plenty of transformations perfectly echoed by cinema. In particular, Tati’s film work reflects the European post-war citizen’s concerns about the consequences of massive constructions built in their desvastated urban centres, as well as the development of functional cities proposed by the Athens Charter (1931). But, on top of that, an analysis of Jacques Tati’s cinematography allows for an approach to modernity from different perspectives, such as mobility, urban design, new buildings, working spaces in the new tertiary districts, housing -traditional, modern, and experimental-, or furniture design during the post-war period. Embodied by his alter-ego –Monsieur Hulot,- Tati curiously interacts with the new geometric constructions of neutral facades and questions the break with the French building tradition, highlighting the opposition between the nostalgic past and modernity of the 50s and 60s, affected by the fierce consumerism of the new welfare state. The functional, volumetric, aesthetic and even chromatic confrontation between both built worlds –traditional vs modern- invites the viewer to an exercise of meditation and criticism on the European modern architecture of that period. Tati’s film look is particularly focused on two basic concepts: on the one hand, his attention addresses Le Corbusier’s famous mechanistic house which is materialized in the ultra-modern Arpel house (Mon Oncle, 1958) and designed, in turn, when the development of other important experimental dwelling prototypes like Alison and Peter Smithson’s House of the Future or Jean Prouvé´s houses was taking place. It must be highlighted that Jacques Tati’s criticism was not addressed to modern architecture itself but to the wrong use that citizens could make of it. On the other hand, Tati focuses on the Miesian prism through the office buildings that shape the city of Tativille in Playtime (1967). It was a big, modern city built specifically for the film shooting, and based on the almost identical residential and tertiary urban fabrics already active in the main European and American capitals those years. Tativille would work as an autonomous city, having several facilities at its disposal and with the goal of getting integrated and consolidated into the Parisian urban weave. However, its final use was, unfortunately, quite different. In conclusion, an analysis of Jacques Tati’s film production allows for an approach to modern post-war architecture and urbanism, as well as to the socio-economic context that favoured its growth and expansion. As a result of this, Jacques Tati’s film production constitutes a suitable visual tool which, even nowadays, is consulted and shown due to its clarity and humour, and at the same time invites citizens –viewers- to participate in an architectural criticism exercise that, so far, had been reserved to architects.

Direct interaction of flagellin termini essential for polymorphic ability of flagellar filament

We report the structures of flagellar filaments reconstituted from various flagellins with small terminal truncations. Flagellins from Salmonella typhimurium strains SJW1103 (wild type), SJW1660, and SJW1655 were used, which form a left-handed supercoil, the L- and R-type straight forms, respectively. Structure analyses were done by electron cryomicroscopy and helical image reconstruction with a help of x-ray fiber diffraction for determining precise helical symmetries. Truncation of either terminal region, irrespective of the original flagellin species, results in a straight filament having a helical symmetry distinct either from the L- or R-type. This filament structure is named Lt-type. Although the local subunit packing is similar in all three types, a close comparison shows that the Lt-type packing is almost identical to the R-type but distinct from the L-type, which demonstrates the strong two-state preference of the subunit interactions. The structure clearly suggests that both termini are located in the inner tube of the concentric double-tubular structure of the filament core, and their proper interaction is responsible for the correct folding of fairly large terminal regions that form the inner tube. The double tubular structure appears to be essential for the polymorphic ability of flagellar filaments, which is required for the swimming–tumbling of bacterial taxis.

The nature of the excited state of the reaction center of photosystem II of green plants: A high-resolution fluorescence spectroscopy study

We studied the electronically excited state of the isolated reaction center of photosystem II with high-resolution fluorescence spectroscopy at 5 K and compared the obtained spectral features with those obtained earlier for the primary electron donor. The results show that there is a striking resemblance between the emitting and charge-separating states in the photosystem II reaction center, such as a very similar shape of the phonon wing with characteristic features at 19 and 80 cm−1, almost identical frequencies of a number of vibrational modes, a very similar double-Gaussian shape of the inhomogeneous distribution function, and relatively strong electron-phonon coupling for both states. We suggest that the emission at 5 K originates either from an exciton state delocalized over the inactive branch of the photosystem or from a fraction of the primary electron donor that is long-lived at 5 K. The latter possibility can be explained by a distribution of the free energy difference of the primary charge separation reaction around zero. Both possibilities are in line with the idea that the state that drives primary charge separation in the reaction center of photosystem II is a collective state, with contributions from all chlorophyll molecules in the central part of the complex.

Measurement of 8-hydroxy-2′-deoxyguanosine in DNA by high-performance liquid chromatography-mass spectrometry: comparison with measurement by gas chromatography-mass spectrometry

Measurement of 8-hydroxy-2′-deoxyguanosine (8-OH-dGuo) in DNA by high-performance liquid chromatography/mass spectrometry (LC/MS) was studied. A methodology was developed for separation by LC of 8-OH-dGuo from intact and modified nucleosides in DNA hydrolyzed by a combination of four enzymes: DNase I, phosphodiesterases I and II and alkaline phosphatase. The atmospheric pressure ionization-electrospray process was used for mass spectral measurements. A stable isotope-labeled analog of 8-OH-dGuo was used as an internal standard for quantification by isotope-dilution MS (IDMS). Results showed that LC/IDMS with selected ion-monitoring (SIM) is well suited for identification and quantification of 8-OH-dGuo in DNA at background levels and in damaged DNA. The sensitivity level of LC/IDMS-SIM was found to be comparable to that reported previously using LC-tandem MS (LC/MS/MS). It was found that approximately five lesions per 106 DNA bases can be detected using amounts of DNA as low as 2 µg. The results also suggest that this lesion may be quantified in DNA at levels of one lesion per 106 DNA bases, or even lower, when more DNA is used. Up to 50 µg of DNA per injection were used without adversely affecting the measurements. Gas chromatography/isotope-dilution MS with selected-ion monitoring (GC/IDMS-SIM) was also used to measure this compound in DNA following its removal from DNA by acidic hydrolysis or by hydrolysis with Escherichia coli Fpg protein. The background levels obtained by LC/IDMS-SIM and GC/IDMS-SIM were almost identical. Calf thymus DNA and DNA isolated from cultured HeLa cells were used for this purpose. This indicates that these two techniques can provide similar results in terms of the measurement of 8-OH-dGuo in DNA. In addition, DNA in buffered aqueous solution was damaged by ionizing radiation at different radiation doses and analyzed by LC/IDMS-SIM and GC/IDMS-SIM. Again, similar results were obtained by the two techniques. The sensitivity of GC/MS-SIM for 7,8-dihydro-8-oxoguanine was also examined and found to be much greater than that of LC/MS-SIM and the reported sensitivity of LC/MS/MS for 8-OH-dGuo. Taken together, the results unequivocally show that LC/IDMS-SIM is well suited for sensitive and accurate measurement of 8-OH-dGuo in DNA and that both LC/IDMS-SIM and GC/IDMS-SIM can provide similar results.