ALBERT

Description: The Aegean water masses and circulation structure are studied via two large-scale surveys performed during the late winters of 1988 and 1990 by the R/V Yakov Gakkel of the former Soviet Union. The analysis of these data sheds light on the mechanisms of water mass formation in the Aegean Sea that triggered the outflow of Cretan Deep Water (CDW) from the Cretan Sea into the abyssal basins of the eastern Mediterranean Sea (the so-called Eastern Mediterranean Transient). It is found that the central Aegean Basin is the site of the formation of Aegean Intermediate Water, which slides southward and, depending on their density, renews either the intermediate or the deep water of the Cretan Sea. During the winter of 1988, the Cretan Sea waters were renewed mainly at intermediate levels, while during the winter of 1990 it was mainly the volume of CDW that increased. This Aegean water mass redistribution and formation process in 1990 differed from that in 1988 in two major aspects: (i) during the winter of 1990 the position of the front between the Black Sea Water and the Levantine Surface Water was displaced farther north than during the winter of 1988 and (ii) heavier waters were formed in 1990 as a result of enhanced lateral advection of salty Levantine Surface Water that enriched the intermediate waters with salt. In 1990 the 29.2 isopycnal rose to the surface of the central basin and a large volume of CDW filled the Cretan Basin. It is found that, already in 1988, the 29.2 isopycnal surface, which we assume is the lowest density of the CDW, was shallower than the Kassos Strait sill and thus CDW egressed into the Eastern Mediterranean.

Description: Published

Description: 1841-1859

Description: JCR Journal

Description: reserved

Description: Five non-eddy-resolving oceanic general circulation models driven by atmospheric fluxes derived from the NCEP reanalysis are used to investigate the link between the Gulf Stream (GS) variability, the atmospheric circulation, and the Atlantic meridional overturning circulation (AMOC). Despite the limited model resolution, the temperature at the 200-m depth along the mean GS axis behaves similarly in most models to that observed, and it is also well correlated with the North Atlantic Oscillation (NAO), indicating that a northward (southward) GS shift lags a positive (negative) NAO phase by 0–2 yr. The northward shift is accompanied by an increase in the GS transport, and conversely the southward shift with a decrease in the GS transport. Two dominant time scales appear in the response of the GS transport to the NAO forcing: a fast time scale (less than 1 month) for the barotropic component, and a slower one (about 2 yr) for the baroclinic component. In addition, the two components are weakly coupled. The GS response seems broadly consistent with a linear adjustment to the changes in the wind stress curl, and evidence for baroclinic Rossby wave propagation is found in the southern part of the subtropical gyre. However, the GS shifts are also affected by basin-scale changes in the oceanic conditions, and they are well correlated in most models with the changes in the AMOC. A larger AMOC is found when the GS is stronger and displaced northward, and a higher correlation is found when the observed changes of the GS position are used in the comparison. The relation between the GS and the AMOC could be explained by the inherent coupling between the thermohaline and the wind-driven circulation, or by the NAO variability driving them on similar time scales in the models.

Description: This research was supported by the PREDICATE project of the European Community, and for M. Bentsen by the Research Council of Norway through RegClim, NOClim, and the Programme of Supercomputing.

Description: Published

Description: 2119–2135

Description: 3.7. Dinamica del clima e dell'oceano

Description: JCR Journal

Description: reserved

Description: A land surface model (LSM) has been included in the ECMWF Hamburg version 4 (ECHAM4) atmospheric general circulation model (AGCM). The LSM is an early version of the Organizing Carbon and Hydrology in Dynamic Ecosystems (ORCHIDEE) and it replaces the simple land surface scheme previously included in ECHAM4. The purpose of this paper is to document how a more exhaustive consideration of the land surface–vegetation processes affects the simulated boreal summer surface climate. To investigate the impacts on the simulated climate, different sets of Atmospheric Model Intercomparison Project (AMIP)-type simulations have been performed with ECHAM4 alone and with the AGCM coupled with ORCHIDEE. Furthermore, to assess the effects of the increase in horizontal resolution the coupling of ECHAM4 with the LSM has been implemented at different horizontal resolutions. The analysis reveals that the LSM has large effects on the simulated boreal summer surface climate of the atmospheric model. Considerable impacts are found in the surface energy balance due to changes in the surface latent heat fluxes over tropical and midlatitude areas covered with vegetation. Rainfall and atmospheric circulation are substantially affected by these changes. In particular, increased precipitation is found over evergreen and summergreen vegetated areas. Because of the socioeconomical relevance, particular attention has been devoted to the Indian summer monsoon (ISM) region. The results of this study indicate that precipitation over the Indian subcontinent is better simulated with the coupled ECHAM4–ORCHIDEE model compared to the atmospheric model alone.

Description: Published

Description: 255–278

Description: 3.7. Dinamica del clima e dell'oceano

Description: JCR Journal

Description: partially_open

Description: In this paper results from the application of an ocean data assimilation (ODA) system, combining a multivariate reduced-order optimal interpolator (OI) scheme with a global ocean general circulation model (OGCM), are described. The present ODA system, designed to assimilate in situ temperature and salinity observations, has been used to produce ocean reanalyses for the 1962–2001 period. The impact of assimilating observed hydrographic data on the ocean mean state and temporal variability is evaluated. A special focus of this work is on the ODA system skill in reproducing a realistic ocean salinity state. Results from a hierarchy of different salinity reanalyses, using varying combinations of assimilated data and background error covariance structures, are described. The impact of the space and time resolution of the background error covariance parameterization on salinity is addressed.

Description: This work has been funded by the ENACT Project (Contract EVK2-CT2001-00117) for A. Bellucci and P. Di Pietro, and partially by the ENSEMBLES Project (Contract GOCE-CT-2003-505539) for A. Bellucci.

Description: Published

Description: 3785-3807

Description: 3.7. Dinamica del clima e dell'oceano

Description: JCR Journal

Description: reserved

Description: The effect of horizontal resolution on tropical variability is investigated within the modified SINTEX model, SINTEX-F, developed jointly at INGV, IPSL and at the Frontier Research System. The horizontal resolutions T30 and T106 are investigated in terms of the coupling characteristics, frequency and variability of the tropical ocean-atmosphere interactions. It appears that the T106 resolution is generally beneficial even if it does not eliminate all the major systematic errors of the coupled model. There is an excessive shift west of the cold tongue and ENSO variability, and high resolution has also a somewhat negative impact to the variability in the East Indian Ocean. A dominant two-year peak for the NINO3 variabilty in the T30 model is moderated in the T106 as it shifts to longer time scale. At high resolution new processes come into play, as the coupling of tropical instability waves, the resolution of coastal flows at the Pacific Mexican coasts and improved coastal forcing along the coast of South America. The delayed oscillator seems the main mechanism that generates the interannual variability in both models, but the models realize it in different ways. In the T30 model it is confined close to the equator, involving relatively fast equatorial and near-equatorial modes, in the high resolution, it involves a wider latitudinal region and slower waves. It is speculated that the extent of the region that is involved in the interannual variability may be linked to the time scale of the variability itself.

Description: This research was partially supported by the Italy–USA Cooperation Program of the Italian Ministry of Environment and by the EU projects ENSEMBLES and DYNAMITE.

Description: Published

Description: 730-750

Description: 3.7. Dinamica del clima e dell'oceano

Description: JCR Journal

Description: reserved

Description: The Indian Summer Monsoon (ISM) is one of the main components of the Asian summer monsoon. It is well known that one of the starting mechanisms of a summer monsoon is the thermal contrast between land and ocean and that sea surface temperature (SST) and moisture are crucial factors for its evolution and intensity. The Indian Ocean, therefore, may play a very important role in the generation and evolution of the ISM itself. A coupled general circulation model, implemented with a high resolution atmospheric component, appears to be able to simulate the Indian summer monsoon in a realistic way. In particular, the features of the simulated ISM variability are similar to the observations. In this study, the relationships between ISM and Tropical Indian Ocean (TIO) SST anomalies are investigated, as well as the ability of the coupled model to capture those connections. The recent discovery of the Indian Ocean Dipole Mode (IODM) may suggest new perspectives in the relationship between ISM and TIO SST. A new statistical technique, the Coupled Manifold, is used to investigate the TIO SST variability and its relation with the Tropical Pacific Ocean (TPO). The analysis shows that the SST variability in the TIO contains a significant portion that is independent from the TPO variability. The same technique is used to estimate the amount of Indian rainfall variability that can be explained by the Tropical Indian Ocean SST. Indian Ocean SST anomalies are separated in a part remotely forced from the Tropical Pacific Ocean variability and a part independent from that. The relationships between the two SSTA components and the Indian monsoon variability are then investigated in detail.

Description: Published

Description: 3083-3105

Description: 3.7. Dinamica del clima e dell'oceano

Description: JCR Journal

Description: reserved

Description: An assessment of the present European operational marine monitoring and forecasting systems shows how observations, atmospheric forcing fields and ocean models combine to make useful oceanographic products possible.

Description: Published

Description: 1081-1090

Description: open

Description: Ensemble experiments are performed with five coupled atmosphere–ocean models to investigate the potential for initial-value climate forecasts on interannual to decadal time scales. Experiments are started from similar model-generated initial states, and common diagnostics of predictability are used. We find that variations in the ocean meridional overturning circulation (MOC) are potentially predictable on interannual to decadal time scales, a more consistent picture of the surface temperature impact of decadal variations in the MOC is now apparent, and variations of surface air temperatures in the North Atlantic Ocean are also potentially predictable on interannual to decadal time scales, albeit with potential skill levels that are less than those seen for MOC variations. This intercomparison represents a step forward in assessing the robustness of model estimates of potential skill and is a prerequisite for the development of any operational forecasting system.

Description: Published

Description: 1195-1203

Description: JCR Journal

Description: reserved

Description: The accuracy of all types of Vaisala radiosondes and two types of Snow White chilled-mirror hygrosondes was assessed in an intensive in situ comparison with reference hygrometers. Fourteen nighttime reference comparisons were performed to determine a working reference for the radiosonde comparisons. These showed that the night version of the Snow White agreed best with the references [i.e., the NOAA frost-point hygrometer (FPH) and University of Colorado cryogenic frost-point hygrometer (CFH)], but that the daytime version had severe problems with contamination in the humid upper troposphere. Since the RS92 performance was superior to the other radiosondes and to the day version of the Snow White, it was selected to be the working reference. According to the reference comparison, the RS92 has no bias in the mid- and lower troposphere, with deviations 〈±5% in relative humidity (RH). In the upper troposphere, the RS92 has a 5% RH wet bias, which is partly due to the RS92 time lag error and the termination of the heating cycle. It was shown that the time lag effects relating to Vaisala radiosondes can be corrected. Because these were nighttime comparisons, they can be considered to be free from solar radiation effects. Neither the radiosondes nor the Snow White succeeded in reproducing reference class hygrometer profiles in the stratosphere. According to the 29 radiosonde intercomparisons, the RS92 and the modified RS90 (FN) had the best mutual agreement and no bias. The disagreement is largest (〈±10% RH) at low temperatures (T ≪ −30°C), where the FN underestimated (overestimated) in high (low) ambient RH. In comparison with the RS92, the RS90 had a semilinearly increasing wet bias with decreasing temperature, where the bias was 10% RH at −60°C. The RS80-A suffers from a large temperature-dependent dry bias in high RH conditions, being over 30% RH at −60°C and 5% RH near 0°C. The RS80-A dry bias can be almost totally removed with the correction algorithm by Leiterer et al., which was chosen as the best available. The other approach tested tends to overcorrect in high RH conditions when T 〈 −50°C. For T 〉 −30°C it is ineffective and does not correct the RS80-A dry bias in high ambient RH.

Description: In this study we show a teleconnection pattern relating Outgoing Longwave Radiation (OLR) anomalies over the western Pacific Ocean and sea surface temperature anomalies (SSTA) over the western Indian Ocean over two seasons (Sept-Oct-Nov and Dec-Jan-Feb) at zero lag from observations and atmospheric general circulation model (AGCM) integrations. This teleconnection pattern suggests that a positive SSTA in Sept-Oct-Nov (SON) and Dec-Jan-Feb (DJF) seasons over the western Indian Ocean increases the contemporaneous positive OLR anomalies over the western Pacific Ocean. This teleconnection pattern is also simulated by the Center for Ocean-Land- Atmosphere studies (COLA) AGCM forced with observed SST’s. From the experimental COLA AGCM runs (wherein the Pacific Ocean SST variability is suppressed except for the climatological annual cycle) it is diagnosed that the interannual variability of OLR over the western Pacific Ocean persists because of this teleconnection. In relation to this teleconnection pattern it is shown that there is a significant linear response of the SON and DJF equatorial zonal wind anomaly over the Pacific Ocean to contemporaneous SSTA over the western Indian Ocean which is comparable to that of the eastern and western Pacific Oceans. The experimental AGCM runs clearly show that this response of the equatorial zonal wind anomaly to the western Indian Ocean forcing shifts westward towards the Indian Ocean in the absence of Pacific SST variability.

Description: Published

Description: Skill in ensemble-mean dynamical seasonal climate hindcasts with a coupled land-atmosphere model and specified observed sea surface temperature is compared to that for long multi-decade integrations of the same model where the initial conditions are far removed from the seasons of validation. The evaluations are performed for surface temperature and compared among all seasons. Skill is found to be higher in the seasonal simulations than the multi-decadal integrations except during boreal winter. The higher skill is prominent even beyond the first month when the direct influence of the atmospheric initial state elevates model skill. Skill is generally found to be lowest during the winter season for the dynamical seasonal forecasts, equal to that of the long integrations, which show some of the highest skill during winter. The reason for the differences in skill during the non-winter months is attributed to the severe climate drift in the long simulations, manifest through errors in downward fluxes of water and energy over land and evident in soil wetness. The drift presses the land surface to extreme dry or wet states over much of the globe, into a range where there is little sensitivity of evaporation to fluctuations in soil moisture. Thus, the land-atmosphere feedback is suppressed, which appears to lessen the model’s ability to respond correctly over land to remote ocean temperature anomalies.

Description: Center for Ocean-Land-Atmosphere Studies

Description: Published

Description: In this paper, the circulations driven by deep heating and shallow heating are investigated through analytically solving a set of linear equations and examining circulations simulated by a dry primitive equation model. Special emphasis is placed on the low-level mass (moisture) convergence associated with the forced circulation and the maintenance of the shallow and deep heat sources. It is found that the forced circulation driven by shallow heating is more likely to be trapped horizontally near the heating area but relatively extended in the vertical. As a consequence, diabatic heating can not balance adiabatic cooling due to upward motion. At the levels slightly above the top of the heating, a negative vertical gradient of temperature perturbation appears. For the atmosphere driven by deep heating, however, the temperature perturbation cannot accumulate because the heating signals propagate away very fast, allowing an approximate equilibrium between the convective diabatic heating and adiabatic cooling due to upward motion. The converged moisture associated with circulation driven by shallow heating exceeds the amount needed to maintain the heat source. However, the circulation driven by deep heating does not feed back effectively to the moisture convergence, and thus can not be self-sustaining.

Description: Center for Ocean-Land-Atmosphere Studies - Calverton

Description: Published

Description: This paper is not subject to U.S. copyright. The definitive version was published in Journal of Atmospheric and Oceanic Technology 21 (2004): 1448–1461, doi:10.1175/1520-0426(2004)021〈1448:AOAPAD〉2.0.CO;2.

