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: 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: 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: 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: 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: 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: 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: 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: 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: 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: GA101 is a novel monoclonal antibody of IgG1 type which binds with high affinity and selectivity to the extracellular domain of the human CD20 antigen on B cells. In contrast to rituximab which is a chimeric antibody and recognizes a type I epitope, GA101 is humanized and recognizes a type II epitope which is also localized in the extracellular loop of CD20. The recognition of the type II epitope together with a modification of the elbow hinge region results in enhanced direct non-caspase dependent cell death induction, and concomitant reduction in CDC upon binding to CD20. In addition, using GlycoMab technology, the Fc-region of GA101 was glycoengineered to contain bisected, afucosylated carbohydrates. As a result GA101 has increased affinity for the low and high affinity FcγRIIIa receptor expressed on natural killer cells, macrophages and monocytes. Consequently, GA101 mediated a 5–50 fold enhanced induction of effector cell mediated ADCC. In B-cell depletion assays with whole blood from healthy donors, an assay combining all mechanisms of action, GA101 was significantly more potent and efficacious in depleting B cells than rituximab. In preclinical NHL testing these properties translated into superior anti-tumoral efficacy of GA101 in direct comparison to rituximab against a number of aggressive NHL xenograft models. In cynomolgus monkeys the induction of B cell depletion mediated by GA101 and subsequent B cell recovery were investigated. GA101 induced complete, rapid and long-lasting B cell depletion both in peripheral blood and in lymphoid tissue e.g. spleen and lymph nodes. The efficacy of GA101 (10 and 30 mg/kg) at depleting B cells in different lymphoid tissues of cynomolgus monkeys was compared with that of rituximab (10 mg/kg) following 2 i.v. doses administered on days 0 and 7. Notably, GA101 showed statistically superior depletion of total B cells from lymph nodes compared to Rituximab from day 9 to 35 onwards with B cell numbers decreased by over 95%. These results demonstrated that GA101 was more efficacious at depleting B cells from lymph nodes and spleen of cynomolgus monkeys compared to rituximab. Compared to existing antibodies, GA101 constitutes the first type II CD20 antibody engineered for increased ADCC with significantly enhanced efficacy in a variety of preclinical models. Based on these data it is assumed that the combination of the recognition of a type II epitope together with improved ADCC potency might translate into superior efficacy in the clinical treatment of CD20 positive malignant diseases.

Description: Polycythemia vera (PV) and essential thrombocythemia (ET) are two common types of myeloproliferative disorders (MPD). The prevalence of PV and ET in the United States (US) has not been well documented. Recent breakthroughs in the molecular etiology of these disorders and the accelerated development of targeted pharmacotherapeutics to treat the MPD have significantly increased the need to accurately define the affected population. In the present study, we obtained detailed demographic and health claims data from major commercial insurance payers in Connecticut and the Center for Medicare and Medicaid Services to estimate the prevalence of PV and ET. Health claims data from one payer and the actual diagnoses made by physicians who submitted claims with MPD-related ICD-9 codes were utilized to develop claim-based statistical algorithms to predict the probability that an individual with claims suggestive of MPD truly has PV or ET. Specifically, logistic regression was used to develop the algorithms, and area under the receiver operating characteristics curve (AUC) was used as the measure of goodness-of-fit for each model. Different models were fit, and the model with the highest AUC was selected. For PV, the best-fitting model included age as a continuous variable and the frequency of two PV-related ICD-9 codes (238.4 and 289.6 combined) as a continuous variable, and the AUC was 0.95. For ET, the best-fitting model included age as a continuous variable and the frequency of code 289.9 as a categorical variable (=0, =1, or ≥2), and the AUC was 0.72. For both models, the addition of gender or the frequency of the non-specific code 238.7 did not improve AUC. Subsequently, the algorithms were applied to health claims from multiple payers to estimate the number of PV and ET patients in Connecticut. The total number of Connecticut residents included in the study was close to 2,900,000, which represented about 83.2% of the estimated population of the entire state in July 2003. As of 2003, the age-standardized prevalence of PV and ET in Connecticut was 22 per 100,000 and 24 per 100,000, respectively. Applying the age-specific prevalence of PV and ET to the entire US population resulted in an estimated total of 65,243 patients with PV and 71,078 patients with ET in the US in 2003. This study is the first to assess the prevalence of PV and ET in a very large US population. Given the large number of individuals afflicted with these diseases and the fact that demographic changes alone will further increase the burden of these diseases in the foreseeable future, it is imperative to conduct more systematic research into the etiology and treatment of PV and ET.

Description: Background: Neurologic complications of multiple myeloma (MM) are numerous, however little is known about strokes occurring in the course of MM and its treatment. Methods: 2877 MM patients were seen for an initial evaluation at our institution over a 5-year period (1998–2002). Patients were identified using discharge summaries by combining ICD-9 codes for MM and stroke. They were included if they had clinical and radiological features of acute stroke. Ischemic stroke was defined as a new acute neurologic deficit lasting 〉 24 hours, irrespective of diffusion-weighted MRI results. Diagnosis of hemorrhagic stroke was made in patients with new acute neurologic deficit plus evidence of hemorrhage on CT scan. We retrospectively reviewed medical records for demographics, type of MM and treatment, stroke type, clinical features, relevant imaging and laboratory data, and outcomes. Results: The 11 patients that we identified included eight with ischemic strokes and three with subarachnoid hemorrhages (SAH). There were no patients with intraparenchymal hemorrhage. The overall incidence of stroke in this cohort was 76/100,000 per year, whereas the incidence of ischemic stroke was 56/100,000 per year. The mean age of patients with ischemic stroke was 59 years. Seven had one or more stroke risk factors and an equal number had received thalidomide. The latter were given small doses of coumadin as part of the treatment protocol; at the time of the stroke, INR range was 0.9–1.4. Based on clinical, radiological and other laboratory features, of the eight patients with ischemic strokes, two were presumed cardio-embolic (one had infective endocarditis), five were thrombotic, and one was caused by hypoperfusion. Two patients had findings of severe stenosis/occlusion in vessels corresponding to the infracted brain territory (internal carotid and basilar arteries). Two patients had documented normal plasma viscosity and three had evidence of extra-cranial thrombosis at the time of the stroke. In the three patients with SAH, hemorrhage occurred in the setting of trauma and thrombocytopenia, although one patient had an incidental anterior communicating artery aneurysm. Seven patients were left with minor or no deficits and four died, two in each of the groups. Conclusion: Overall, strokes did not appear to be more common in MM patients than in the general population, and the pathophysiology was likewise, not different than in patients without MM, with the exception of one patient with infective endocarditis, which is probably directly related to MM or its treatment. The highly selected referral population at our myeloma institute, and the retrospective nature of the study probably underestimate the incidence of stroke. The role of thalidomide in stroke remains unknown. Thrombocytopenia and trauma are probable risk factors for subarachnoid hemorrhage in MM patients. Fatal outcomes were frequent, but when death did not occur, the deficits were minor.