Description: The accuracy of velocities measured by a pulse-coherent acoustic Doppler profiler (PCADP) in the bottom boundary layer of a wave-dominated inner-shelf environment is evaluated. The downward-looking PCADP measured velocities in eight 10-cm cells at 1 Hz. Velocities measured by the PCADP are compared to those measured by an acoustic Doppler velocimeter for wave orbital velocities up to 95 cm s−1 and currents up to 40 cm s−1. An algorithm for correcting ambiguity errors using the resolution velocities was developed. Instrument bias, measured as the average error in burst mean speed, is −0.4 cm s−1 (standard deviation = 0.8). The accuracy (root-mean-square error) of instantaneous velocities has a mean of 8.6 cm s−1 (standard deviation = 6.5) for eastward velocities (the predominant direction of waves), 6.5 cm s−1 (standard deviation = 4.4) for northward velocities, and 2.4 cm s−1 (standard deviation = 1.6) for vertical velocities. Both burst mean and root-mean-square errors are greater for bursts with ub ≥ 50 cm s−1. Profiles of burst mean speeds from the bottom five cells were fit to logarithmic curves: 92% of bursts with mean speed ≥ 5 cm s−1 have a correlation coefficient R2 〉 0.96. In cells close to the transducer, instantaneous velocities are noisy, burst mean velocities are biased low, and bottom orbital velocities are biased high. With adequate blanking distances for both the profile and resolution velocities, the PCADP provides sufficient accuracy to measure velocities in the bottom boundary layer under moderately energetic inner-shelf conditions.

Description: This work was funded by the U.S. Geological Survey as part of the Southwest Washington Coastal Erosion Study

Description: No Abstract available.

Description: Advances in computer power, new forecasting challenges, and new diagnostic techniques have brought about changes in the way atmospheric development and vertical motion are diagnosed in an operational setting. Many of these changes, such as improved model skill, model resolution, and ensemble forecasting, have arguably been detrimental to the ability of forecasters to understand and respond to the evolving atmosphere. The use of nondivergent wind in place of geostrophic wind would be a step in the right direction, but the advantages of potential vorticity suggest that its widespread adoption as a diagnostic tool on the west side of the Atlantic is overdue. Ertel potential vorticity (PV), when scaled to be compatible with pseudopotential vorticity, is generally similar to pseudopotential vorticity, so forecasters accustomed to quasigeostrophic reasoning through the height tendency equation can transfer some of their intuition into the Ertel-PV framework. Indeed, many of the differences between pseudopotential vorticity and Ertel potential vorticity are consequences of the choice of definition of quasigeostrophic PV and are not fundamental to the quasigeostrophic system. Thus, at its core, PV thinking is consistent with commonly used quasigeostrophic diagnostic techniques.

Description: Synoptic and mesoscale meteorology underwent a revolution in the 1940s and 1950s with the widespread deployment of novel weather observations, such as the radiosonde network and the advent of weather radar. These observations provoked a rapid increase in our understanding of the structure and dynamics of the atmosphere by pioneering analysts such as Fred Sanders. The authors argue that we may be approaching an analogous revolution in our ability to study the structure and dynamics of atmospheric phenomena with the advent of probabilistic objective analyses. These probabilistic analyses provide not only best estimates of the state of the atmosphere (e.g., the expected value) and the uncertainty about this state (e.g., the variance), but also the relationships between all locations and all variables at that instant in time. Up until now, these relationships have been determined by sampling in time by, for example, case studies, composites, and time-series analysis. Here the authors propose a new approach, ensemble synoptic analysis, which exploits the information contained in probabilistic samples of analyses at one or more instants in time. One source of probabilistic analyses is ensemble-based state-estimation methods, such as ensemble-based Kalman filters. Analyses from such a filter may be used to study atmospheric phenomena and the relationships between fields and locations at one or more instants in time. After a brief overview of a research-based ensemble Kalman filter, illustrative examples of ensemble synoptic analysis are given for an extratropical cyclone, including relationships between the cyclone minimum sea level pressure and other synoptic features, statistically determined operators for potential-vorticity inversion, and ensemble-based sensitivity analysis.

Description: The pioneering large-scale studies of cyclone frequency, location, and intensity conducted by Fred Sanders prompt similar questions about lesser-studied anticyclone development. The results of a climatology of closed anticyclones (CAs) at 200, 500, and 850 hPa, with an emphasis on the subtropics and midlatitudes, is presented to assess the seasonally varying distribution and hemispheric differences of these features. To construct the CA climatology, a counting program was applied to twice-daily 2.5° NCEP–NCAR reanalysis 200-, 500-, and 850-hPa geopotential height fields for the period 1950–2003. Stationary CAs, defined as those CAs that were located at a particular location for consecutive time periods, were counted only once. The climatology results show that 200-hPa CAs occur preferentially during summer over subtropical continental regions, while 500-hPa CAs occur preferentially over subtropical oceans in all seasons and over subtropical continents in summer. Conversely, 850-hPa CAs occur preferentially over oceanic regions beneath upper-level midocean troughs, and are most prominent in the Northern Hemisphere, and over midlatitude continents in winter. Three case studies of objectively identified CAs that produced heal waves over the United States, Europe, and Australia in 1995, 2003, and 2004, respectively, are presented to supplement the climatological results. The case studies, examining the subset of CAs than can produce heat waves, illustrate how climatologically hot continental tropical air masses produced over arid and semiarid regions of the subtropics and lower midlatitudes can become abnormally hot in conjunction with dynamically driven upper-level ridge amplification. Subsequently, these abnormally hot air masses are advected downstream away from their source regions in conjunction with transient disturbances embedded in anomalously strong westerly jets.

Description: Oklahoma Mesonetwork data are used to illustrate important atmospheric features that are not well shown by the usual synoptic data. For example, some shifts of wind from south to north that are shown as cold fronts on synoptic charts are not cold fronts by any plausible definition. As previously discussed by Fred Sanders and others, such errors in analysis can be reduced by knowledge of the wide variety of weather phenomena that actually exists, and by more attention to temperatures at the earth's surface as revealed by conventional synoptic data. Mesoscale data for four cases reinforce previous discussions of the ephemeral nature of fronts and deficiencies in the usual analyses of cold fronts. One type of misanalyzed case involves post-cold-frontal boundary layer air that is warmer than the prefrontal air. A second type is usually nocturnal, with a rise of local temperature during disruption of an inversion and a wind shift with later cooling that accompanies advection of a climatological gradient of temperature.

Description: The advent of the polar front theory of cyclones in Norway early in the last century held that the development of fronts and air masses is central to understanding midlatitude weather phenomena. While work on fronts continues to this day, the concept of air masses has been largely forgotten, superseded by the idea of a continuum. The Norwegians placed equal emphasis on the thermodynamics of airmass formation and on the dynamical processes that moved air masses around; today, almost all the emphasis is on dynamics, with little published literature on diabatic processes acting on a large scale. In this essay, the author argues that a lack of understanding of large-scale diabatic processes leads to an incomplete picture of the atmosphere and contributes to systematic errors in medium- and long-range weather forecasts. At the same time, modern concepts centered around potential vorticity conservation and inversion lead one to a redefinition of the term "air mass" that may have some utility in conceptualizing atmospheric physics and in weather forecasting.

Description: Historically, the atmospheric sciences have tended to treat problems of weather and climate separately. The real physical system, however, is a continuum, with short-term (minutes to days) “weather” fluctuations influencing climate variations and change, and, conversely, more slowly varying aspects of the system (typical time scales of a season or longer) affecting the weather that is experienced. While this past approach has served important purposes, it is becoming increasingly apparent that in order to make progress in addressing many socially important problems, an improved understanding of the connections between weather and climate is required. This overview summarizes the progress over the last few decades in the understanding of the phenomena and mechanisms linking weather and climate variations. The principal emphasis is on developments in understanding key phenomena and processes that bridge the time scales between synoptic-scale weather variability (periods of approximately 1 week) and climate variations of a season or longer. Advances in the ability to identify synoptic features, improve physical understanding, and develop forecast skill within this time range are reviewed, focusing on a subset of major, recurrent phenomena that impact extratropical wintertime weather and climate variations over the Pacific–North American region. While progress has been impressive, research has also illuminated areas where future gains are possible. This article concludes with suggestions on near-term directions for advancing the understanding and capabilities to predict the connections between weather and climate variations.

Description: Fred Sanders' career extended over 55 yr, touching upon many of the revolutionary transformations in the field of meteorology during that period. In this paper, his contributions to the transformation of synoptic meteorology, his research into the nature of explosive cyclogenesis, and related advances in the ability to predict these storms are reviewed. In addition to this review, the current status of forecasting oceanic cyclones 4.5 days in advance is presented, illustrating the progress that has been made and the challenges that persist, especially for forecasting those extreme extratropical cyclones that are marked by surface wind speeds exceeding hurricane force. Last, Fred Sanders' participation in a forecast for the historic 1947 snowstorm (that produced snowfall amounts in the New York City area that set records at that time) is reviewed along with an attempt to use today's operational global model to simulate this storm using data that were available at the time. The study reveals the predictive limitations involved with this case based on the scarcity of upper-air data in 1947, while confirming Fred Sanders' forecasting skills when dealing with these types of major storm events, even as a young aviation forecaster at New York's LaGuardia Airport.

Description: A case study of a double dryline on 22 May 2002 is presented. Mobile, 3-mm-wavelength Doppler radars from the University of Massachusetts and the University of Wyoming (Wyoming cloud radar) were used to collect very fine resolution vertical-velocity data in the vicinity of each of the moisture gradients associated with the drylines. Very narrow (50–100 m wide) channels of strong upward vertical velocity (up to 8 m s–1) were measured in the convergence zone of the easternmost dryline, larger in magnitude than reported with previous drylines. Distinct areas of descending motion were evident to the east and west of both drylines. Radar data are interpreted in the context of other observational platforms available during the International H2O Project (IHOP-2002). a variational ground-based mobile radar data processing technique was developed and applied to pseudo-dual-Doppler data collected during a rolling range-height indicator deployment. It was found that there was a secondary (vertical) circulation normal to the easternmost moisture gradient; the circulation comprised an easterly component near-surface flow to the east, a strong upward vertical component in the convergence zone, a westerly return, flow above the convective boundary layer, and numerous regions of descending motion, the most prominent approximately 3–5 km to the east of the surface convergence zone.

Description: Fred Sanders' teaching and research contributions in the area of quasigeostrophic theory are highlighted in this paper. The application of these contributions is made to the topic of extreme cold-season precipitation events in the Saint Lawrence valley in the northeastern United States and southern Quebec. This research focuses on analyses of Saint Lawrence valley heavy precipitation events. Synoptic- and planetary-scale circulation anomaly precursors are typically identified several days prior to these events. These precursors include transient upper-level troughs, strong moisture transports into the region, and anomalously large precipitable water amounts. The physical insight of Fred Sanders' work is used in the analysis of these composite results. Further details of this insight are provided in analyses of one case of heavy precipitation.

Description: Sanders designed a barotropic tropical cyclone (TC) track prediction model for the North Atlantic TC basin that became known as the Sanders barotropic (SANBAR) model. It predicted the streamfunction of the deeplayer mean winds (tropical circulation vertically averaged from 1000 to 100 hPa) that represents the vertically averaged tropical circulations. Originally, the wind input for the operational objective analysis (OA) consisted of winds measured by radiosondes and 44 bogus winds provided by analysis at the National Hurricane Center (NHC), which corresponded to the vertically averaged flow over sparsely observed tropical, subtropical, and midlatitude oceanic regions. The model covered a fixed regional area and had a grid size of ~ 154 km. It estimated the initial storm motion solely on the basis of the large-scale flow from the OA, not taking into account the observed storm motion. During 1970, the SANBAR model became the first dynamical TC track model to be run operationally at NHC. Track forecasts of SANBAR were verified from the 1971 TC season when track model verifications began at NHC until its retirement after the 1989 Atlantic TC season. The average annual SANBAR forecast track errors were verified relative to Climatology and Persistence (CLIPER), the standard no-skill track forecast. Comparison with CLIPER determines the skill of track forecast methods. Verifications are presented for two different versions of the SANBAR model system used operationally during 1973–84 and 1985–89. In homogeneous comparisons (i.e., includes only forecasts for the same initial times) for the former period, SANBAR's track forecasts were slightly better than CLIPER at 24–48-h forecast intervals; however, from 1985 to 1989 the average SANBAR track forecast errors from 24–72 h were ~10% more skillful than homogeneous CLIPER track forecasts.

Description: In the last decade, Fred Sanders was often critical of current surface analysis techniques. This led to his promoting the use of surface potential temperatures to distinguish between fronts, baroclinic troughs, and non-frontal baroclinic zones, and to the development of a climatology of surface baroclinic zones. In this paper, criticisms of current surface analysis techniques and the usefulness of surface potential temperature analyses are discussed. Case examples are used to compare potential temperature analyses and current National Centers for Environmental Prediction analyses. The 1-yr climatology of Sanders and Hoffman is reconstructed using a composite technique. Annual and seasonal mean potential temperature analyses over the continental United States, southern Canada, northern Mexico, and adjacent coastal waters are presented. In addition, gridpoint frequencies of moderate and strong potential temperature gradients are calculated. The results of the mean potential temperature analyses show that moderate and strong surface baroclinic zones are favored along the coastlines and the slopes of the North American cordillera. Additional subsynoptic details, not found in Sanders and Hoffman, are identified. The availability of the composite results allows for the calculation of potential temperature gradient anomalies. It is shown that these anomalies can be used to identify significant frontal baroclinic zones that are associated with weak potential temperature gradients. Together the results and reviews in this paper show that surface potential temperature analyses are a valuable forecasting and analysis tool allowing analysts to distinguish and identify fronts, baroclinic troughs, and nonfrontal baroclinic zones.

Description: The nature of the different types of surface boundaries that appear in the southern plains of the United States during the convectively active season is reviewed. The following boundaries are discussed: fronts, the dryline, troughs, and outflow boundaries, The boundaries are related to their environment and to local topography. The role these boundaries might play in the initiation of convective storms is emphasized. The various types of boundary-related vertical circulations and their dynamics are discussed. In particular, quasigeostrophic and semigeostrophic dynamics, and the dynamics of solenoidal circulations, density currents, boundary layers, and gravity waves are considered. Miscellaneous topics pertinent to convective storms and their relationship to surface boundaries such as along-the-boundary variability, boundary collisions, and the role of vertical shear are also discussed. Although some cases of storm initiation along surface boundaries have been well documented using research datasets collected during comprehensive field experiments, much of what we know is based only on empirical forecasting and nowcasting experience. It is suggested that many problems relating to convective-storm formation need to be explored in detail using real datasets with new observing systems and techniques, in conjunction with numerical simulation studies, and through climatological studies.

Description: This paper begins with a review of basic surface frontogenesis concepts with an emphasis on fronts located over sloping terrain adjacent to mountain barriers and fronts located in large-scale baroclinic zones close to coastlines. The impact of cold-air damming and differential diabatic heating and cooling on frontogenesis is considered through two detailed case studies of intense surface fronts. The first case, from 17 to 18 April 2002, featured the westward passage of a cold (side-door) front across coastal eastern New England in which 15°–20°C temperature decreases were observed in less than one hour. The second case, from 28 February to 4 March 1972, featured a long-lived front that affected most of the United States from the Rockies to the Atlantic coast and was noteworthy for a 50°C temperature contrast between Kansas and southern Manitoba, Canada. In the April 2002 case most of New England was initially covered by an unusually warm, dry air mass. Dynamical anticyclogenesis over eastern Canada set the stage for a favorable pressure gradient to allow chilly marine air to approach coastal New England from the east. Diabatic cooling over the chilly (5°–8°C) waters of the Gulf of Maine allowed surface pressures to remain relatively high offshore while diabatic heating over the land (31°–33°C temperatures) enabled surface pressures to fall relative to over the ocean. The resulting higher pressures offshore resulted in an onshore cold push. Frontal intensity was likely enhanced prior to leaf out and grass green-up as virtually all of the available insolation went into sensible heating. The large-scale environment in the February–March 1972 case favored the accumulation of bitterly cold arctic air in Canada. Frontal formation occurred over northern Montana and North Dakota as the arctic air moved slowly southward in conjunction with surface pressure rises east of the Canadian Rockies. The arctic air accelerated southward subsequent to lee cyclogenesis–induced pressure falls ahead of an upstream trough that crossed the Rockies. The southward acceleration of the arctic air was also facilitated by dynamic anticyclogenesis in southern Canada beneath a poleward jet-entrance region. Frontal intensity varied diurnally in response to differential diabatic heating. Three types of cyclogenesis events were observed over the lifetime of the event: 1) low-amplitude frontal waves with no upper-level support, 2) low-amplitude frontal waves that formed in a jet-entrance region, and 3) cyclones that formed ahead of advancing upper-level troughs. All cyclones were either nondeveloping or weak developments despite extreme baroclinicity, likely the result of large atmospheric static stability in the arctic frontal zone and unfavorable alongfront stretching deformation. Significant frontal–mountain interactions were observed over the Rockies and the Appalachians.

Description: Over 50 yr have passed since the publication of Sanders' 1955 study, the first quantitative study of the structure and dynamics of a surface cold front. The purpose of this chapter is to reexamine some of the results of that study in light of modern methods of numerical weather prediction and diagnosis. A simulation with a resolution as high as 6-km horizontal grid spacing was performed with the fifth-generation-Pennsylvania State University-National Center for Atmospheric Research (PSU-NCAR) Mesoscale Model (MM5), given initial and lateral boundary conditions from the National Centers for Environmental Precipitation-National Center for Atmospheric Research (NCEP-NCAR) reanalysis project data from 17 to 18 April 1953. The MM5 produced a reasonable simulation af the front, albeit its strength was not as intense and its movement was not as fast as was analyzed by Sanders. The vertical structure of the front differed from that analyzed by Sanders in several significant ways. First, the strongest horizontal temperature gradient associated with the cold front in the simulation occurred above a surface-based inversion, not at the earth's surface. Second, the ascent plume at the leading edge of the front was deeper and more intense than that analyzed by Sanders. The reason was an elevated mixed layer that had moved over the surface cold front in the simulation, allowing a much deeper vertical circulation than was analyzed by Sanders. This structure is similar to that of Australian cold fronts with their deep, well-mixed, prefrontal surface layer. These two differences between the model simulation and the analysis by Sanders may be because upper-air data from Fort Worth, Texas, was unavailable to Sanders. Third, the elevated mixed layer also meant that isentropes along the leading edge of the front extended vertically. Fourth, the field of frontogenesis of the horizontal temperature gradient calculated from the three-dimensional wind differed in that the magnitude of the maximum of the deformation term was larger than the magnitude of the maximum of the tilting term in the simulation, in contrast to Sanders' analysis and other previously published cases. These two discrepancies may be attributable to the limited horizontal resolution of the data that Sanders used in constructing his cross section. Last, a deficiency of the model simulation was that the postfrontal surface superadiabatic layer in the model did not match the observed well-mixed boundary layer. This result raises the question of the origin of the well-mixed postfrontal boundary layer behind cold fronts. To address this question, an additional model simulation without surface fluxes was performed, producing a well-mixed, not superadiabatic, layer. This result suggests that surface fluxes were not necessary for the development of the well-mixed layer, in agreement with previous research. Analysis of this event also amplifies two research themes that Sanders returned to later in his career, First, a prefrontal wind shift occurred in both the observations and model simulation at stations in western Oklahoma. This prefrontal wind shift was caused by a lee cyclone departing the leeward slopes of the Rockies slightly equatorward of the cold front, rather than along the front as was the case farther eastward. Sanders' later research showed how the occurrence of these prefrontal wind shifts leads to the weakening of fronts. Second, this study shows the advantage of using surface potential temperature, rather than surface temperature, for determining the locations of the surface fronts on sloping terrain.

Description: Today, even with state-of-the-art observational, data assimilation, and modeling systems run routinely on supercomputers, there are often surprises in the prediction of snowstorms, especially the “big ones,” affecting coastal regions of the mid-Atlantic and northeastern United States. Little did the author know that lessons from Fred Sanders' synoptic meteorology class at the Massachusetts Institute of Technology (1967) would later (late 1980s) inspire him to pursue practical issues of predictability in the context of the development of ensemble prediction systems, strategies, and applications for providing information on the inevitable case-dependent uncertainties in forecasts. This paper is a brief qualitative and somewhat colloquial overview, based upon this author's personal involvement and experiences, intended to highlight some basic aspects of the source and nature of uncertainties in forecasts and to illustrate the sort of value added information ensembles can provide in dealing with uncertainties in predictions of East Coast snowstorms.

Description: One characteristic of Fred Sanders' research is his ability to take a topic that is believed to be well understood by the research community and show that interesting research problems still exist. Among Sanders' considerable contributions to synoptic meteorology, those concerned with surface cold fronts have been especially influential. After a brief historical review of fronts and frontal analysis, this chapter presents three stages in Sanders' career when he performed research on the structure, dynamics, and analysis of surface cold fronts. First, his 1955 paper, "An investigation of the structure and dynamics of an intense surface frontal zone," was the first study to discuss quantitatively the dynamics of a surface cold front. In the 1960s, Sanders and his students further examined the structure of cold fronts, resulting in the unpublished 1967 report to the National Science Foundation, "Frontal structure and the dynamics of frontogenesis." For a third lime in his career, Sanders published several papers (1995–2005) revisiting the structure and dynamics of cold fronts. His 1967 and 1995–2005 work raises the question of the origin and dynamics of the surface pressure trough and/or wind shift that sometimes precedes the temperature gradient (hereafter called a prefrontal trough or prefrontal wind shift, respectively). Sanders showed that the relationship between this prefrontal feature and the temperature gradient is fundamental to the strength of the front. When the wind shift is coincident with the temperature gradient, frontogenesis (strengthening of the front) results; when the wind shift lies ahead of the temperature gradient, frontolysis (weakening of the front) results. a number of proposed mechanisms for the formation of prefrontal troughs and prefrontal wind shifts exist. Consequently, much research remains to be performed on these topics.

Description: No Abstract available.

Description: When a forecast is assessed, a single value for a verification measure is often quoted. This is of limited use, as it needs to be complemented by some idea of the uncertainty associated with the value. If this uncertainty can be quantified, it is then possible to make statistical inferences based on the value observed. There are two main types of inference: confidence intervals can be constructed for an underlying “population” value of the measure, or hypotheses can be tested regarding the underlying value. This paper will review the main ideas of confidence intervals and hypothesis tests, together with the less well known “prediction intervals,” concentrating on aspects that are often poorly understood. Comparisons will be made between different methods of constructing confidence intervals—exact, asymptotic, bootstrap, and Bayesian—and the difference between prediction intervals and confidence intervals will be explained. For hypothesis testing, multiple testing will be briefly discussed, together with connections between hypothesis testing, prediction intervals, and confidence intervals.

Description: Studies using International Satellite Cloud Climatology Project (ISCCP) data have reported decreases in cloud optical depth with increasing temperature, thereby suggesting a positive feedback in cloud optical depth as climate warms. The negative cloud optical depth and temperature relationships are questioned because ISCCP employs threshold assumptions to identify cloudy pixels that have included partly cloudy pixels. This study applies the spatial coherence technique to one month of Advanced Very High Resolution Radiometer (AVHRR) data over the Pacific Ocean to differentiate overcast pixels from the partly cloudy pixels and to reexamine the cloud optical depth–temperature relationships. For low-level marine stratus clouds studied here, retrievals from partly cloudy pixels showed 30%–50% smaller optical depths, 1°–4°C higher cloud temperatures, and slightly larger droplet effective radii, when they were compared to retrievals from the overcast pixels. Despite these biases, retrievals for the overcast and partly cloudy pixels show similar negative cloud optical depth–temperature relationships and their magnitudes agree with the ISCCP results for the midlatitude and subtropical regions. There were slightly negative droplet effective radius–temperature relationships, and considerable positive cloud liquid water content–temperature relationships indicated by aircraft measurements. However, cloud thickness decreases appear to be the main reason why cloud optical depth decreases with increasing temperature. Overall, cloud thickness thinning may explain why similar negative cloud optical depth–temperature relationships are found in both overcast and partly cloudy pixels. In addition, comparing the cloud-top temperature to the air temperature at 740 hPa indicates that cloud-top height generally rises with warming. This suggests that the cloud thinning is mainly due to the ascending of cloud base. The results presented in this study are confined to the midlatitude and subtropical Pacific and may not be applicable to the Tropics or other regions.

Description: A statistical model to analyze different time scales of the variability of extreme high sea levels is presented. This model uses a time-dependent generalized extreme value (GEV) distribution to fit monthly maxima series and is applied to a large historical tidal gauge record (San Francisco, California). The model allows the identification and estimation of the effects of several time scales—such as seasonality, interdecadal variability, and secular trends—in the location, scale, and shape parameters of the probability distribution of extreme sea levels. The inclusion of seasonal effects explains a large amount of data variability, thereby allowing a more efficient estimation of the processes involved. Significant correlation with the Southern Oscillation index and the nodal cycle, as well as an increase of about 20% for the secular variability of the scale parameter have been detected for the particular dataset analyzed. Results show that the model is adequate for a complete analysis of seasonal-to-interannual sea level extremes providing time-dependent quantiles and confidence intervals.

Description: The interannual and intraseasonal variability of West African vegetation over the period 1982–2002 is studied using the normalized difference vegetation index (NDVI) from the Advanced Very High Resolution Radiometer (AVHRR).The novel independent component analysis (ICA) technique is applied to extract the main modes of the interannual variability of the vegetation, among which two modes are worth describing. The first component (IC1) describes NDVI variability over the Sahel from August to October. A strong photosynthetic activity over the Sahel is related to above-normal convection and rainfall within the intertropical convergence zone (ITCZ) in summertime and is partly associated with colder (warmer) SST in the eastern tropical Pacific (the Mediterranean). The second component (IC2) depicts a dipole pattern between the Sahelian and Guinean regions during the northern summer followed by a southward-propagating signal from October to December. It is associated with a north–south dipole in convection and rainfall induced by variations in the latitudinal location of the ITCZ as a response to the occurrence of the tropical Atlantic dipole.The analysis of the intraseasonal variability of the Sahelian vegetation relies on the analysis of the seasonal marches and their main phenological stages. Green-up usually starts in early July and shows a very low year-to-year variability, while senescence ends by mid-November and is prone to larger interannual variability. Six types of vegetative seasonal marches are discriminated according to variations in the timing of phenological stages as well as in the greening intensity. These types appear to be strongly dependent on rainfall distribution and amount, particularly those recorded in late August. Finally, year-to-year memory effects are highlighted: NDVI recorded during the green-up phase in year j appears to be strongly related to the maximum NDVI value recorded at year j − 1.

Description: Based on the data of optimum interpolation sea surface temperature (OISST), the temporal correlations of the sea surface temperature anomaly (SSTA) in the South China Sea (SCS) are studied by using the rescaled range analysis (R/S) and detrended fluctuation analysis (DFA). The results show that the scaling exponents of SSTAs are larger than 0.8. This finding indicates that the SSTAs in the SCS exhibit persistent long-range time correlation of the fluctuations and the interval spreads over a wide period, from about 1 month to 4.5 yr (4∼235 weeks). In addition, the “degree” of the correlations depends very much on the geographic locations: near to the coastal regions, the value is small, while far from the coastline, the value is relatively larger. This means that SSTAs in the central SCS are smoother than those of the coastal regions. The persistence of SST in the SCS may be used as a “minimum skill” to assess the ocean models and to evaluate their performance.

Description: Winds at the Salt Lake City International Airport (SLC) during the April–October period from 1948 to 2003 have been observed to shift to the north (up-valley direction) between late morning and afternoon on over 70% of the days without precipitation. Lake-breeze fronts that develop as a result of the differential heating between the air over the nearby Great Salt Lake and that over the lake’s surroundings are observed at SLC only a few times each month. Fewer lake-breeze fronts are observed during late July–early September than before or after that period. Interannual fluctuations in the areal extent of the shallow Great Salt Lake contribute to year-to-year variations in the number of lake-breeze frontal passages at SLC. Data collected during the Vertical Transport and Mixing Experiment (VTMX) of October 2000 are used to examine the structure and evolution of a lake-breeze front that moved through the Salt Lake Valley on 17 October. The onset of upslope and up-valley winds occurred within the valley prior to the passage of the lake-breeze front. The lake-breeze front moved at roughly 3 m s−1 up the valley and was characterized near the surface by an abrupt increase in wind speed and dewpoint temperature over a distance of 3–4 km. Rapid vertical mixing of aerosols at the top of the 600–800-m-deep boundary layer was evident as the front passed.

Description: This study combines the experimental measurements with large-eddy simulation (LES) data of a neutral planetary boundary layer (PBL) documented by a 60-m tower instrumented with eight sonic anemometers, and a high-resolution Doppler lidar during the 1999 Cooperative Atmospheric and Surface Exchange Study (CASES-99) experiment. The target of the paper is (i) to investigate the multiscale nature of the turbulent eddies in the surface layer (SL), (ii) to explain the existence of a −1 power law in the velocity fluctuation spectra, and (iii) to investigate the different nature of turbulence in the two sublayers within the SL, which are the eddy surface layer (ESL; lower sublayer of the SL lying between the surface and about 20-m height) and the shear surface layer (SSL; lying between the ESL top and the SL top). The sonic anemometers and Doppler lidar prove to be proper instruments for LES validation, and in particular, the Doppler lidar because of its ability to map near-surface eddies.This study shows the different nature of turbulence in the ESL and the SSL in terms of organized eddies, velocity fluctuation spectra, and second-order moment statistics. If quantitative agreement is found in the SSL between the LES and the measurements, only qualitative similarity is found in the ESL due to the subgrid-scale model, indicating that the LES captures part of the physics of the ESL. The LES helps explain the origin of the −1 power-law spectral subrange evidence in the velocity fluctuation spectra computed in the SL using the CASES-99 dataset: strong interaction between the mean flow and the fluctuating vorticities is found in the SL, and turbulent production is found to be larger than turbulent energy transfer for k1z 〉 1 (k1 being the longitudinal wavenumber and z the height), which is the condition of the −1 power-law existence.

Description: The effects of natural and anthropogenic heterogeneity on a hydrological simulation are evaluated using a distributed biosphere hydrological model (DBHM) system. The DBHM embeds a biosphere model into a distributed hydrological scheme, representing both topography and vegetation in a mesoscale hydrological simulation, and the model system includes an irrigation scheme. The authors investigated the effects of two kinds of variability, precipitation variability and the variability of irrigation redistributing runoff, representing natural and anthropogenic heterogeneity, respectively, on hydrological processes. Runoff was underestimated if rainfall was placed spatially uniformly over large grid cells. Accounting for precipitation heterogeneity improved the runoff simulation. However, the negative runoff contribution, namely, the situation that mean annual precipitation is less than evapotranspiration, cannot be simulated by only considering the natural heterogeneity. This constructive model shortcoming can be eliminated by accounting for anthropogenic heterogeneity caused by irrigation water withdrawals. Irrigation leads to increased evapotranspiration and decreased runoff, and surface soil moisture in irrigated areas increases because of irrigation. Simulations performed for the Yellow River basin of China indicated streamflow decreases of 41% due to irrigation effects. The latent heat flux in the peak irrigation season [June–August (JJA)] increased 3.3 W m−2 with a decrease in the ground surface temperature of 0.1 K for the river basin. The maximum simulated increase in the latent heat flux was 43 W m−2, and the ground temperature decrease was 1.6 K in the peak irrigation season.