Description: Recent advances in the diagnosis, molecular pathogenesis, classification and therapy have been made in the field of myelodysplastic syndrome (MDS) and juvenile myelomonocytic leukemia (JMML) in childhood. We report a retrospective analysis of children with MDS and JMML diagnosed between 2001 and 2006 in Korea. In total, 135 patients were enrolled from 19 major hospitals with pediatric hematology oncology clinics: MDS, 96 (primary MDS, 77; constitutional anomalies with MDS, 13; treatment-related MDS, 6) and JMML, 39. The incidence of MDS/JMML was around 22.5/year, which is about 6% of childhood leukemia. Various classification systems including FAB, WHO, IPSS, CCC system, and pediatric adjustment of the WHO classification were applied. The median ages at diagnosis were 68 and 10 months in MDS, and JMML, respectively. Males dominated in JMML. Cytogenetic abnormalities were observed in 43% of MDS (monosomy 7, 5; trisomy 8, 3) and in 10% of JMML. Treatment was chosen by each institute’s preference: 34 patients with MDS received AML-type intensive chemotherapy, with complete remission rate of 82.0%. The 5-year Kaplan-Meier overall survival rate was 54% each for MDS and JMML. Survival of MDS patients was influenced by the marrow blast % (P = 0.007) and disease category (P= 0.006). Stem cell transplantations (SCT) were undertaken in 56 patients (MDS, 29; JMML, 27). The sources of stem cells were: bone marrow, 36; umbilical cord, 18; peripheral blood, 2). Matched related transplants were 9 cases. Conditioning was various, but BuCy based regimen was used in 68.4%. Acute GvHD ≥ Grade II was found in 43.8% and chronic GvHD in 35.1%. The 5-year Kaplan-Meier overall survival rate was 55% for MDS, and 57% for JMML. Survival after unrelated transplant was comparable with that of matched related transplants. This analysis inspired the necessity of nation-wide prospective studies in Korea, including morphologic study by a central pathology review board, epidemiologic study, molecular pathophysiologic study, and therapeutic trials incorporating SCTs, or new drugs.

Description: The somatic mutation JAK2 V617F has been identified as a pathogenic factor in typical chronic myeloproliferative diseases (MPD) such as polycythemia vera (PV), essential thrombocythemia (ET), and myelofibrosis with myeloid metaplasia (MMF). In typical forms of myelodysplastic syndromes (MDS), JAK2 V617F mutation is rarely present (2–5%); on the contrary, it has been found with higher prevalence in patients with RARS-T (i.e. MDS/MPD-U with platelet count 〉600×109/L and ringed sideroblasts more than 15%) and in a subgroup of MDS patients with isolated 5q deletion and a proliferative bone marrow. In this study we analysed the JAK2 V617F mutational status in 53 MDS patients (26 males, 27 females; median age at the time of the study 76 years, range 45–91). Patients were classified as follows: 4 cases 5q- syndrome, 3 RCMD, 5 MDS/MPD, 1 MDS-U, 23 RA, 12 RARS, 5 RAEB. DNA was extracted from purified granulocytes; all samples were analyzed by allele-specific polymerase chain reaction (PCR), according to Baxter el al (2005). DNA samples were further subjected to direct sequencing for confirmatory testing. The JAK2 V617F mutation was present in 3 cases, with an overall frequency of 5%. With respect to MDS subtype, 1 patient had RA and 2 RARS. Among the 12 RARS patients, the two V617F postive displayed thrombocytosis (680×109/L and 649×109/L), whereas none of the 10 RARS V617F negative patients showed high platelet counts (median Plt 157×109/L, range 5–422×109/L). In one JAK2 mutant case, thrombocytosis required treatment with hydroxyurea. Moreover, the two V617F positive RARS patients displayed higher WBC count (6.2×109/L and 8.5×109/L) than the V617F negatives (median WBC 4.05×109/L); no difference was observed in Hb levels. The JAK2 positive RA patient had 10% of sideroblasts in bone marrow, normal platelet and WBC count and no proliferative characteristics; since the occurrence of the mutation may be an early event and preceed the classical manifestations of MDS/MPD, a longer follow-up is necessary to determine its possible prognostic significance. Considering the V617F negative MDS cases, only one patient, diagnosed as MDS/MPD, showed a platelet count 〉600×109/L. In conclusion, we confirmed recent reports showing that JAK2 V617F is present with low prevalence (about 5%) in MDS; in particular, the JAK2 mutation identifies a subset of MDS patients with and “overlap” syndrome, characterised by proliferative bone marrow morphology and frequent thrombocytosis and leucocytosis, who may benefit from JAK2 specifically targeted therapies.

Description: The simultaneous manifestation of different lymphomas in the same patient or in the same tissue is defined composite lymphoma. Although reports of synchronous or metachronous Hodgkin’s lymphoma (HL) and Non Hodgkin’s lymphoma (NHL) are not uncommon in the literature, the biologic relationship of the 2 malignancies is often unclear. Primary cutaneous B-cell lymphomas (pCBCLs) have been recognized as distinct clinicopathologic entities; they represent a wide spectrum of lymphoproliferative disorders separated from of B-cell NHL secondarily involving the skin and cutaneous B-cell pseudolymphomas. As regarding pCBCLs, a concomitant diagnosis of HL has been described very rarely. This is the case of a caucasian man affected by primary cutaneous follicolar B cell lymphoma (pCFCL). He presented grouped red plaques located on the nape, abdomen, shoulders, arms and even some little tumors surrounded by erythematous papules. Complete staging procedures did show no evidence of extracutaneous disease. A subcutaneous interferon therapy was started, in some months the patient reached a complete remission, and a maintainance therapy was continued for about 2 years. After 15 years, at the age of 58, the patient presented a red to violaceous infiltrated solitary plaque on the back, appeared about 2 months before. The lesion was completely excised and the biopsy showed a diffuse dermal infiltrate, not involving the epidermis, structured in follicle with reactive germinal centers; they were surrounded by small sized monomorphic lymphocytes with irregular nuclei and pale cytoplasm, showing the following immunophenotype pattern: CD20+, CD3−, IRTA+, CD10−, BCL6−, BCL2+, low proliferative index. A plasmacellullar CD138+ and CD79a+ population was at the periphery of the infiltrates, with monotypic expression of cytoplasmic k chain. The whole picture was interpreted as primary cutaneous marginal zone B-cell lymphoma (pCMZL). The association with Borrelia burgdorferi infection, sometimes described in pCMZL, wasn’t demonstrated. The patient presented a large right axillary lymph node that was excised and found to be unexpectly infiltrated by HL, mixed cellularity subtype. The patient underwent a standard baseline staging procedure with total body CT scan and bone marrow trephine biopsy; the latter resulted negative; the t(11;14) and t(14;18) rearrangements weren’t demonstrated in bone marrow; the CT did show no other suspected masses nor lymphoadenopathy, besides the clinically evident right axillary lymph node. A 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) revealed sites of hyperactivity in the same right axillary region, but extending to the subclavian region and the thoracic region. The patient started chemoterapy (ABVD regimen), and at the end of 4 courses a whole body CT scan and FDG-PET resulted both negative. Now he is being treated with radiotherapy. If the nodal malignancy would have been diagnosed first, the skin lesion probably could have been misinterpreted as a secondary localization of HL; if the node biopsy wouldn’t have been performed we could diagnose B-cell NHL secondarily involving the skin (stage IV). Nevertheless the cutaneous and nodal infiltrates had a completely different general picture and phenotype. This case probably reflects a HL after 15-year remission of a pCBCL of low grade and the relationship between HL and the preceding pCFCL is not clear: casual or related to genetic predisposition for oncogenic events or favoured by an immunodeficiency state related to the first disease and the previous immunomodulatory therapy.

Description: Tyrosine kinase inhibitors (TKI) selective for Bcr-Abl, such as dasatinib, imatinib, and nilotinib have had remarkable success in the clinic, potentially shifting the prognosis of chronic myelogenous leukemia (CML) to a manageable chronic disease. With the increase in longevity of CML patients, there is rising concern of co-morbidities that may be influenced by chemotherapy (Force et al., Nature Rev.2007;7:332–340). Recently, congestive heart failure (CHF) and direct cellular cardiotoxicity have been reported in CML patients on imatinib therapy (Kerkela et al., Nature Medicine2006;12:908–916). Ultrastructural mitochondrial abnormalities in cardiomyocytes were observed in CML patients with severe CHF and, interestingly, similar abnormalities were observed in cardiomyocytes of imatinib-treated mice, thus providing a prospective in vivo animal model for imatinib-induced cardiotoxicity. Furthermore, correlative findings of mitochondrial membrane potential loss, decreased cell viability, and increased apoptosis resulted from an array of cell-based assays in imatinib-treated primary rat cardiomyocytes, consequentially affording a supportive, if not predictive, in vitro cardiomyocyte toxicity model. Since imatinib-induced inhibition of the native form of c-Abl kinase was speculated to cause the observed cardiotoxicity and c-Abl is a shared target of dasatinib, imatinib, and nilotinib, the in vitro cardiotoxicity potential of dasatinib and nilotinib at pharmacologically relevant concentrations (0.09 μM and 5 μM, respectively) and up to 10-fold higher concentrations were compared side-by-side with imatinib in primary rat cardiomyocytes. Dasatinib did not significantly affect mitochondrial membrane potential, cell viability, apoptosis, or cellular ultrastructure in vitro, whereas imatinib significantly affected these parameters. Nilotinib at pharmacologically relevant concentration demonstrated decreased cell viability, but differed from imatinib in that mitochondrial membrane potential integrity was not affected under identical experimental conditions. Results suggest that at pharmacologically relevant concentrations, dasatinib does not induce cardiotoxicity, as does imatinib and nilotinib, and the molecular mechanisms of the observed cardiotoxicities may differ between imatinib and nilotinib. Of indirect relation, results from assessing another cardiovascular liability, namely hERG K+ channel blockade, demonstrated that dasatinib, imatinib and nilotinib differentially inhibited the hERG currents in vitro with IC50 of 14.3, 15.6 and 0.66 μM, respectively. These in vitro findings occurred at concentration levels approximately 150, 3 and 0.1-fold the expected human Cmax for the three TKIs, respectively. Thus, although TKI therapies may share similar targeting and clinical indications, differentiating specific toxicity profiles may be predictive of differences in potential clinical adversities.

Description: Factors regulating which patients become alloimmunized to red blood cell (RBC) antigens are poorly understood. Using a murine model of transfusion, we recently reported that viral-like inflammation with polyinosinic polycytidylic acid [poly (I:C)] significantly enhances RBC alloimmunization. Herein, we tested the hypothesis that poly (I:C) exerts this effect, at least in part, at the level of antigen-presenting cells (APCs). Using a novel in vivo method, we report that in the noninflamed state, most transfused RBCs were consumed by splenic macrophages, with only trace consumption by splenic dendritic cells (DCs). To a lesser extent, RBCs were also consumed by APCs in the liver. However, unlike soluble antigens, no RBCs were consumed by APCs in the lymph nodes. Inflammation with poly (I:C) induced significant consumption of transfused RBCs by splenic DCs, with a concomitant increase in costimulatory molecule expression. Moreover, this resulted in increased proliferation of CD4+ T cells specific for the mHEL RBC alloantigen. Finally, splenectomy abrogated the enhancing effects of poly (I:C) on RBC alloimmunization. Together, these data provide additional insight into the nature of transfused RBCs as an immunogen and provide a mechanism by which viral-like inflammation enhances alloimmunization to transfused RBCs.