Description: A method for routinely verifying numerical weather prediction surface marine winds with satellite scatterometer winds is introduced. The marine surface winds from the Australian Bureau of Meteorology’s operational global and regional numerical weather prediction systems are evaluated. The model marine surface layer is described. Marine surface winds from the global and limited-area models are compared with observations, both in situ (anemometer) and remote (scatterometer). A 2-yr verification shows that wind speeds from the regional model are typically underestimated by approximately 5%, with a greater bias in the meridional direction than the zonal direction. The global model also underestimates the surface winds by around 5%–10%. A case study of a significant marine storm shows that where larger errors occur, they are due to an underestimation of the storm intensity, rather than to biases in the boundary layer parameterizations.

Description: The applicability of axisymmetric theory of angular momentum conserving circulations to the large-scale steady monsoon is studied in a general circulation model with idealized representations of continental geometry and simple physics. Results from an aquaplanet setup with localized subtropical forcing are compared with a continental case. It is found that the meridional circulation that develops is close to angular momentum conserving for cross-equatorial circulation cells, both in the aquaplanet and in the continental cases. The equator proves to be a substantial barrier to boundary layer meridional flow; flow into the summer hemisphere from the winter hemisphere tends to occur in the free troposphere rather than in the boundary layer. A theory is proposed to explain the location of the monsoon; assuming quasiequilibrium, the poleward boundary of the monsoon circulation is collocated with the maximum in subcloud moist static energy, with the monsoon rains occurring near and slightly equatorward of this maximum. The model results support this theory of monsoon location, and it is found that the subcloud moist static energy distribution is determined by a balance between surface forcing and advection by the large-scale flow.

Description: This paper concerns the calculation of the probability of exceedance of wave crest elevation. The statistics have been calculated for broadbanded, unidirectional, deep-water sea states by incorporating a fully nonlinear wave model into a spectral response surface method. This is a novel approach to the calculation of statistics and, as all of the calculations are performed in the probability domain, avoids the need for long time-domain simulations. Furthermore, in contrast to theoretical distributions, the broadbanded, fully nonlinear nature of the sea state can be considered. The results have been compared with those of fully nonlinear time-domain simulations and are shown to be in good agreement. The results indicate that in unidirectional seas the crest elevations of the largest waves can be much higher than would be predicted by linear or second-order theory. Hence, the occurrence of locally long crested sea states offers a possible explanation for the formation of freak or rogue waves.

Description: Nearly one-half of the earth’s terrestrial surface is susceptible to drought, which can have significant social, economic, and environmental impacts. Therefore, it is important to develop better descriptions and models of the processes linking the land surface and atmosphere during drought. Using data collected during the International H2O Project, the study presented here investigates the effects of variations in the environmental factors driving the latent heat flux (λE) during drought conditions at a rangeland site located in the panhandle of Oklahoma. Specifically, this study focuses on the relationships of λE with vapor pressure deficit, wind speed, net radiation, soil moisture content, and greenness fraction. While each of these environmental factors has an influence, soil moisture content is the key control on λE. The role of soil moisture in regulating λE is explained in terms of the surface resistance to water vapor transfer. The results show that λE transitioned between being water or energy limited during the course of the drought. The implications of this on the ability to understand and model drought conditions and transitions into or out of droughts are discussed.

Description: Common Land Model (CLM) and Land Surface Process (LSP) model simulations are compared to measured values for a 13-day dry-down period with a rapidly decreasing near-surface water table for a marsh wetland community in Florida. LSP was able to provide reasonable estimates without any modifications to the model physics. To obtain reasonable simulations using CLM, the baseline TOPMODEL baseflow generation and the bottom drainage mechanisms were not employed and the lower layers were allowed to remain saturated. In addition, several of CLM’s default wetland vegetation parameters were replaced with grassland parameters. Even after these modifications, CLM underestimated soil water storage. However, both model-simulated soil temperatures showed very good agreement as compared to measured temperatures, capturing both the soil warming during the study period and the diurnal fluctuations. Modeled surface energy fluxes also agreed well with measured values. LSP’s inability to consistently capture latent heat fluxes appears to be linked to its canopy resistance scaling functions. Other minor issues were that CLM’s rooting depth greatly exceeded observed depths and that CLM did not move water in the vadose zone from lower to upper layers during the nighttime as observed in the measurements. Overall, these results suggest that LSP can be applied to characterize a marsh dry down, but that minor modifications could greatly improve results. CLM demonstrated considerable potential, but requires some changes to model physics and default parameters prior to application to wetlands at a subgrid scale.

Description: In March 2003 several autonomous underwater vehicle (AUV) missions were carried out under sea ice in the western Bellingshausen Sea. Data from the upward-looking acoustic Doppler current profiler (ADCP) on the “Autosub” AUV indicate a strongly oscillating horizontal velocity of the ice due to ocean swell. Swell period, height, direction, and directional spread have been computed every 800 m from the ice edge to 10 km inward for three missions. Exponential, period-dependent attenuation of waves propagating through sea ice was observed. Mean period increased with distance from the ice edge. The wave field refracted during propagation. The directional wave spread does not seem to relate to distance from the ice edge, although higher frequencies tended to be more spread. If suitably deployed, an ordinary ADCP may be used with this technique to study both scalar and directional properties of waves in open or ice-covered water.

Description: Wind-sea generation was observed during two experiments off the coast of North Carolina. One event with offshore winds of 9–11 m s−1 directed 20° from shore normal was observed with eight directional stations recording simultaneously and spanning a fetch from 4 to 83 km. An opposing swell of 1-m height and 10-s period was also present. The wind-sea part of the wave spectrum conforms to established growth curves for significant wave height and peak period, except at inner-shelf stations where a large alongshore wind-sea component was observed. At these short fetches, the mean wave direction θm was observed to change abruptly across the wind-sea spectral peak, from alongshore at lower frequencies to downwind at higher frequencies. Waves from another event with offshore winds of 6–14 m s−1 directed 20°–30° from shore normal were observed with two instrument arrays. A significant amount of low-frequency wave energy was observed to propagate alongshore from the region where the wind was strongest. These measurements are used to assess the performance of some widely used parameterizations in wave models. The modeled transition of θm across the wind-sea spectrum is smoother than that in the observations and is reproduced very differently by different parameterizations, giving insights into the appropriate level of dissipation. Calculations with the full Boltzmann integral of quartet wave–wave interactions reveal that the discrete interaction approximation parameterization for these interactions is reasonably accurate at the peak of the wind sea but overpredicts the directional spread at high frequencies. This error is well compensated by parameterizations of the wind input source term that have a narrow directional distribution. Observations also highlight deficiencies in some parameterizations of wave dissipation processes in mixed swell–wind-sea conditions.

Description: Seven sets of 2D particle image velocimetry data obtained in the bottom boundary layer of the coastal ocean along the South Carolina and Georgia coast [at the South Atlantic Bight Synoptic Offshore Observational Network (SABSOON) site] are examined, covering the accelerating and decelerating phases of a single tidal cycle at several heights above the seabed. Additional datasets from a previous deployment are also included in the analysis. The mean velocity profiles are logarithmic, and the vertical distribution of Reynolds stresses normalized by the square of the free stream velocity collapse well for data obtained at the same elevation but at different phases of the tidal cycle. The magnitudes of 〈u′u′〉, 〈w′w′〉, and −〈u′w′〉 decrease with height above bottom in the 25–160-cm elevation range and are consistent with the magnitudes and trends observed in laboratory turbulent boundary layers. If a constant stress layer exists, it is located below 25-cm elevation. Two methods for estimating dissipation rate are compared. The first, a direct estimate, is based on the measured in-plane instantaneous velocity gradients. The second method is based on fitting the resolved part of the dissipation spectrum to the universal dissipation spectrum available in Gargett et al. Being undervalued, the direct estimates are a factor of 2–2.5 smaller than the spectrum-based estimates. Taylor microscale Reynolds numbers for the present analysis range from 24 to 665. Anisotropy is present at all resolved scales. At the transition between inertial and dissipation range the longitudinal spectra exhibit a flatter than −5/3 slope and form spectral bumps. Second-order statistics of the velocity gradients show a tendency toward isotropy with increasing Reynolds number. Dissipation exceeds production at all measurement heights, but the difference varies with elevation. Close to the bottom, the production is 40%–70% of the dissipation, but it decreases to 10%–30% for elevations greater than 80 cm.

Description: Scalar exchange between San Francisco Bay and the coastal ocean is examined using shipboard observations made across the Golden Gate Channel. The study consists of experiments during each of the following three “seasons”: winter/spring runoff (March 2002), summer upwelling (July 2003), and autumn relaxation (October 2002). Within each experiment, transects across the channel were repeated approximately every 12 min for 25 h during both spring and neap tides. Velocity was measured from a boat-mounted ADCP. Scalar concentrations were measured at the surface and from a tow-yoed SeaSciences Acrobat. Net salinity exchange rates for each season are quantified with harmonic analysis. Accuracy of the net fluxes is confirmed by comparison with independently measured values. Harmonic results are then decomposed into flux mechanisms using temporal and spatial correlations. In this study, the temporal correlation of cross-sectionally averaged salinity and velocity (tidal pumping flux) is the largest part of the dispersive flux of salinity into the bay. From the tidal pumping flux portion of the dispersive flux, it is shown that there is less exchange than was found in earlier studies. Furthermore, tidal pumping flux scales strongly with freshwater flow resulting from the density-driven movement of a tidally trapped eddy and stratification-induced increases in ebb–flood frictional phasing. Complex bathymetry makes salinity exchange scale differently with flow than would be expected from simple tidal asymmetry and gravitational circulation models.

Description: As part of a program aimed at developing a long-duration, subsurface mooring, known as Ultramoor, several modern acoustic current meters were tested. The instruments with which the authors have the most experience are the Aanderaa RCM11 and the Nortek Aquadopp, which measure currents using the Doppler shift of backscattered acoustic signals, and the Falmouth Scientific ACM, which measures changes in travel time of acoustic signals between pairs of transducers. Some results from the Doppler-based Sontek Argonaut and the travel-time-based Nobska MAVS are also reported. This paper concentrates on the fidelity of the speed measurement but also presents some results related to the accuracy of the direction measurement. Two procedures were used to compare the instruments. In one, different instruments were placed close to one another on three different deep-ocean moorings. These tests showed that the RCM11 measures consistently lower speeds than either a vector averaging current meter or a vector measuring current meter, both more traditional instruments with mechanical velocity sensors. The Aquadopp in use at the time, but since updated to address accuracy problems in low scattering environments, was biased high. A second means of testing involved comparing the appropriate velocity component of each instrument with the rate of change of pressure when they were lowered from a ship. Results from this procedure revealed no depth dependence or measurable bias in the RCM11 data, but did show biases in both the Aquadopp and Argonaut Doppler-based instruments that resulted from low signal-to-noise ratios in the clear, low scattering conditions beneath the thermocline. Improvements in the design of the latest Aquadopp have reduced this bias to a level that is not significant.

Description: The energy pathways in geostrophic turbulence are explored using a two-layer, flat-bottom, f-plane, quasigeostrophic model forced by an imposed, horizontally homogenous, baroclinically unstable mean flow and damped by bottom Ekman friction. A systematic presentation of the spectral energy fluxes, the mean flow forcing, and dissipation terms allows for a comprehensive understanding of the sources and sinks for baroclinic and barotropic energy as a function of length scale. The key new result is a robust inverse cascade of kinetic energy for both the baroclinic mode and the upper layer. This is consistent with recent observations of satellite altimeter data over the South Pacific Ocean. The well-known forward cascade of baroclinic potential and total energy was found to be very robust. Decomposing the spectral fluxes into contributions from different terms provided further insight. The inverse baroclinic kinetic energy cascade is driven mostly by an efficient interaction between the baroclinic velocity and the barotropic vorticity, the latter playing a crucial catalytic role. This cascade can be further enhanced by the baroclinic mode self-interaction, which is only present with nonuniform stratification (unequal layer depths). When model parameters are set such that modeled eddies compare favorably with observations, the inverse baroclinic kinetic energy cascade is actually much stronger than the well-known inverse cascade in the barotropic mode. The upper-layer kinetic energy cascade was found to dominate the lower-layer cascade over a wide range of parameters, suggesting that the surface cascade and time mean density stratification may be sufficient for estimating the depth-integrated cascade from ocean observations. This may find useful application in inferring the kinetic to gravitational potential energy conversion rate from satellite measurements.

Description: PU.1, IKAROS, E2A, EBF, and PAX5 comprise a transcriptional network that orchestrates B-cell lineage specification, commitment, and differentiation. Here we identify interferon regulatory factor 8 (IRF8) as another component of this complex, and show that it also modulates lineage choice by hematopoietic stem cells (HSCs). IRF8 binds directly to an IRF8/Ets consensus sequence located in promoter regions of Sfpi1 and Ebf1, which encode PU.1 and EBF, respectively, and is associated with transcriptional repression of Sfpi1 and transcriptional activation of Ebf1. Bone marrows of IRF8 knockout mice (IRF8−/−) had significantly reduced numbers of pre-pro-B cells and increased numbers of myeloid cells. Although HSCs of IRF8−/− mice failed to differentiate to B220+ B-lineage cells in vitro, the defect could be rescued by transfecting HSCs with wild-type but not with a signaling-deficient IRF8 mutant. In contrast, overexpression of IRF8 in HSC-differentiated progenitor cells resulted in growth inhibition and apoptosis. We also found that IRF8 was expressed at higher levels in pre-pro-B cells than more mature B cells in wild-type mice. Together, these results indicate that IRF8 modulates lineage choice by HSCs and is part of the transcriptional network governing B-cell lineage specification, commitment, and differentiation.

Description: We report a detailed longitudinal study of the first patient to be treated (in 1973) for paroxysmal nocturnal hemoglobinuria (PNH) with syngeneic bone marrow transplantation (BMT). The patient subsequently relapsed with PNH in 1983, and still has PNH to date. Analysis of thePIG-A gene in a recent blood sample showed in exon 6 an insertion-duplication causing a frameshift. Polymerase chain reaction (PCR) amplification of the PIG-A exon 6 from bone marrow (BM) slides obtained before BMT showed that the duplication was not present; instead, we found several single base pair substitutions in exons 2 and 6. Thus, relapse of PNH in this patient was not due to persistence of the original clones; rather, it was associated with the emergence of a new clone. These findings support the notion that the BM environment may create selective conditions favoring the expansion of PNH clones. © 1998 by The American Society of Hematology.