Description: Dietary plant polyphenols are known to have antitumor, antiinflammatory, and antioxidant activity and as such may sensitize tumor cells to chemotherapeutic agents. We have evaluated the effects of the green tea polyphenol epigallocatechin gallate (EGCG) alone and in combination with other drugs on human myeloma cells. The compound is currently under investigation in several phase I/II clinical trials including for treatment of patients with early stage chronic lymphocytic leukemia. EGCG inhibited the in vitro growth of human myeloma cell lines in a time and dose-dependent manner. IC50 concentrations were between 12.5 μM and 50 μM as measured in a colorimetric tetrazolium (MTS) based assay and by trypan blue exclusion. Excess amounts of IL-6, bone marrow stromal cells, or overexpression of Mcl-1 and Bcl-xL could not protect from EGCG induced cytotoxicity. Pretreatment of INA-6 cells with EGCG resulted in a dose-dependent inhibition of IL-6 induced STAT3 tyrosine phosphorylation. In accordance with the essential role of STAT3 for INA-6 cell survival, EGCG induced apoptosis as determined by flow cytometry upon 7-amino-actinomycin D/annexin-V staining. In cell lines not dependent on exogenous IL-6, EGCG induced growth inhibition was abolished by pretreating the cells with 200 U/ml catalase, an enzyme which reduces reactive oxygen species (ROS). The combination of EGCG with doxorubicin, dexamethason, or rapamycin did not result in increased growth inhibition. In contrast, growth inhibition by bortezomib was antagonized with EGCG at concentrations that were not inhibitory when used alone (1–10 μM). In conclusion, EGCG exerts its effects on myeloma cells through several mechanisms including inhibition of IL-6 mediated signalling pathways via STAT3 and induction of oxidative stress. Notably, at pharmacologically achievable concentrations, EGCG antagonized bortezomib activity. Thus, the intake of natural polyphenols (high consumption of green tea or taking green tea extracts) may be critical during therapy with bortezomib.

Description: PU.1 (Sfpi1) is an ets family transcription factor required for the proper generation of both myeloid (macrophages and neutrophils) and lymphoid lineages (B and T lymphocytes)(Scott 1994, McKercher 1996). Graded expression of exogenous PU.1 in murine PU.1-deficient fetal liver hematopoietic progenitors demonstrated that increased levels of PU.1 are required to initiate development of macrophages (DeKoter, 2000). We have studied the effects of graded expression of PU.1 on its occupancy in chromatin and on the development of myeloid cells in vitro. We measured changes in gene expression, PU.1 occupancy and histone modifications in PU.1-null hematopoietic progenitor cells stably expressing PU.1 fused to the ligand-binding domain of the estrogen receptor (PU.1-ER) (Walsh 2002). The level of active PU.1-ER was regulated with graded levels of the ER inducer tamoxifen. In vitro, intermediate levels of tamoxifen produced cells with granulocyte characteristics in the suspension cell fraction and macrophage-like characteristics in the attached fraction, whereas high levels of PU.1 produced mostly attached macrophage-like cells. Expression of granulocyte-specific PU.1 target mRNAs including gelatinase B (Mmp9) and myeloperoxidase (Mpo) were observed to be expressed only with intermediate levels of tamoxifen. In contrast, expression of macrophage PU.1 target mRNAs including Cd14, F4/80 and Cd68 mRNAs were observed to be gradually upregulated upon PU.1-ER activation, with the maximum expression at the highest levels of tamoxifen. Thus, the expression levels of PU.1 target genes and phenotypic characteristics of the cells are dependent on PU.1 levels. Interestingly, macrophage-like cells can be produced from granulocytic-like cells by changing tamoxifen levels and vice versa. Chromatin immunoprecipitation analysis revealed specific PU.1 occupancy within regulatory regions of the genes predominantly expressed in macrophages including Cd14 and Cd11b after treatment with high levels of tamoxifen. Specific PU.1 occupancy within regulatory regions of the granulocyte specific genes including MMP9 was observed at intermediate levels of tamoxifen. Suprisingly, chromatin immunoprecipitation analysis revealed specific PU.1 occupancy within regulatory regions of the lymphocytic PU.1 target genes including Interleukin-7 receptor (Il-7r) and RAG1 at intermediate levels of tamoxifen even though expression of these genes was not detected. Accumulation of acetylated K9 and methylated K4 of histone H3 in gene loci of macrophage and granulocytic markers such as Cd14, Cd11b, and Mmp9 correlated with their mRNA expression. However, lymphocyte-specific regulatory regions including that of Il-7r gene were hypoacetylated in H3K9 despite a marked PU.1 recruitment suggesting additional factors may be required for PU.1 mediated transactivation. To identify these molecules we have tested PU.1-dependent transcription factors: Egr2, Nab2, Cebpa and Gfi-1 and found that upon increasing PU.1 levels, expression of Egr2/Nab2 and Gfi-1/Cebpa changed in a reciprocal manner and these changes preceded expression of the lineage specific markers. We are currently testing if PU.1 directly regulates expression of Egr2, Nab2, Cebpa and Gfi-1 during granulocytic/macrophage differentiation.

Description: CD26 (dipeptidyl peptidase IV, DPP IV) is a multifunctional type II cell surface glycoprotein that is widely expressed on T and natural killer cells, as well as on epithelial, endothelial and acinar cells of different tissues; its expression on B cells is very low but it is greatly upregulated following activation. We evaluated, by means of flow cytometry, the expression of CD26 in various types of B-cell lymphoid tumors: Follicular Lymphoma (Fo-Ly, 12 cases), Mantle cell Lymphoma (MCL, 12 cases) Multiple Myeloma (MM, 20 cases), Hairy cell Leukemia (HCL, 12 cases), B-cell Chronic Lymphocytic Leukemia (B-CLL, 112 cases), CD5 negative B-cell Chronic Lymphoproliferative Diseases (CD5neg-B-CLPD, 20 cases) and Diffuse Large cell Lymphoma (DLCL, 12 cases). CD26 expression was absent or low of Fo-Ly and MCL, high on MM and HCL, variable on B-CLL, CD5neg-B-CLPD and DLCL. Fluorescence intensity of positive cells was dim in B-CLL and CD5neg-B-CLPD, heterogeneous in DLCL, and bright in HCL and MM. Interestingly, in CLL patients, CD26 expression was significantly correlated with CD49d and CD38 expressions (p

Description: Due to the lack of large population-based studies, the incidence of polycythemia vera (PV) has not been well documented in the United States. In 2001, PV became reportable to the Surveillance, Epidemiology, and End Results (SEER) program, which consists of high-quality population-based cancer registries supported and sponsored by the National Cancer Institute. In the present analysis, we accessed the SEER data to estimate the incidence of PV in the United States during 2001 – 2003. Since the addition of PV to SEER reporting is fairly recent, and the diagnosis of PV is different from that of many other types of cancer, especially solid tumors, under-reporting is possible. Therefore, we also used health claims data from Medicare services to estimate the incidence of PV among individuals 65 years and older. SEER data suggest that the age-adjusted incidence rate of PV was 0.87 per 100,000 per year (95% confidence interval 0.83 – 0.91) during 2001 – 2003, which is lower than the estimates from other studies (Ania BJ et al. Trends in the incidence of polycythemia vera among Olmsted County, Minnesota residents, 1935–1989. Am J Hematol1994;47(2):89–93. Johansson P. Epidemiology of the myeloproliferative disorders polycythemia vera and essential thrombocythemia. Semin Thromb Hemost2006;32(3):171–3). The incidence of PV increased with age and was higher in men and white individuals than in women and other racial groups. The SEER-derived age-adjusted incidence rate of PV was 4.1 per 100,000 per year among individuals 65 years and older during 2003, which is considerably lower than the estimates made from nationwide Medicare claims data in the same age group during the same year, suggesting possible under-reporting to SEER. The addition of PV to SEER reporting represents a major step in generating population-based epidemiologic data for this understudied malignancy. However, it is critical to ensure the completeness in the ascertainment and reporting of PV so high-quality SEER data can be utilized to facilitate much needed research on the etiology and outcomes of PV, as well as other myeloproliferative disorders.

Description: Introduction: Filgrastim is widely used for mobilizing CD34+ cells into the peripheral blood that are easily collected by apheresis for allogeneic transplantation. With case reports documenting splenomegaly with life-threatening complications in normal donors, we prospectively evaluated spleen size using ultrasonography and clinical examination during PBPC mobilization and collection in a single-arm trial. Methods: Subjects ≥18 yrs eligible to be PBPC donors per institutional guidelines enrolled. Splenic assessments were done before, during, and after PBPC mobilization. Filgrastim dose and schedule and leukapheresis (LK) procedures were per institutional practice. The primary endpoint was fold change from baseline in splenic volume in post-baseline measurements during mobilization (measured by ultrasound [US]). Spleen size by US was measured in 3 dimensions similarly by all centers: longitudinal (craniocaudal), transverse, and diagonal (perpendicular to transverse in transverse image) diameters. Splenic volume was estimated by taking the cross-product of 3 dimensions and multiplying by 0.52, approximating the volume of an ellipse. Physical examination was performed on US days, assessing spleen palpability. US and palpation results were blinded from each other at assessment times. Timepoints included baseline (before first filgrastim dose), first LK (done before LK, typically day 4 or 5 of filgrastim), 2 and 4 days after first LK, and 7 days after last LK. Timepoints in the post-amendment cohort (n=219) were reduced to facilitate enrollment and were baseline and day of first LK (before LK). Results: 309 donors enrolled, median age 44yrs (range 18 to 74), 56% male. Mean daily filgrastim dose was 11.4mcg/kg (SD=3.0). Median number of LK was 1.5 (range 1 to 4). In all donors, the median increase in each measured dimension on first LK day was 1.4cm, 1.4cm, and 0.6cm (12.8%, 12.6%, and 15.0%), and the median fold volume increase from baseline to first LK was 1.47, resolving to near baseline 1 week after last LK. There was no apparent relationship between volume fold change and filgrastim dose, ANC, or CD34+ yield. Of 861 splenic palpation assessments reported in all donors, 98% were reported as nonpalpable (842 assessments), and 2% were palpable (19 assessments, 2 at baseline). Reporting of palpable spleens did not correlate with increased spleen size. Tenderness or guarding upon splenic palpation was reported in 2 donors with a spleen considered palpable and in 6 donors with nonpalpable spleens. No donor experienced a splenic rupture. Adverse events related to filgrastim were generally mild to moderate. Conclusion: During PBPC mobilization with filgrastim in normal donors, the spleen increased a median of approximately 50% from baseline to day of first LK and returned to near baseline 1 week after last LK. Size change was not associated with significant clinical sequelae. Timepoint Median fold change from baseline in splenic volume (Q1, Q3) *statistically significant (p