Description: Heme oxygenase-1 (HO-1) is an intracellular enzyme that degrades heme and inhibits immune responses and inflammation in vivo. In most cell types, HO-1 is inducible by inflammatory stimuli and oxidative stress. Here we demonstrate that human monocyte-derived immature dendritic cells (iDCs) and several but not all freshly isolated rat splenic DC subsets and rat bone marrow-derived iDCs, spontaneously express HO-1. HO-1 expression drastically decreases during human and rat DC maturation induced in vitro. In human tissues, iDCs also express HO-1, whereas mature DCs do not. Induction of HO-1 expression with cobalt protoporphyrin (CoPP) in human and rat DCs inhibits lipopolysaccharide (LPS)-induced phenotypic maturation and secretion of proinflammatory cytokines, resulting in the inhibition of alloreactive T-cell proliferation. CoPP-treated DCs, however, retain the ability to produce the anti-inflammatory cytokine interleukin 10 (IL-10). Reactive oxygen species induced by LPS in DCs were inhibited by induction of HO-1. In conclusion, we identify, for the first time, the capacity of HO-1 to block maturation of DCs and to inhibit proinflammatory and allogeneic immune responses while preserving IL-10 production. This novel immune function for HO-1 may be of interest for the inhibition of immune responses in autoimmune diseases, transplantation, and other conditions involving activation of the immune system. (Blood. 2005;106:1694-1702)

Description: In order to establish an efficient gd T cell-mediated immunotherapy for hematological malignancies, we tried to clarify whether γδ T cells could be expanded from blood cells of patients with myeloma, lymphoma and acute leukemia by culture with zoledronate and a low dose of IL-2 and whether the expanded patients’ γδ T cells could kill tumor cells including self tumor cells with sparing normal clone cells. In addition, we explored the methods to enhance the anti-tumor cytotoxicity of the expanded γδ T cells by activating them with type I IFN, monocyte-derived dendritic cells (mo-DCs), or ab T cells. Although γδ T cells could be expanded in patients with myeloma, lymphoma and leukemia as well as normal persons, the amplification rates of gd T cells before and after the culture were varied from patient to patient in the patients with hematological malignancies. γδ T cells generated in patients with myeloma and lymphoma showed a potent cytotoxic ability against myeloma/lymphoma cell lines (RPMI8226, Daudi) as shown in γδ T cells generated in normal persons. In addition, γδ T cells generated in a patient with myeloma and acute leukemia showed a cytotoxic ability against self myeloma or leukemia cells freshly prepared from bone marrow. However, the same γδ T cells were not cytotoxic to normal lymphocytes of the patients. Then the expanded γδ T cells were stimulated with type I IFN, mo-DCs, or αβ T cells and the activation (CD69 expression) and cytotoxicity against tumor cells were examined. By the stimulation with type I IFN, the expression of CD69 and Trail of γδ T cells was increased and the cytotoxic ability of γδ T cells was enhanced at dose-dependent manner of type I IFN. CD69 expression on γδ T cells was enhanced by co-culture with both immature and mature mo-DCs in a cell-number-dependent fashion. CD69 expression was enhanced after the addition of mo-DCs of either autologous or allogeneic origin. Activation of γδ T cells with mo-DCs enhanced anti-tumor cytotoxicity of γδ T cells against RPMI8226 and CML blastic crisis cell line (C2F8) in an effector-to-target ratio-dependent manner. Although CD69 expression of γδ T cells was enhanced by the co-culture with allogeneic ab T cells, autologous ab T cells couldn’t activate γδ T cells. However, autologous ab T cells stimulated with IL-2 or PHA could induce the activation of γδ T cells. The activation of γδ T cells with stimulated αβ T cells required cell-to-cell interaction. These findings suggested that αβ T cells stimulated by allogeneic γδ T cells could activate the same allogeneic γδ T cells. The present data demonstrated that γδ T cells, which could be expanded in vitro from blood cells of the patients with myeloma, lymphoma and leukemia by culture with zoledronate and IL-2, possess an enough cytotoxic ability against tumor cells including self tumor cells with sparing normal cells. These findings suggested that in vitro generated patients’ γδ T cells could be applied to γδ T cell-mediated immunotherapy for hematological malignancies. Besides, potent γδ T cells activated by type I IFN, mo-DCs or activated αβ T cells were considered to be applicable for γδ T cell-mediated immunotherapy.

Description: Vascular endothelial growth factor (VEGF), which induces angiogenesis and increases vascular permeability, is a major growth factor mediating tumor progression. In this study, we employed immunohistochemical-staining method to detect the expression of VEGF in lymph nodes taken from39 non-Hodgkin’s lymphomas patients and analyzed the relation of the expression levels to malignant aggressiveness, treatment response, histological grade, clinical stage and prognosis. The patients had been observed for at least 5 years or until death. 9 patients with benign lymphadenopathy were acted as control. The expression of VEGF was assessed according to the percentage of immunoreactive cells in a total of 1000 neoplastic cells (quantitative analysis). Immunoreactivity was graded positive, more than 10% of carcinoma cells stained and negative, no detectable staining or less than 10% of carcinoma cells stained. Furthermore, the qualitative intensity of staining for VEGF was assessed using a scale of 0–3+. The expression analysis of VEGF revealed that in 31 out of 39 (79.49%) specimens VEGF staining was positive. The VEGF staining was always cell membrane. Significant associations were found between the expression of VEGF and histological grade, Ann Arbor stage, prognosis (according to International Prognostic Index, IPI) and chemotherapy response. Among 8 cases of low grade, 7 had lower-level expression and 1 had higher-level expression, but among 31 cases of intermediate and high grade, 13 had lower-level expression and18 had higher-level expression (P=0.044

Description: We examined a large cohort (N=2,457) of chronic lymphocytic leukemia (CLL) patients evaluated by the CLL Research Consortium (CRC) and found 63 (2.6%) used IGHV3-21. Comparing the Ig heavy chain third complementarity determining region (HCDR3) of the IGHV3-21 cases: 25/63 cases (39.7%) had a conserved amino acid motif (motif 1: DANGMDV) in the otherwise highly variable Ig HCDR3, as described by Tobin et al. Blood 2003. All but one of these Ig heavy chains (IgH) were paired with a lambda light chain encoded by IGLV3-21. In addition, we found that 3/63 cases (4.8%) had a previously unrecognized conserved HCDR3 amino acid motif (motif 2: DPSFYSSSWTLFDY). In contrast, these IgH invariably were paired with kappa immunoglobulin light chains (IgL) encoded by IGKV3-20. Similarly to that noted for CLL cases that use IgH encoded by unmutated IGHV1-69 (Widhopf et al. Blood Epub First Edition 2007), the pairing of IgH encoded by IGHV3-21 with IgL appears governed by the HCDR3. The non-stochastic pairing of IgH with IgL argues strongly that antigen plays a role in selecting the Ig expressed in CLL. To examine for the antigen(s) recognized by the most common Ig encoded by IGHV3-21, we isolated IgH and IgL genes expressed by IGHV3-21/IGLV3-21 CLL cases and generated recombinant antibodies, which we examined for binding to antigen(s) present on microarray of self or environmental antigens. We found that Ig encoded by IGHV3-21/IGLV3-21 had apparent specific binding for protein L, a multi-domain cell-wall protein isolated from Peptostreptococcus magnus, a Gram-positive commensal bacteria that comprise a large portion of the human bacterial gut flora. Prior studies identified that protein L is a superantigen capable of binding human Ig kappa light chains encoded by IGKV genes of the I, III, and IV subgroups, but not human Ig lambda light chains. The specific binding of IGHV3-21/IGLV3-21 to protein L suggested that protein L might play a role in the development of CLL cells that express such Ig. To test this hypothesis, we examined the capacity of various recombinant antibodies to bind protein L by ELISA. We found that lambda IgL encoded by IGLV3-21 could bind to protein L with similar activity, independent of whether this lambda IgL paired with the native IgH, IgH encoded by IGHV3-21 lacking the DANGMDV HCDR3 motif, or even irrelevant IgH encoded by IGHV4-39 that are not found paired with IGLV3-21 in the Ig expressed in CLL. Moreover, Ig formed by pairing IgH encoded by IGHV3-21 that has the DANGMDV HCDR3 motif with an IgL encoded by an IGLV that was irrelevant to IGLV3-21 did not bind protein L. These results reveal a previously unrecognized capacity of human IgL encoded by IGLV3-21 to bind the protein L superantigen of Peptostreptococcus magnus, a bacteria commonly found in the human gastrointestinal tract. However, because the binding of IGLV3-21 does not depend upon the non-stochaistic pairing of IgH and IgL observed in CLL, we reason that the capacity of IGLV3-21 to bind protein L cannot account for the selected Ig repertoire expressed in CLL, suggesting that it actually does not play a role in CLL leukemogenesis. This finding suggests that caution should be exercised when defining an antigen that is found capable of binding the restricted Ig expressed in CLL as the driving factor responsible for leukemogenesis.

Description: Background: Heavy chain disease (HCD) is a rare lymphoproliferative disorder characterized by a monoclonal heavy chain (HC) unattached to a light chain (LC). IgGHCD or γHCD typically presents as a lymphoproliferative disorder with lymphadenopathy and hepatosplenomegaly. Myeloma has been described associated with γHCD but only with a second intact Ig paraprotein. This report describes a unique presentation of multiple myeloma with monoclonal free γ3HC and kappa free light chains. Case: A 34 year old gentleman presented with mild persistent neutropenia following two episodes of pneumonia, 18 months previously. He admitted to persistent night sweats but no other significant history. Baseline investigations revealed a mild anaemia, neutropenia and a large IgG paraprotein with no associated light chain. Bone marrow aspirate and trephine confirmed myeloma. The patient was treated with cyclophosphamide, thalidomide and dexamethasone and has had a very good partial remission. He is awaiting a sibling allogeneic peripheral blood stem cell transplant. Investigations and results: Serum Electrophoresis confirmed a large IgG paraprotein (23g/l) with no associated light chain in the serum and identified as γ3 subclass by radial immunodiffusion. Western blot showed the γ3HC was truncated with a large deletion. Markedly elevated free kappa (κ) LC (503.58 mg/l [3.30–19.4]) were found in the serum with gross skewing of the kappa/lambda ratio. Urine electrophoresis revealed separate γHC and κ LC paraproteins. Western blot of the fractionated urine protein demonstrated different sized κLC aggregates. Flow cytometry of the marrow aspirate revealed an unusual staining pattern; CD5,19,38,45+ve and CD20,22,23,34,56,138 –ve plasma cells. Cytoplasmic staining revealed 2 distinct populations of plasma cells, the first producing γ3HC and the second only free κLC. Cytogenetics and FISH analysis for 14q, p53 and c-myc abnormalities were normal. Discussion: This is the first description of a Biclonal Myeloma with separate plasma cell populations producing γ3HC and κLC paraproteins. The biclonality confirms the free HC occurs as a result of abnormal synthesis not cleavage. The clinical and immunological findings are clearly different to typical findings in both γ3HCD and Myeloma. HCD has an appalling prognosis and this case is likely to have been ‘smouldering’ for 18 months, evidenced by the 2 pneumonias and persistent night sweats. There is no lymphadenopathy or organomegaly associated with γ3HCD. The immunophenotype of the malignant plasma cells is unique. Other atypical features include frank proteinuria, with a HC in the urine, but normal renal function and no radiological or biochemical evidence of bone involvement. We propose that this unique biclonal myeloma has distinct immunological and clinical features.

Description: To realize the therapeutic potential of human embryonic stem cells (hESCs), it is necessary to regulate their differentiation in a uniform and reproducible manner. We have developed a method in which known numbers of hESCs in serum-free medium were aggregated by centrifugation to foster the formation of embryoid bodies (EBs) of uniform size (spin EBs). These spin EBs differentiated efficiently and synchronously, as evidenced by the sequential expression of molecular markers representing stem cells, primitive streak, and mesoderm. In the presence of hematopoietic growth factors, reproducible differentiation was achieved with blood cells formed in more than 90% of EBs. Using chimeric EBs generated from mixtures of green fluorescence protein–positive (GFP+) and GFP– hESCs in a clonogenic assay, hematopoietic precursor frequency was estimated to be approximately 1:500 input cells. This method of EB formation provides a generally applicable means for modulating and objectively monitoring the directed differentiation of hESCs.

Description: Morphological and functional modifications occurring in Langendorff rabbit heart preparations perfused with purified human leukocytes (PMNL), as an organ model of sulfidopeptide-leukotrienes (sLT) transcellular biosynthesis, were studied. Coronary perfusion pressure (CPP), monitored as an index of coronary vasospasm, increased by 295% after challenge with the Ca(2+)-ionophore A-23187 (0.5 micromol/L) for 30′, accompanied by a significant formation of sLT. Increase in CPP was prevented by PMNL pretreatment with the 5-lipoxygenase inhibitor MK-886 (1 micromol/L) or by heart pretreatment with LTD4-receptor antagonist SKF 104353, indicating a pivotal role of PMNL-derived 5-lipoxygenase (5- LO) products in the observed functional modifications. Similar effects were obtained using granulocyte macrophage-colony stimulating factor- primed PMNL challenged with the tripeptide n-formyl-methionyl-leucyl- phenylalanine. Scanning electron microscopy (SEM) of coronary arteries showed craters on the vessel luminal surface, PMNL adhering to endothelial cells (EC), increased number of microvilli on EC, presence of nonviable, desquamating, fusiform EC. SEM and transmission electron microscopy of myocardial microvessels, showed presence of perivascular and intermuscle edema, presence of activated PMNL and decreased number of patent microvessels. These morphological alterations were significantly blunted by MK-886 or SKF 104353. These data provide evidence of close interaction between PMNL and myocardial EC, resulting in enhanced sLT formation via transcellular biosynthesis, originating from transfer of PMNL-derived LTA4 to EC. These potent proinflammatory autacoids are responsible for coronary vasospasm and the morphological alternations observed.

Description: A distinct feature of multiple myeloma (MM) is the tight interaction between malignant plasma cells and their bone microenvironment, creating a niche suitable for MM growth. In particular, MM cells inhibit osteoblast (OB) differentiation and stimulate osteoclast (OC) function, resulting in imbalanced bone remodeling and osteolytic bone disease. Here we studied a novel cytokine, activin A, identified from a broad range of cytokines, in the development of MM bone disease. We next asked whether activin A inhibition could restore bone balance and suppress tumor growth. Activin, a member of the TNF-α superfamily, is a pleiotropic cytokine involved in bone remodeling. Here, we observed, that MM patients with multiple osteolytic lesions had a 4-fold increase in activin A expression levels in bone marrow plasma compared to MM patients with one or less osteolytic lesions and non-MM patients (average 123.6 ± 136 vs 26.4 ± 21.4 vs 30.6 ± 25.1 pg/ml respectively, p

Description: Gene transfer into human hematopoietic stem cells with expression targeted to the maturing myelomonocytic progeny has applications for gene therapy of genetic diseases affecting granulocytes and macrophages. We hypothesized that promoters of myeloid-specific genes that are upregulated with myelomonocytic differentiation would also upregulate expression of an exogenous gene in a retroviral vector. Moloney murine leukemia virus (MoMuLV)-based retroviral vectors using promoters from hematopoietic genes (CD11b, CD18, and CD34) were compared with vectors with viral promoters (MoMuLV long terminal repeat [LTR], cytomegalovirus [CMV], and simian virus 40 [SV40]). Human glucocerebrosidase (GC) cDNA was the reporter gene. HL60 cells were transduced with these vectors and vector-derived GC activity was compared in undifferentiated HL-60 cells and the same cells differentiated into granulocytes using dimethyl sulfoxide or monocyte/macrophages using phorbol myristate acetate. In undifferentiated HL-60 cells, vector-derived GC activity was the highest when it was controlled by the MoMuLV LTR. In HL-60 cells differentiated into granulocytes, vector-derived GC activity transcribed from the CD11b, MoMuLV LTR, and CMV promoters was equivalent to 1.7, 1.5, and 1.5 times the normal endogenous GC activity, respectively, and 0.8, 2.0, and 3.6 times the normal GC activity, respectively, in those differentiated into macrophages. With granulocytic differentiation, the CD11b promoter showed maximal induction in GC activity (8-fold); with macrophage differentiation, the CD11b promoter showed a fourfold induction in GC expression. The CD11b promoter also generated significant levels of GC activity in the myelomonocytic progeny of transduced CD34+ cells. Expression from the CD11b promoter, unlike that from the CMV or the MoMuLV LTR promoters, was relatively myelomonocyte-specific, with minimal expression observed in Jurkat T cells or HeLa carcinoma cells. The induction of expression from the CD11b promoter with differentiation in HL-60 cells correlates with the developmental regulation of the CD11b gene. Retroviral vectors using the CD11b promoter have potential utility for gene therapy of disorders affecting the myelomonocytic lineage.