Description: Introduction: Fludarabine treatment has been shown to be beneficial for patients with Chronic Lymphocytic Leukemia (CLL), and fludarabine-based combinations may even further improve outcomes in patients with CLL. However, most CLL patients eventually become fludarabine refractory, a state which is associated with a relatively short survival. Treatment of fludarabine-refractory patients is challenging, with a median survival of about 10 months. Recently, 2 phase II clinical trials (Chanan-Khan et al. JCO 2006 and Ferrajoli et al. ASH 2006) demonstrated the clinical efficacy of lenalidomide, an immunomodulatory agent, in relapsed/refractory CLL patients. We conducted a subset analysis to examine the efficacy of lenalidomide in patients who are fludarabine refractory. Methods: All patients enrolled on the 2 phase II single agent lenalidomide clinical trials were evaluated and patients with fludarabine-refractory disease (progressed while on or within 6 months of fludarabine-based therapy) were assessed for clinical efficacy of lenalidomide. Lenalidomide was given orally either at 10 mg daily for 28 days followed by 5 mg increments every 28 days to a maximum dose of 25 mg or given at 25 mg on days 1–21 of each 28-day cycle. Response was assessed using the NCI-WG 1996 criteria. Results: A total of 80 patients were collectively enrolled in these clinical studies. Among these, 29 were identified to have fludarabine-refractory disease. Important clinical characteristics of these patients are reported in Table 1. The overall response rate in fludarabine-refractory patients was 34.5% (10/29). Complete remission was observed in 2 (6.8%) patients. Conclusion: Lenalidomide is a novel agent with immunomodulating properties demonstrating clinical efficacy in relapsed or refractory CLL patients. Interestingly, clinical responses to single agent lenalidomide were noted despite refractoriness to fludarabine (a subset of CLL patients with poor survival and limited therapeutic options). This observation of the clinical benefit of lenalidomide independent of responsiveness to prior fludarabine is encouraging and warrants further evaluation. Table 1 Ferrajoli et al. Chanan-Khan et al. Fludarabine-refractory (N=12) Fludarabine-refractory (N=17) ORR, overall response rate; PFS, progression-free survival; OS, overall survival. Median age, years (range) 62 (51–82) 68 (53–75) Sex, female/male 4/8 4/13 Median no. prior therapies (range) 4 (3–15) 4 (1–10) Median beta microglobulin (range) 5 (3–10) 5 (2–10) Advance Rai Stage (III/IV), n (%) 7 (58.3) 13 (76.4) ORR, n (%) 3 (25.0) 7 (41.2) Median PFS, months 12 14.9 Median OS, months All alive (range, 7–19) 23

Description: Dyskeratosis congenita (DC) is a congenital disorder characterized by very short telomeres. Clinical presentation includes a diagnostic triad (lacey reticular pigmentation, nail dystrophy, and leukoplakia), aplastic anemia (the main cause of premature death), myelodysplastic syndrome/leukemia and solid tumors. Allogeneic HCT is the only curative option for the hematologic complications of DC but has been associated with a high risk of peri-transplant morbidity and early death. Only about fifty HCT for DC have been performed to-date, and the five year survival after related donor HCT has been about 75%, but only approximately 35% when an unrelated donor was used. To improve survival in DC patients by decreasing transplant mortality, we introduced a reduced intensity regimen including cyclophosphamide (50 mg/kg), fludarabine (200 mg/kg), low dose total body irradiation (200 cGy), and (in patients 3 and 4) campath 1H (1 mg/kg). To decrease the risk of graft rejection, grafts were not T-cell depleted. We report outcomes in four consecutive patients, two adults and two children, all of whom engrafted with donor hematopoiesis: Age (years) Sex Graft and HLA match NC dose (×108/kg) CD34 dose (×108/kg) Follow-up (months) Donor chimerism 24 M URD dUCB 4/6, 4/6 0.55, 0.39 0.5, 0.43 1 (dead*) 83% 29 F REL PBSC 6/6 13.93 4.43 20 (alive) 100% 5 F URD BM 7/8 1.38 1.55 16 (alive) 100% 2 M URD BM 8/8 5.92 2.31 3 (alive) 100% Legend: M, male; F, female; NC, nucleated cell; URD, unrelated donor; REL, related donor; BM, bone marrow; dUCB, double umbilical cord blood; PBSC, peripheral blood stem cells; *Patient had autologous recovery after the first dUCB and died of sepsis 1 month after the second dUCB; HLA matching is reported for antigen level HLA-A, B and allele level DRB1 for cord blood, and allele level typing for HLA-A, B, C, DRB1 for PBSC or BM. The most recent donor chimerism is reported. To decrease the risk of graft rejection and prevent graft versus host disease (GvHD) patients received cyclosporine and mycophenolate mofetil. Patient 2 developed limited chronic GvHD and patient 4 developed grade III skin acute GvHD. Both were treated successfully with systemic and topical steroids. Our data suggest that this conditioning regimen results in a low rate of transplant related complications without compromising engraftment. Critically, early fatal pulmonary and vascular complications, common in post-transplant courses in DC patients, were not observed. This highlights the need to avoid drugs that are associated with pulmonary toxicity such as busulfan, and to limit radiation to the lung in patients with DC. This new less intensive conditioning regimen appears to result in a low rate of transplant related complications, and yet has adequate immunosuppressive activity to permit engraftment from alternative donors in DC patients.

Description: Extensive chronic graft-versus-host disease (ecGVHD) is characterized by fibrosis similar to that of patients with systemic sclerosis (scleroderma). Since stimulatory autoantibodies against the platelet-derived growth factor (PDGF) receptor (PDGFR) have been found in patients with scleroderma and are responsible for the activation of skin fibroblasts, we tested the hypothesis that these autoantibodies are also present in patients affected by ecGVHD. Serum from 39 patients subjected to allogeneic stem cell transplantation for hematologic malignancies (22 with ecGVHD and 17 without cGVHD) and 20 healthy controls was assayed for the presence of stimulatory autoantibodies to the PDGFR by incubating purified IgG with mouse-embryo fibroblasts lacking PDGFR α or β chains or with the same cells expressing PDGFR α. Stimulatory antibodies to the PDGFR were found selectively in all patients with ecGVHD but in none of the patients without cGVHD. Higher levels were detected in patients with generalized skin involvement and/or lung fibrosis. Antibodies recognized native PDGFR, induced tyrosine phosphorylation, accumulation of reactive oxygen species (ROS), and stimulated type 1 collagen gene expression through the Ha-Ras-ERK1/2-ROS signaling pathway. The biologic activity of these autoantibodies suggests a role in the development of fibrosis and argues for a common pathogenetic trait in ecGVDH and scleroderma phenotypes.