Description: The past decade has shown a marked increase in the use of high-throughput assays in clinical research into human cancer, including acute myeloid leukemia (AML). In particular, genome-wide gene expression profiling (GEP) using DNA microarrays has been extensively used for improved understanding of the diagnosis, prognosis, and pathobiology of this heterogeneous disease. This review discusses the progress that has been made, places the technologic limitations in perspective, and highlights promising future avenues

Description: As previously reported (Colombat, Blood2001;97:101), rituximab (4 weekly doses of 375mg/m²) can lead to high response rates (RR) and prolonged remissions with minimal toxicity as 1st line therapy for low tumor burden FL. We report the final analysis of a trial evaluating long term efficacy and safety of rituximab in untreated low tumor burden FL (GELF criteria). 49 patients (pts) were included in the initial trial (median age 52 yrs), 2 refused consent for the extended F/Up period, and 1 pt died at M12. Molecular bcl2-JH rearrangement was assessed throughout the study. The median F/up was 83.8 mths. Overall best RR, complete/unconfirmed RR and partial RR at D78 were 74%, 50% and 24% respectively. Median PFS was 23.5 mths for the study population. Median duration of response (34 responders at D78, i.e 6 weeks after the last rituximab dose) was 28.6 mths, but response was still maintained without any further treatment in 11 pts after 5 years (24%) and in 7 pts after 7 years (15%). 31/46 pts were bcl2 positive in blood and/or marrow samples before rituximab: 11 (35%) became negative at D50, and 20 remained positive (65%). Median PFS was 37 mths for bcl2-negative pts at D50, and 12 mths for patients remaining positive (p=0.018 Log-rank). Of the 7 pts with sustained response after 7 years, 5 were bcl2 positive at D0, 2/5 became negative at D50, and 5/5 were still negative at M84. At year 7, 4/46 pts have died (1 from myelodysplasia, 3 from NHL), 35/42 have progressed, and 7 have never progressed without any other treatment than the initial rituximab therapy. Time to progression was significantly longer in the bcl2-negative population at D50 (p= 0.018, Log-rank). Duration of response was not correlated with bcl2 status at D50, but was associated with ‘Best response CR/Cru’ (p=0.007 Log-rank). Long-term tolerance was good, with only 13 SAE observed in 13 pts during the additional 4 years of F/Up (4 surgeries for non NHL-related pathologies, 1 node biopsy, 1 sleep apnea syndrome, 1 ischemic cardiopathy, 2 deaths from NHL, 1 depression, 1 pneumonia, 1 erysipela, 1 bronchitis). This long-term update confirms that a single 4-dose rituximab treatment yields durable benefits without the toxicity of chemotherapy for pts with low burden FL : Median PFS of 23.5 mths for the cohort, 28.6 mths for responders and 37 mths for pts turning bcl2-negative at D50, 15% of pts have maintained their response after 7 years, (2bis) the quality (CR/Cru) of the initial response was associated with a longer response duration high overall survival is observed with 4 deaths/46 pts (8.6%).

Description: Background: Recent advances in the therapy of multiple myeloma (MM) have greatly increased the treatment options for this uniformly fatal plasma cell malignancy. It is not clear if introduction of novel therapies and the increased use of high dose therapy (HDT) in the past decade have translated into better outcome for patients (pts) with MM. Methods: We examined the outcome of two cohorts of pts seen at our institution. The first cohort consisted of 387 pts who relapsed after HDT and was examined for potential improvement in survival following relapse after HDT. These pts were divided into two groups; those who relapsed before or after December 31, 2000. The second cohort consisted of 2981 patients with newly diagnosed MM seen between January 1971 and December 2006 and was used to examine the trends in overall survival (OS). Results: Among those relapsing after HDT, there were 245 males (63%); median (range) age at HDT was 57 years (32.8–75.4) and the median time to HDT was 8.1 months (1–90 months) from diagnosis. The median time to relapse was 13.2 months (1.1 months-10.3 years) from HDT. In this cohort, a clear improvement in OS from time of relapse was seen over the past decade, with those relapsing after 2000 having a median OS of 23.9 months (95% CI; 19.8, 27.6) compared to 11.8 months (95% CI; 8.7, 14.9) for the rest (P 〈 0.001) (Figure 1). In a multivariate analysis, the effect of the date of relapse on survival was independent of other prognostic factors such as relapse 5.5 mg/dL. Pts who were treated with one or more of the newer drugs (thalidomide, lenalidomide, bortezomib) had longer survival from relapse (30.9 months (95% CI; 23.6, 38.2) compared to 14.8 months (95% CI; 11.3, 18.4); P 〈 0.001) for others. Among the newly diagnosed MM cohort, the median age at diagnosis was 66 years (20.2 – 97 years), and 1,770 (59.4%) were males. The median follow up for the entire group was 27.4 months (0–29.4 years); and at the time of analysis 558 patients (18.7%) were alive with a median follow up of 32.7 months (0–29.4 years). Pts diagnosed in the last decade had an improved OS (44.8 months) compared to those diagnosed before this period (29.9 months; P 〈 0.001) (Figure 2). The improvement in survival seen in the last decade among newly diagnosed patients was predominantly among those younger than 65 years (60.3 mos vs. 33.3 mos improvement in median survival); compared to those over 65 at diagnosis (26.5 mos vs. 32 mos). Conclusion: In this study, for the first time, we demonstrate definite proof for improved outcome in patients with myeloma, both in the relapsed setting as well as at diagnosis, a change that is likely to continue with increased use of these drugs. Figure Figure Figure Figure

Description: INTRODUCTION: Proteasome inhibitors (PI) and histone deacetylase inhibitors (HDACi) have demonstrated synergistic pre-clinical activity in multiple myeloma (MM). The goals of this study were to evaluate this combination regimen’s clinical activity and adverse events, including thrombocytopenia (TCP) since both drug classes may cause transient TCP. We performed an open label, single-centre, single-arm, phase I/II, dose-escalation trial of bortezomib, dexamethasone and romidepsin (depsipeptide) in relapsed or refractory MM. This is the first clinical trial to combine these 3 agents. METHODS. All patients (pts) received bortezomib (1.3mg/m2 d1, 4, 8, 11) with dexamethasone (20mg d1, 2, 4, 5, 8, 9, 11, 12). Romidepsin commenced at 8 mg/m2 IV d1, 8, and 15 every 28 days with a planned accelerated intra-patient dose escalation to 10, 12 and 14 mg/m2 (n=10). After CR + 2 cycles or a maximum of 8 cycles, pts with SD or better commenced maintenance (Mx) therapy, romidepsin at the MTD on days 1 and 8 of a 28 day cycle until PD. An additional 15 pts were treated at the MTD in a phase II expansion. Response was assessed after every 2 cycles according to IMWG criteria (with minimal Response (MR) defined as ≥25% but

Description: Endothelial cells secrete prothrombotic ultralarge von Willebrand factor (VWF) multimers, and the metalloprotease ADAMTS13 cleaves them into smaller, less dangerous multimers. This reaction is stimulated by tensile force applied to the VWF substrate, which may occur on cell surfaces or in the circulating blood. The cleavage of soluble VWF by ADAMTS13 was accelerated dramatically by a combination of platelets and fluid shear stress applied in a cone-plate viscometer. Platelet-dependent cleavage of VWF was blocked by an anti-GPIbα monoclonal antibody or by a recombinant soluble fragment of GPIbα that prevents platelet-VWF binding. Multimeric gel analysis showed that shear and platelet-dependent cleavage consumed large VWF multimers. Therefore, ADAMTS13 preferentially acts on platelet-VWF complexes under fluid shear stress. This reaction is likely to account for a majority of VWF proteolysis after secretion and to determine the steady-state size distribution of circulating VWF multimers in vivo.

Description: Abstract 1689 Poster Board I-715 Introduction The use of the proteasome inhibitor bortezomib has demonstrated activity in multiple myeloma and lymphomas. The HDAC inhibitor romidepsin is being evaluated in CTCL and PTCL, though its activity in B-cell lymphomas is less clear. We hypothesized that the combination of bortezomib and romidepsin would result in synergistic apoptosis in different B-cell NHL cell lines based upon the observed activity of this combination in more mature B-cell malignancies such as myeloma. Experimental Design Daudi, HT, Ramos and SUDHL-4 cell lines were exposed to different concentrations of bortezomib and romidepsin, separately, concurrently, and sequentially. Cell viability was assessed using MTT-assay, induced apoptosis was evaluated using Annexin V and PI staining from 24-48 hours. Apoptosis was also evaluated using western blot analysis of caspases and PARP cleavage. LC3 and HDAC6 level expressions were performed to determine if the effect of the combination was a result of the aggresome or autophagy pathway. Cell cycle studies were also performed to study if there were any changes after treating cells with the combination. Results The combination of bortezomib and romidepsin resulted in synergistic B-cell apoptosis as measured by MTT-assay with combination indices of 〈 0.5. This was associated with increased caspases and PARP cleavage as early as 24 hours after exposure. Order of addition experiments demonstrated definite sequence specificity. When romidepsin was added first, and 6 hours later followed by bortezomib, apoptosis was enhanced, compared to both agents being given concurrently or when bortezomib was administered first. Cell cycle analysis studies demonstrated that pretreatment of cells with romidepsin for 6 hours followed by the addition of bortezomib arrested the cells in G2M phase. HDAC6 expression was significantly reduced following combination therapy, and LC3-I was cleaved to LC3-II in treated cells suggesting that the combination affected aggresome formation and autophagy. Conclusion The combination of romidepsin and bortezomib at low nanomolar concentrations suggests that this may be an important clinical combination to test in patients with relapsed or refractory B-cell malignancies. Sequence of administration data is currently being tested to determine if the effect is a result of autophagy inhibition as is seen in myeloma cell lines. Additional mechanistic studies will be presented with the goals of identifying predictors of response that can then be validated in prospective clinical trials. Disclosures Lechowicz: Gloucester: Consultancy. Kaufman:Millennium: Consultancy; Genzyme: Consultancy; Celgene: Consultancy; Merck: Research Funding; Celgene: Research Funding. Lonial:Gloucester: Research Funding; Novartis: Consultancy; BMS: Consultancy; Millennium: Consultancy, Research Funding; Celgene: Consultancy. Flowers:Millennium: Research Funding.

Description: Abstract 3701 Poster Board III-637 Background Romidepsin is an anti-neoplastic agent that has been identified as a novel pan-HDAC inhibitor with single-agent activity in T-cell lymphoma. In a combined analysis of 167 patients (pts) with cutaneous T-cell lymphoma (CTCL) from 2 clinical studies (GPI and NCI studies), the overall response rate was 35%, including 10 pts with a complete clinical response (CCR). Median duration of response was 13.8 months and 42% of pts with advanced disease (stage ≥IIB) responded [Demierre et al. J Clin Oncol 27:15s, 2009 (suppl; abstr 8546)]. The most common hematologic abnormalities in these pts included anemia (41%), thrombocytopenia (34%), neutropenia (27%), and lymphopenia (26%). Most hematologic toxicities were Grade 1 or 2, although 'Grade 3 events were observed. These events were reversible and a small portion of the patients discontinued the study drug because of these events (2%). This report details an analysis of platelet counts in pts receiving romidepsin and an investigation into the mechanism of thrombocytopenia in nonclinical studies. Methods Pts with CTCL who received ≥1 prior systemic therapy failure and had an ECOG PS of 0-2 were enrolled in 2 single-arm, open-label, multicenter and international clinical studies. Treatment with QTc prolonging therapies or CYP34A inhibitors was prohibited and pts with significant cardiovascular abnormalities were excluded. Romidepsin at 14 mg/m2 was administered as a 4-hr IV infusion on days 1, 8, and 15 of a 28 day cycle. Nonclinical studies were conducted in mice to investigate the mechanism of romidepsin effects on platelets. Romidepsin was administered to female BALB/c mice at doses of 1 or 4 mg/kg by tail-vein injection on days 1, 5 and 9. Blood samples were collected every 2 days from alternating groups of mice to minimize effects of bleeding on platelet counts. Results In clinical studies, there is a mean decrease in platelet counts during the treatment period of each cycle, and a return to baseline levels or above between cycles observed in both clinical studies as described in the table below. No clinically meaningful change has been observed in the central tendency over 4 cycles of treatment in both studies. In the mouse studies, dose-dependent effects were seen on both WBC and platelet counts. Day 2 WBC counts dropped to 45% and 10% of normal at the 1 and 4 mg/kg doses, respectively. WBC counts remained low until after the dosing period in the 1 mg/kg romidepsin group, but recovered more quickly in the 4 mg/kg group. Day 2 platelet counts were 70% of normal at the 1 mg/kg dose and remained near this level until day 10, followed by recovery to normal at day 15. At the 4 mg/kg dose, profound thrombycytopenia was induced, with platelet counts only 20% of normal on days 4-6. Platelet counts slowly recovered to 70% of normal by day 15. Plasma thrombopoietin levels were normal throughout the experiment for the 1 mg/kg group, and showed a large increase to 275% of normal on day 6 in the 4 mg/kg group, which is the expected response to thrombocytopenia as a signal to increase platelet production and indicates that platelet reduction is not attributable to defective TPO production. Bone marrow megakaryocyte populations are being examined to determine the effects of romidepsin on these platelet-producing cells. Conclusions Following romidepsin administration, a saw tooth pattern is observed in the reduction and recovery of platelets. Recovery of platelets appears to occur more rapidly in humans than in mice; however, the effects are reversible after dosing in clinical studies and in murine models. In the clinical data the recovery pattern suggests that the transient effects are direct and are not effects on bone marrow. Disclosures: Whittaker: Gloucester Pharmaceuticals: Research Funding. Prince:Gloucester Pharmaceuticals: Consultancy. Demierre:Gloucester Pharmaceuticals: Consultancy, Honoraria. Lonial:Gloucester Pharmaceuticals: Honoraria. Kim:Gloucester Pharmaceuticals: Consultancy, Honoraria. Nichols:Gloucester Pharmaceuticals: Employment, Equity Ownership. Nix:Gloucester Pharmaceuticals: Employment.