Description: Background: The initial genetic event in ∼85% of follicular lymphomas (FL), the most common B-cell lymphoma in North America, is the t(14;18)(q32;q21) resulting in over-expression of the anti-apoptotic protein Bcl-2. The secondary events associated with disease progression are not well understood. Alterations affecting the p arm of chromosome 1 are evident by standard karyotype analysis in ∼20% of FL. We have further examined the relationship between 1p deletion and FL using high resolution genomic analyses. Methods: The prevalence of 1p alterations was investigated in 139 cases of indolent and transformed FL using whole genome tiling path BAC array Comparative Genomic Hybridization (array CGH). Array-based single nucleotide polymorphism analysis was performed on a subset of cases using Affymetrix 500K SNP arrays. Results: Array CGH identified a minimum region of deletion spanning ∼0.5MB within 1p36.32 in 51 cases (37%). In 38 cases (27%) this loss was exhibited in the transformed sample but not the pre-transformation sample. The majority of cases displayed heterozygous deletion, while two cases showed homozygous deletion. The mechanisms of loss included simple deletions, unbalanced translocations with various partner chromosomes and eleven cases with an unbalanced t(1;1)(p36;q12). The Affymetrix 500 SNP array analyses showed copy neutral loss of heterozygosity or acquired uniparental disomy (aUPD) in three of ten cases that were negative for loss by aCGH. Contained within the 1p36.32 minimally deleted region are only a few candidate genes including tumor necrosis factor receptor superfamily 14 (TNFRS14), which has been implicated in growth inhibition of HT-29 human colon adenocarcinoma cells and induction of Fas-mediated apoptosis in non-Hodgkin’s lymphoma. Conclusions: Our data indicate that loss of heterozygosity at 1p36.32 through deletion or aUPD constitutes the most common secondary cytogenetic event in FL. LOH at 1p36 may represent an important step in the progression of indolent to transformed FL. Further studies have been initiated to investigate other possible gene inactivation events such as methylation and mutation.

Description: CD4+CD25+Foxp3+ regulatory T cells (CD25+ Treg cells) direct the maintenance of immunological self-tolerance by active suppression of autoaggressive T-cell populations. However, the molecules mediating the anergic state and regulatory function of CD25+ Treg cells are still elusive. Using differential proteomics, we identified galectin-10, a member of the lectin family, as constitutively expressed in human CD25+ Treg cells, while they are nearly absent in resting and activated CD4+ T cells. These data were confirmed on the mRNA and protein levels. Single-cell staining and flow cytometry showed a strictly intracellular expression of galectin-10 in CD25+ Treg cells. Specific inhibition of galectin-10 restored the proliferative capacity of CD25+ Treg cells and abrogated their suppressive function. Notably, first identified here as expressed in human T lymphocytes, galectin-10 is essential for the functional properties of CD25+ Treg cells.

Description: Background: Romidepsin is a bicyclic peptide that inhibits Class I and II HDACs. Piekarz et al noted responses to romidepsin in CTCL (ASCO, 2004). This pivotal phase II study sought to confirm the activity. Methods: This single arm, open label study enrolled CTCL (Stages 1B–1VA), including MF and Sézary syndrome (SS) patients (pts) from ∼40 sites in Europe, Russia, Ukraine, Georgia and the US. Pts with biopsy-proven CTCL (centrally reviewed) who failed ≥1 prior systemic therapy received romidepsin at 14 mg/m2 as a 4-hour IV infusion on Days 1, 8, and 15 q 28 days for up to 6 cycles but could continue if deriving benefit. Eligibility criteria included adequate organ function and ECOG PS ≤ 1. Exclusions included significant CV abnormality or treatment with QTc-prolonging or CYP3A4-inhibiting drugs. The primary endpoint is response rate measured by a combination of imaging, circulating malignant T-cell counts and a weighted scoring instrument to determine skin involvement (confirmed by photography). Target accrual of 64 evaluable pts (i.e. received 2 courses) has been reached and the study will close. Results: 92 pts were eligible with 68 evaluable for efficacy per protocol. Responses in evaluable pts are 1 CR, 3 CCRs, 20 PRs, 40 SD and 4 PD for an ORR of 35% (duration 2–21 months). Of pts who received ≥1 dose, the ORR is 26% (24/92) but includes 5 too early to be assessed. Median time to response is 8 weeks (range 4 – 20). Responses by stage at entry in evaluable pts, as available: Stage IB-IIA 7/23 (30%); Stage IIB-IVA 15/37 (41%). In pts with pruritus at baseline i.e. score of ≥ 30 mm on a 100 mm visual analogue scale (VAS), relief of ≥ 30 mm from baseline or a VAS score of 0 (no itching) for at least 2 cycles, was seen in 18/38 pts (47%). In those pts with severe pruritus (VAS score ≥70 mm), 14/24 (58%) experienced relief of itching. Adverse event (AE) data are available for 75 pts. AEs were reported in 54/75 (72%) of dosed pts but Grade (G) 3/4 events in only 12/75 (16%). Most frequent AEs are nausea/vomiting (G2) fatigue (G2/3), myelosuppression (G2/3), and asymptomatic ECG changes (transient mild QTc prolongation and nonspecific ST-T wave abnormalities). Thirteen pts withdrew due to AEs but there were no treatment-related deaths although 4 pts died of PD and 1 from right ventricular failure. Serious AEs considered possibly, probably or likely related to treatment were reported in 12 pts. Of these, 8 had ≥G3 events: tumor lysis, cardiac tamponade, sepsis, constipation, oral candidiasis, dermatitis, hyperglycemia/vomiting/nausea and bradyarrhythmia/atrial fibrillation. Conclusions: This study confirms the efficacy of romidepsin in treatment-refractory CTCL including relief of pruritus and an encouraging response rate with 4 CCR (1 pathology-confirmed). The low rate of discontinuation due to AEs and prolonged treatment duration of some patients illustrate that toxicity has been manageable.

Description: Waldenstrom’s Macroglobulinemia (WM) is an incurable B-cell malignancy characterized by bone marrow (BM) infiltration with a spectrum of clonally related cells, including small lymphocytes and lymphoplasmacytic cells (CD19+) as well as mature plasma cells (CD138+). The molecular pathogenesis of the disease remains to be defined. We therefore analyzed the gene expression profiles of CD19+ and CD138+ BM mononuclear cells from 30 untreated patients with WM and compared their gene expression profile to their normal counterparts from 10 healthy donors using Affymetrix microarrays (U133 plus 2.0). Since the microenvironment plays an important role in the pathogenesis of WM, we also profiled and compared gene expression profiling for CD19 and CD138 depleted BM mononuclear cells from the same patients and healthy donors. Gene expression analysis was performed using dChip software. Unsupervised hierarchical cluster analysis demonstrated distinct gene expression patterns between WM cells versus their normal counterparts. In supervised hierarchical cluster analysis selecting for genes with 〉 2 fold change in expression and a False discovery Rate (FDR) 〈 2%, a set of 1171, 582 and 360 genes were found to be differentially expressed between WM patient and healthy donor CD19+, CD138+, as well as CD19/CD138 depleted (microenvironmental) cells, respectively. Among the most significantly over-expressed genes in the CD19+ compartment in WM patients were: BCL2, TNFRSF13B, TNFRSF17, IGLL1, CCR2, CLLU1, whilst the AP1 family genes JUND and FOSB were among the most significantly down-regulated genes in both CD19+ and CD138+ cells in WM patients. Other interesting transcripts which were over-expressed in CD138+ cells from WM patients included those from genes involved in transcription regulation (ZKSCAN1, ZMYM1, ZNF189, ZNF19, and ZNF559) and interferon response (IFI16 and IFIH1). Of considerable interest was our observation that microenvironmental cells in WM patients demonstrated an overactive transcriptional profile composed of genes which are associated with immune and inflammatory responses including the Toll like receptors (TLR 1,5,7,8), interferon and cytokines (IFI16, IFNAR1, IL-10R, IL-8R), genes encoding extracellular matrix components (Fibronectin and Hepatocyte Growth Factor) as well as genes involved in apoptotic signaling (TNFSF10, TRAF4). These studies provide the first comprehensive molecular characterization of WM, dissecting the molecular features of the two immunophenotypically distinct populations of malignant cells, and providing for the first time evidence for a distinct molecular profile in BM microenviromental cells.

Description: Introduction: Event-free survival (EFS) for children with ALL is approximately 80%. Despite substantial success in achieving second and subsequent remissions, survival of patients with relapsed ALL (rALL) remains dismal. We propose that progress depends on identification of novel drug combinations with more activity in rALL than those commonly employed. The literature suggests a 40% CR rate for second and subsequent relapse (Br. J Haematol2005;131:579). However, CR rates depend on the number of prior therapeutic attempts and duration of any prior response, complicating identification of promising regimens. Limited patient numbers and a large number of potential candidate regimens discourage randomized trials. We surveyed local experience with rALL to establish a robust benchmark for evaluation of novel drug combinations. Methods: The TACL Consortium (www.tacl.us) was formed to develop novel drug combinations for patients with rALL. We initiated a review of all patients with rALL treated between 1995 and 2004 at eight TACL institutions. Detailed data on therapy, response, and duration of response were collected on all patients. Results: Of 313 rALL patients, 62% were males, 27% were older than 10 years at diagnosis, 26% had initial white blood counts (WBC) at diagnosis 〉=50,000/uL, and 46% were high-risk by NCI risk criteria. Re-induction attempts ranged between 1 and 9 and most commonly employed combinations of traditional ALL agents. Limiting analyses to patients with marrow involvement, we obtained 86% CR’s for 1st relapse (95% confidence interval 80%–90%), 44% for second relapse (35%–53%), and 30% for third relapse (19%–43%). CR rates declined with the number of prior treatment attempts (see Table, p

Description: Introduction: DIC is a complication that occurs during serious infection with Gram-negative bacteria. Endotoxin is a component of the bacterial cell wall that elicits a cytokine-mediated cascade of tissue factor-dependent hypercoagulable reactions. The resulting hypercoagulable state may be inhibited by potent anticoagulation. Rivaroxaban is an oral, direct Factor Xa (FXa) inhibitor in advanced clinical development for the prevention and treatment of thromboembolic disorders. The aim of this study was to examine the effects of rivaroxaban in a rat model of endotoxin-induced DIC. Methods: Rivaroxaban (0.1–10 mg/kg p.o.) or vehicle control (PEG400/H2O 60/40%, 5 mL/kg p.o.) were administered 30 minutes before endotoxin injection (lipopolysaccharide O55 B5 [LPS], 250 μg/kg i.v.) to conscious rats. Blood samples were withdrawn from anesthetized animals by heart puncture 4 hours after LPS injection and fibrinogen, platelet count, thrombin-antithrombin (TAT) complex levels and IL-6 were measured. Results: The induction of DIC was indicated in the placebo + LPS group vs vehicle control by decreased fibrinogen (2.12±0.08 vs 2.69±0.10 g/L) and platelet count (571±28 vs 904±30×109/L) an increase in TAT (75±9 vs 2±1 μg/L) and IL-6 (8.6±1.23 μg/L vs 0.028±0.020 μg/L). Pretreatment with rivaroxaban (0.1–10 mg/kg p.o.) dose-dependently ameliorated the effects of LPS on fibrinogen, platelets and TAT. Rivaroxaban 10 mg/kg p.o. normalized fibrinogen (2.58±0.07 g/L) and TAT (5.6±1.2 μg/L) and increased platelet count (703±19×109/L) (Table). Rivaroxaban also slightly reduced the plasma levels of the pro-inflammatory cytokine interleukin-6 (IL-6) (4.24±0.67 μg/L). Conclusions: These results show that direct, selective inhibition of FXa by rivaroxaban effectively normalized the hypercoagulable reactions of endotoxemia with a slight modulating effect on the generation of pro-inflammatory active cytokines, such as IL-6, in the rat DIC model. Further research into the use of rivaroxaban in the management of DIC is therefore warranted. Effects of rivaroxaban in an endotoxin-induced DIC model in rats 4 hours after LPS injection. Results show mean ± SEM. Vehicle control Placebo + LPS Rivaroxaban 1 mg/kg + LPS Rivaroxaban 10 mg/kg + LPS DIC, disseminated intravascular coagulation; IL-6, interleukin-6; LPS, lipopolysaccharide O55 B5; SEM, standard error of the mean; TAT, thrombin-antithrombin ## p