Description: Precursor T-cell acute lymphoblastic leukemia (T-ALL) in children represents a clinical challenge, because relapses are usually fatal. It is thus necessary to identify high-risk patients as early as possible to effectively individualize treatment. We aimed to define novel molecular risk markers in T-ALL and performed array-based comparative genomic hybridization (array-CGH) and expression analyses in 73 patients. We show that DNA copy-number changes are common in T-ALL and affect 70 of 73 (96%) patients. Notably, genomic imbalances predicted to down-regulate the TGF-β or up-regulate the PI3K-AKT pathways are identified in 25 of 73 (34%) and 21 of 73 (29%) patients, suggesting that these pathways play key roles in T-ALL leukemogenesis. Furthermore, we identified a deletion at 6q15-16.1 in 9 of 73 (12%) of the patients, which predicts poor early treatment response. This deletion includes the CASP8AP2 gene, whose expression is shown to be down-regulated. The interaction of CASP8AP2 with CASP8 plays a crucial role in apoptotic regulation, suggesting a functional link between the clinical effect of the deletion and the molecular mode of action. The data presented here implicate the TGF-β and PI3K-AKT pathways in T-ALL leukemogenesis and identify a subgroup of patients with CASP8AP2 deletions and poor early treatment response.

Description: Absence of shear stress due to disturbed blood flow at arterial bifurcations and curvatures leads to endothelial dysfunction and proinflammatory gene expression, ultimately resulting in atherogenesis. KLF2 has recently been implicated as a transcription factor involved in mediating the anti-inflammatory effects of flow. We investigated the effect of shear on basal and TNF-α–induced genomewide expression profiles of human umbilical vein endothelial cells (HUVECs). Cluster analysis confirmed that shear stress induces expression of protective genes including KLF2, eNOS, and thrombomodulin, whereas basal expression of TNF-α–responsive genes was moderately decreased. Promoter analysis of these genes showed enrichment of binding sites for ATF transcription factors, whereas TNF-α–induced gene expression was mostly NF-κB dependent. Furthermore, human endothelial cells overlying atherosclerotic plaques had increased amounts of phosphorylated nuclear ATF2 compared with endothelium at unaffected sites. In HUVECs, a dramatic reduction of nuclear binding activity of ATF2 was observed under shear and appeared to be KLF2 dependent. Reduction of ATF2 with siRNA potently suppressed basal proinflammatory gene expression under no-flow conditions. In conclusion, we demonstrate that shear stress and KLF2 inhibit nuclear activity of ATF2, providing a potential mechanism by which endothelial cells exposed to laminar flow are protected from basal proinflammatory, atherogenic gene expression.

Description: The cAMP-responsive element binding protein (CREB) is a 43-kDa nuclear transcription factor that regulates cell growth, memory, and glucose homeostasis. We showed previously that CREB is amplified in myeloid leukemia blasts and expressed at higher levels in leukemia stem cells from patients with myeloid leukemia. CREB transgenic mice develop myeloproliferative disease after 1 year, but not leukemia, suggesting that CREB contributes to but is not sufficient for leukemogenesis. Here, we show that CREB is most highly expressed in lineage negative hematopoietic stem cells (HSCs). To understand the role of CREB in hematopoietic progenitors and leukemia cells, we examined the effects of RNA interference (RNAi) to knock down CREB expression in vitro and in vivo. Transduction of primary HSCs or myeloid leukemia cells with lentiviral CREB shRNAs resulted in decreased proliferation of stem cells, cell- cycle abnormalities, and inhibition of CREB transcription. Mice that received transplants of bone marrow transduced with CREB shRNA had decreased committed progenitors compared with control mice. Mice injected with Ba/F3 cells expressing either Bcr-Abl wild-type or T315I mutation with CREB shRNA had delayed leukemic infiltration by bioluminescence imaging and prolonged median survival. Our results suggest that CREB is critical for normal myelopoiesis and leukemia cell proliferation.

Description: Graft-versus-host disease (GVHD) is currently encountered after bone marrow transplantation and transfusion. GVHD associated with transfusion (TA-GVHD) in apparently immunocompetent recipients has been recently reported with increasing frequency. A consistent finding in many of these cases is that the recipient received blood from a donor homozygous for one of the recipient's HLA haplotypes. However, the observed frequency of TA-GVHD is much lower than the estimated probability of this donor/recipient combination. The potential role of recipient immune responses in controlling TA-GVHD was investigated using an analogous murine model in which GVHD is induced by the injection of parental lymphoid cells into unirradiated F1 hybrid recipients. The effect of various immune manipulations of the recipient of GVHD induction was assessed by determining the number of donor lymphoid cells required to induce GVHD responses. Whereas depletion of recipient CD4+ cells increased the number of donor cells needed to induce GVHD, depletion of recipient CD8+ and natural killer cells resulted in fewer donor cells being needed to induce a GVHD response. These studies suggest a central role for functioning recipient CD8 and natural killer cells in the down-regulation of TA-GVHD development in recipients.

Description: Bone marrow transplantation (BMT) is relatively effective for the treatment of lysosomal storage diseases. To better understand the contribution of specific hematopoietic lineages to the efficacy of BMT, we transplanted β-glucuronidase–positive mononuclear phagocytes derived from either the peritoneum or from bone marrow in vitro into syngeneic recipients with mucopolysaccharidosis type VII (MPS VII). Cell surface marking studies indicate that the bone marrow-derived cells are less mature than the peritoneal macrophages. However, both cell types retain the ability to home to tissues rich in cells of the reticuloendothelial system after intravenous injection into MPS VII mice. The half-life of both types of donor macrophages is approximately 7 days, and some cells persist for at least 30 days. In several tissues, therapeutic levels of β-glucuronidase are present, and histopathologic analysis demonstrates that lysosomal storage is dramatically reduced in the liver and spleen. Macrophages intravenously injected into newborn MPS VII mice localize to the same tissues as adult mice but are also observed in the meninges and parenchyma of the brain. These data suggest that macrophages play a significant role in the therapeutic efficacy of BMT for lysosomal storage diseases and may have implications for treatments such as gene therapy.

Description: In anaplastic large-cell lymphoma (ALCL), the (2;5) chromosomal translocation creates a fusion gene encoding the 80-kD NPM-ALK hybrid protein. This report describes three new monoclonal antibodies, two of which recognize, by Western blotting, the N-terminal portion of NPM present in the NPM-ALK fusion protein and also in two other NPM fusion proteins (NPM-RAR and NPM-MLF1). The third antibody recognizes the C-terminal portion (deleted in NPM-ALK) and reacts only with wild-type NPM. The three antibodies immunostain wild-type NPM (in paraffin-embedded normal tissue samples) in cell nuclei and in the cytoplasm of mitotic cells. Cerebral neurones, exceptionally, show diffuse cytoplasmic labeling. In contrast to normal tissues, the two antibodies against the N-terminal portion of NPM labeled the cytoplasm of neoplastic cells, in four ALK-positive ALCL, reflecting their reactivity with NPM-ALK fusion protein, whereas the antibody to the C-terminal NPM epitope labeled only cell nuclei. Immunocytochemical labeling with these antibodies can therefore confirm that an ALK-positive lymphoma expresses NPM-ALK (rather than a variant ALK-fusion protein) and may also provide evidence for chromosomal anomalies involving the NPM gene other than the classical (2;5) translocation.

Description: Venous thromboembolism (VTE) is increasingly diagnosed among individuals with hematologic malignancies. However, the risk of VTE among patients undergoing hematopoietic stem cell transplantation (HSCT) is unclear. We examined the incidence and risk factors for VTE and bleeding among 1514 patients undergoing in-patient HSCT. No protocolized VTE prophylaxis was used. By HSCT day 180, 75 symptomatic VTE occurred in 70 patients (4.6%; 95% confidence interval [CI], 3.6%-5.8%). Fifty-five (3.6%) were catheter-associated, 11 (0.7%) were non–catheter-associated deep venous thromboses, and 9 (0.6%) were pulmonary emboli. Thirty-four percent of VTE occurred at a platelet count less than 50 ×109/L; 13% occurred at a platelet count less than 20 ×109/L. In multivariate analysis, VTE was associated with prior VTE (odds ratio [OR], 2.9; 95% CI, 1.3-6.6) and with graft-versus-host disease (GVHD; OR, 2.4; 95% CI, 1.4-4.0). Clinically significant bleeding occurred in 230 patients (15.2%; 95% CI, 13.4%-17.1%); 55 patients (3.6%; 95% CI, 2.7%-4.7%) had fatal bleeding. Bleeding was associated with anticoagulation (OR, 3.1; 95% CI, 1.8-5.5), GVHD (OR, 2.4; 95% CI, 1.8-3.3), and veno-occlusive disease (OR, 2.2; 95% CI, 1.4-3.6). In HSCT patients, VTE is primarily catheter-related and 3-fold less common than clinically significant bleeding. These findings warrant consideration when selecting VTE prophylaxis in HSCT patients.

Description: Tumor cell–associated tissue factor (TF) is a powerful determinant of metastatic potential. TF may increase metastasis by supporting thrombin-mediated proteolysis, through intracellular signaling events mediated by the TF cytoplasmic domain, through TF/fVIIa/fXa–mediated activation of protease-activated receptors, or through a combination of these processes. To better define the relationship between tumor cell-associated TF and circulating hemostatic factors in malignancy, we generated a set of C57Bl/6-derived tumor lines genetically lacking TF, expressing wild-type murine TF, or expressing a mutant TF lacking the cytoplasmic domain. Comparison of the metastatic potential of these cells in immunocompetent mice with genetic deficits in prothrombin, platelet function, or fibrinogen revealed that TF supports metastasis through mechanisms independent of the cytoplasmic domain, but dependent on each of these distal hemostatic factors. TF was neither required for primary tumor growth nor necessary for initial localization of embolized tumor cells within the lungs. Rather, tumor cell fate studies indicated TF supports metastasis by increasing the survival of micrometastases. One mechanism linking TF to metastasis is through a fibrin(ogen)-dependent and platelet-dependent restriction in natural killer cell–mediated clearance of micrometastases. However, TF also supported the early success of micrometastases through an additional mechanism independent of natural killer cells, but coupled to circulating prothrombin.

Description: We describe a dominantly inherited β-thalassemia intermedia phenotype observed in a five-generation Portuguese family. Carriers are characterized by moderate anemia, hypochromia, microcytosis, elevated hemoglobin (Hb)A2 and HbF levels, splenomegaly, hepatomegaly, and inclusion bodies in pheripheral red blood cells after splenectomy. The molecular basis of this condition is a small deletion within the 5′ consensus splicing sequence of the second intron of the β-globin gene, IVS-II-4,5 (-AG). Reticulocyte RNA studies performed by reverse transcription-polymerase chain reaction (RT-PCR) and primer extension analysis showed three abnormally processed transcripts, which, upon sequencing, were shown to correspond to (1) skipping of exon 2, and (2) activation of two cryptic splice sites (between codons 59/60, and at IVS-II-47). In vitro translation studies of these patients' reticulocyte RNA have shown that at least one of these aberrant mRNA species is translated into an abnormally elongated peptide whose cytotoxic properties could, in part, be causing the atypical dominant mode of inheritance observed in this family. We suggest that this elongated β chain is unable to combine with an α-globin chain to form a functional Hb molecule. Its degradation would, then, exhaust the proteolytic defense mechanism of the erythroid precursors, leading to inefficient proteolysis of the free α chains in excess.

Description: Recent studies have demonstrated that in peripheral lymphoid tissues of normal mice and healthy humans, 1% to 5% of αβ T-cell receptor–positive (TCR+) T cells are CD4−CD8− (double-negative [DN]) T cells, capable of down-regulating immune responses. However, the origin and developmental pathway of DN T cells is still not clear. In this study, by monitoring CD4 expression during T-cell proliferation and differentiation, we identified a new differentiation pathway for the conversion of CD4+ T cells to DN regulatory T cells. We showed that the converted DN T cells retained a stable phenotype after restimulation and that furthermore, the disappearance of cell-surface CD4 molecules on converted DN T cells was a result of CD4 gene silencing. The converted DN T cells were resistant to activation-induced cell death (AICD) and expressed a unique set of cell-surface markers and gene profiles. These cells were highly potent in suppressing alloimmune responses both in vitro and in vivo in an antigen-specific manner. Perforin was highly expressed by the converted DN regulatory T cells and played a role in DN T-cell–mediated suppression. Our findings thus identify a new differentiation pathway for DN regulatory T cells and uncover a new intrinsic homeostatic mechanism that regulates the magnitude of immune responses. This pathway provides a novel, cell-based, therapeutic approach for preventing allograft rejection.

Description: When adopting basic principles learned in mice to clinical application in humans, it is often difficult to distinguish whether a “translation” fails because of an invalid target in the human disease or because the therapeutic agents are not optimal for the human target. It is, therefore, desirable to develop preclinical models to optimize therapies for human targets using in vivo settings. Although anti–mouse CTLA-4 antibodies are known to enhance immune responses in vivo, their effect on T-cell activation in vitro ranges from enhancement to inhibition. Here we use the hu-PBL-SCID mouse model of Epstein-Barr virus (EBV)–associated lymphoma development to screen a panel of anti–human CTLA-4 monoclonal antibodies (mAbs) for their effect on human lymphocytes in an in vivo “humanized” environment. We report significant heterogeneity of anti–human CTLA-4 mAbs in enhancing the expansion of human T cells in mice, and this heterogeneity cannot be attributed to immunoglobulin isotypes or affinity for CTLA-4. These data validate the development of additional screening tools, such as the one described, to further characterize functional activity of antihuman antibodies before proceeding with clinical translation to human studies.

Description: Abstract 1955 Poster Board I-978 The detection of chromosome abnormalities in mature B-cell neoplasms by conventional cytogenetics remains difficult, mainly due to the low proliferative rate of mature lymphoid cells. The current FISH panel for chronic lymphocytic leukemia (CLL) is designed to detect some of the more common abnormalities of prognostic significance in CLL [i.e., del(6q), del(11q), +12, del(13q), del(17p)]. This CLL FISH panel has improved the detection rate of these markers by making it possible to obtain cytogenetic information from interphase cells; however, as it is limited to only these 5 markers, it cannot detect all abnormalities associated with CLL. More importantly, the impact of other chromosome abnormalities on prognosis and disease progression, with and without the presence of these 5 prognostic markers, is not known. CpG-oligodeoxynucleotides (ODNs) such as DSP30 activate cells of the immune system in a sequence-dependent manner and promote proliferation of CLL cells in vitro [Decker et al. Blood 2000;95:999-1006]. They also upregulate costimulatory molecules and potential target antigens during immunotherapy. The use of DSP30 in combination with interleukin 2 (IL2) has proven effective in increasing the detection of chromosome abnormalities in CLL [Dicker et al. Blood 2006;108:3152-60] and other mature B-cell lymphoid malignancies by conventional cytogenetics [Struski et al. Leukemia 2009;23:617-9], when compared to the well-established B-cell mitogens. In our extensive clinical experience of incorporating DSP30/IL2 into our culture media, this cocktail has significantly increased the detection of chromosome abnormalities in CLL by conventional cytogenetics, from 55% to greater than 80%. We thus decided to investigate if various other lymphoid malignancies would respond to the mitogen activity of DSP30/IL2 as well as or better than CLL. Specifically, we evaluated 812 cases of mature B-cell lymphoid malignancies that were abnormal by flow cytometry, morphology, or cytogenetic analysis. All samples (bone marrow or blood) were cultured for approximately 72 hours using the DSP30/IL2 mitogen cocktail. Of these 812 cases, 746 (91%) provided sufficient mitotic index and quality for a complete cytogenetic analysis and interpretation. Of the CLL cases (n=509), 79 were initially interpreted as normal by conventional cytogenetic analysis, but were later interpreted as abnormal by FISH for deletion 13q only. In view of the known cryptic nature of this deletion in CLL, these cases were not included in the study, leaving a total of 430 CLL cases, and thus bringing the total number of cytogenetically successful study cases to 667. In addition to the 430 CLL cases, there were 14 variant CLLs; 36 diffuse large B-cell lymphomas (DLBCLs); 35 follicular lymphomas; 34 non-Hodgkin lymphomas (not further specified); 29 marginal zone B-cell lymphomas of splenic type (sMZBCL); 27 mantle cell lymphomas (MCLs), of which 8 were blastoid; 16 MZBCL of MALT type; 13 hairy cell leukemias (HCLs); 12 lymphoproliferative disorders (not further specified); 10 lymphoplasmacytic lymphomas (LPLs); 6 Burkitt lymphomas; 3 Hodgkin lymphomas; and 2 B-cell prolymphocytic leukemias (PLLs). Of particular interest is the fact that we detected clonal abnormalities in 100% of HCLs, blastoid MCLs, variant CLLs, and B-cell PLL, as well as in 97% of sMZBCLs, 89% of DLBCLs, and 80% of LPLs This is of great importance since HCLs and LPLs are rarely abnormal by conventional cytogenetics using the more traditional combinations of mitogens making it difficult to identify markers of prognostic significance. In conclusion, our findings demonstrate that the DSP30/IL2 cocktail induces proliferation of various B-cell mature lymphoid disorders and that its mitogenic action is not limited to CLL. We are continuing to develop our understanding of the considerable response of specific lymphoid malignancies to the DSP30/IL2 cocktail by correlating additional clinical data, and hope that the end result will open new avenues in regards to prognostic outcome and therapeutic approaches. Disclosures: No relevant conflicts of interest to declare.

Description: Higher levels of procoagulant factors and factor XII deficiency may be risk factors for first venous thromboembolism (VTE). We studied associations of coagulation factors IX through XIII with risk of future VTE in 2 general population samples. Using a nested case-control study combining the 21 860 participants of the Atherosclerosis Risk in Communities study and the Cardiovascular Health Study, we determined antigenic levels of these coagulation factors in primarily pre-event blood samples from 462 participants who subsequently developed VTE and 1047 participants who remained free of VTE. Only elevated levels of factors IX and XI were associated with increased risk of VTE after adjustment for age, sex, race, and study. For factor IX, the odds ratio (OR) was 1.4 (95% confidence interval [CI], 1.0-2.0) comparing the top to bottom quintile. The OR for factor XI was higher: 2.0 (95% CI, 1.4-2.9). With further adjustment for body mass index and diabetes, only elevated factor XI remained associated with VTE risk: OR 1.8 (95% CI, 1.3-2.7). Associations were similar by study and whether the thrombosis was idiopathic or secondary. Factor XII deficiency was not related to VTE risk. Among these procoagulant factors, only elevated factor XI was a risk factor for VTE.

Description: The concept of endothelial progenitor cells (EPCs) has attracted considerable interest in cardiovascular research, but despite a decade of research there are still no specific markers for EPCs and results from clinical trials remain controversial. Using liquid chromatography–tandem mass spectrometry, we analyzed the protein composition of microparticles (MPs) originating from the cell surface of EPC cultures. Our data revealed that the conventional methods for isolating mononuclear cells lead to a contamination with platelet proteins. Notably, platelets readily disintegrate into platelet MPs. These platelet MPs are taken up by the mononuclear cell population, which acquires “endothelial” characteristics (CD31, von Willebrand factor [VWF], lectin-binding), and angiogenic properties. In a large population-based study (n = 526), platelets emerged as a positive predictor for the number of colony-forming units and early outgrowth EPCs. Our study provides the first evidence that the cell type consistent with current definitions of an EPC phenotype may arise from an uptake of platelet MPs by mononuclear cells resulting in a gross misinterpretation of their cellular progeny. These findings demonstrate the advantage of using an unbiased proteomic approach to assess cellular phenotypes and advise caution in attributing the benefits in clinical trials using unselected bone marrow mononuclear cells (BMCs) to stem cell-mediated repair.

Description: The metalloprotease ADAMTS13 (a disintegrin and metalloprotease with thrombospondin motif) converts the hyperreactive unusually large (UL) forms of von Willebrand factor (VWF) that are newly released from endothelial cells into less active plasma forms by cleaving a peptide bond in the VWF A2 domain. Familial or acquired deficiency of this metalloprotease is associated with thrombotic thrombocytopenic purpura (TTP). ADAMTS13 belongs to the ADAMTS metalloprotease family, but, unlike other members, it also contains 2 C-terminal CUB domains (complement component Clr/Cls, Uegf, and bone morphogenic protein 1). Mutations in the CUB region have been found in congenital TTP, but deletion of the region did not impair enzyme activity in conventional in vitro assays. We investigated the functions of the CUB domain in ADAMTS13 activity under flow conditions. We found that recombinant CUB-1 and CUB-1+2 polypeptides and synthetic peptides derived from CUB-1 partially blocked the cleavage of ULVWF by ADAMTS13 on the surface of endothelial cells under flow. The polypeptide bound immobilized and soluble forms of ULVWF, and blocked the adhesion of ADAMTS13-coated beads to immobilized ULVWF under flow. These results suggest that the CUB-1 domain may serve as the docking site for ADAMTS13 to bind ULVWF under flow, a critical step to initiate ULVWF proteolysis.

Description: Severe ADAMTS13 deficiency in thrombotic thrombocytopenic purpura (TTP) is either constitutional and caused by ADAMTS13 mutations, or acquired and most often due to ADAMTS13 inhibitory autoantibodies. In strongly hemolytic serum of a pediatric patient, diagnosed with TTP postmortem, ADAMTS13 activity was less than 3%. Both parents had an ADAMTS13 activity of approximately 50%. Sequencing of the ADAMTS13 gene revealed an intronic 687-2A〉G substitution affecting exon 7, homozygous in the propositus and heterozygous in both parents, confirming constitutional ADAMTS13 deficiency. ADAMTS13 activity of normal plasma was inhibited by incubation with the propositus' serum, suggesting alloantibody formation to ADAMTS13. However, immunoglobulin purified from serum had no ADAMTS13 inhibitory effect, whereas the immunoglobulin-depleted hemolytic serum inhibited ADAMTS13 activity of normal plasma, suggesting an inhibitory effect of hemolysis products. Incubation of hemoglobin, recombinant and from lysed erythrocytes, with normal plasma revealed an ADAMTS13 inhibitory effect at hemoglobin concentrations of 2 g/L or higher.

Description: Donor cell expression of C3 enhances the alloimmune response and is associated with the fate of transplantation. To clarify the mechanism for enhancement of the immune response, we have explored the role of C3a receptor (C3aR)–ligand interaction on murine bone marrow dendritic cells (DCs). We show that DCs either lacked receptor for C3a (a C3 cleavage product) or were treated with C3aR antagonist, elicited defective T-cell priming against alloantigen expressed on the DCs. This was associated with reduced surface expression of major histocompatibility complex (MHC) and costimulatory molecules on the DCs, and with defective priming in skin allograft rejection. In addition, DCs lacking factor B were unable to generate potent T-cell responses against donor antigen, whereas lack of C4 had no detectable effect, suggesting a role for the alternative pathway contributing to allostimulation. Furthermore, therapeutic complement regulator can down-regulate DC allostimulatory function. These findings suggest that the capacity of DCs for allostimulation depends on their ability to express, activate, and detect relevant complement components leading to C3aR signaling. This mechanism, in addition to underpinning the cell-autonomous action of donor C3 on allostimulation, has implications for a wider range of immune responses in self-restricted T-cell priming.

Description: The role of the intrinsic coagulation system on the risk of myocardial infarction is unclear. In the Study of Myocardial Infarctions Leiden (SMILE) that included 560 men younger than age 70 with a first myocardial infarction and 646 control subjects, we investigated the risk of myocardial infarction for levels of factor XI (factor XIc) and factor XII (factor XIIc). Furthermore, the risks for factor VIII activity (factor VIIIc) and factor IX activity (factor IXc) were assessed. Factor XIc was 113.0% in patients compared with 109.8% in control subjects (difference, 3.2%; 95% CI, 1.1%-5.4%). The risk of myocardial infarction adjusted for age for men in the highest quintile compared with those in the lowest quintile was 1.8-fold increased (ORadj, 1.8; 95% CI, 1.2-2.7). In contrast, factor XIIc among patients with myocardial infarction was lower than in control subjects, respectively, 93.0% and 98.6% (difference, 5.6%; 95% CI, 3.3%-7.9%). The odds ratio of myocardial infarction for men in the highest quintile versus those in the lowest quintile was 0.4 (ORadj, 0.4; 95% CI, 0.2-0.5). The highest risk was found among men with both high factor XIc and low factor XIIc (analyses in tertiles: ORadj, 6.4; 95% CI, 2.2-18.0). Factor VIIIc increased the risk of myocardial infarction although not dose dependently. Factor IXc increased the risk; odds ratio of myocardial infarction for men in the highest quintile versus those in the lowest quintile was 3.2 (ORadj, 3.2; 95% CI, 2.0-5.1). Thus, factors XIc and XIIc have opposite and synergistic effects on the risk of myocardial infarction in men; factor VIIIc and factor IXc increase the risk.

Description: Hematopoietic chimerism was analyzed in serial bone marrow samples taken from 28 children following T-cell depleted unrelated donor bone marrow transplants (UD BMT) for acute lymphoblastic leukemia (ALL). Chimeric status was determined by polymerase chain reaction (PCR) of simple tandem repeat (STR) sequences (maximal sensitivity, 0.1%). At least two serial samples were examined in 23 patients. Of these, two had evidence of complete donor engraftment at all times and eight showed stable low level mixed chimerism (MC) (

Description: Background: A variety of ophthalmologic findings have been reported in patients with anemia. Aim: To determine the effect of beta-thalassemia minor on the optic nerve head topographic analysis. Methods: A total of 39 beta-thalassemia minor patients were divided into 2 groups. Patients with iron, folic acid, or vitamin B12 deficiency were ruled out. Group 1 comprised 20 patients with anemia, and group 2 comprised 19 patients without anemia. One eye of each patient was included into the study. All subjects underwent complete ocular examination. Optic nerve head topographic analysis was performed by using a confocal scanning laser ophthalmoscope type Heidelberg retina tomograph (HRT). The following stereometric parameters were evaluated: disc area, area and volume of cup, area and volume of neuroretinal rim, measure of cup shape, and mean retinal nerve fiber layer thickness. Results: The mean age of group 1 and 2 were 26.8±7.6 and 25.6±4.5 years, respectively (P=0.91). Their mean disc areas were 2.01±0.3 mm2 and 2.53±0.6 mm2, respectively (P =0.009). The differences between groups for area and volume of cup, area and volume of neuroretinal rim, cup shape measure, and mean retinal nerve fiber layer thickness were insignificant (p〉0.05). There was no significant difference between mean intraocular pressure of both groups (p=0.93). Conclusion: In beta-thalassemia minor, patients with anemia, optic disc area showed a statistically significant reduction compared to the patients without anemia. Further clinical trials on ocular blood flow and optic nerve oxygenation changes may highlight the role anemia in the optic nerve head of the beta-thalassemia minor patients.

Description: A correlation between increase in bone markers (alkaline phosphatase (ALP) and response to bortezomib in patients with multiple myeloma (MM) has been previously described. We now report results from a prospective study examining the relationship of serum PTH variation with skeletal effects and myeloma response to bortezomib treatment. Methods: Single agent bortezomib (1.3 mg/m2 patients 1–10; 1mg/m2 patient 11–20), was administered to patients with relapsed/refractory MM on days 1, 4, 8 and 11 on a 21 day interval for a total of 3 cycles; patients were not allowed to receive concurrent bisphosphonates or any other anti-myeloma drugs during the study period. Dynamic indices of bone turnover were prospectively evaluated by high-resolution microCT. Architectural parameters such as bone volume/total volume (BVTV), trabecular number (TbN), and thickness (Tb.Th) was obtained. PTH along with bone markers (osteocalcin, calcium, magnesium, phosphorus and serum creatinine) were measured on days 1, 4, 8, 11 before and after each bortezomib dose and every 4 hours thereafter, daily for the other days of the treatment cycle. Results: Seventeen patients were enrolled in the study with a median age of 63 years, 41 % were male and 3/4 of the patients previously failed high-dose chemotherapy. Histomorphometric microCT comparative analysis was completed (baseline and post-treatment) in 7 of the 17 patients enrolled in the trial. Baseline BV/TV values ranged from 13% to 90%. After 3 cycles of bortezomib treatment a statistically significant increase in BV/TV was recorded in 6 of 7 patients (P

Description: We identified a dog with large granular lymphocytic leukemia and cutaneous lymphoma that exhibited constitutive expression of interleukin-2 (IL-2) receptors by the leukemic peripheral blood lymphocytes. The leukemic cells phenotypically resembled natural killer (NK) cells, and their surface IL-2 receptors were functional, as determined by the capacity to bind human recombinant IL-2 with high- affinity resulting in the transduction of proliferation signals and in the development of lymphokine-activated killer cell activity. These cells produced IL-2 spontaneously, and they may have maintained their proliferative state through an IL-2-dependent autocrine growth pathway. Our results indicate that neoplastic lymphocytes of syndromes that involve circulating leukemic cells with dermotropism can originate from NK-like cells. Additionally, the data also suggest that proliferative conditions such as these may be the result of the aberrant production of IL-2. Further, this case illustrates the potential for the use of hematopoietic malignancies in the dog as a suitable animal model for immune targeting of IL-2 receptors as a novel treatment approach for similar malignancies of human beings.

Description: Abstract 18 Introduction: Long-term survival in pediatric relapsed AML is only 20-30%. Optimal reinduction therapy is unknown, and there is a concern about cardiotoxicity with repeated anthracycline use at relapse. Preclinical in vitro and animal studies, and limited clinical data suggest that liposomal daunorubicin (DaunoXome®, DNX) is less cardiotoxic. These considerations lead to a phase III study, in the setting of the International BFM Study Group. Materials and methods: FLAG was randomised against FLAG/DNX in the 1st reinduction course. The conventional 5-days FLAG only was recommended as the 2nd course. DNX was dosed at 60 mg/m2/day on days 1, 3 and 5. After induction, allogeneic stem cell transplantation was generally recommended, but time-to-transplant could be bridged by high- or low-intensive consolidation therapy. Primary endpoint of the study was early treatment response, based on bone marrow examination shortly before reinduction course 2, and defined as either good (≤20% leukemic blasts) or poor (〉20% leukemic blasts). This endpoint was chosen because of its prognostic value in earlier relapsed AML BFM-trials, and because compliance with an extended protocol guideline was likely to be suboptimal within the context of a highly multinational and multicenter AML Relapse protocol. However, secondary endpoints were defined, including the CR2 rate determined after 2 courses, long-term survival, and toxicity. Patients with AML M3 and those 〉18 years of age at initial diagnosis were ineligible. The study opened in most countries in 2002/2003. The study closed for accrual on April 1, 2009 when the required 360 fully eligible and evaluable patients had been randomized. Early and late relapsed AML was defined as a relapse within or after 1 year from initial diagnosis, but this only influenced treatment in that early relapsed AML patients were eligible for haploidentical SCT, while late relapsed AML patients were eligible for autologous SCT, if a matched or partly mismatched transplant was not possible. Thirteen groups from 20 countries and 〉100 centers have enrolled patients, with informed consent and after approval of the study by regulatory authorities. Data are presented according to intention-to-treat, with a median follow-up of 2.7 years for patients at risk. Results: Overall 4-year probability of survival (pOS) was 35% SE 2%, the overall CR2 rate 62%. The good early responders had a 4-year pOS of 45% SE 3% versus 10% SE 3% for poor responders (p