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The fine structure emission of thermospheric atomic oxygen at 63 μm wavelength was measured for the first time globally by the CRISTA (Cryogenic Infrared Spectrometers and Telescopes for the Atmosphere) experiment in November 1994. The data indicate low atomic oxygen densities throughtout the lower thermosphere during the time period of the experiment. A first analysis of the latitudinal distribution of oxygen atoms at 150 km altitude shows deviations from the respective CIRA-86 model predictions at mid to high northern latitudes. These deviations vary from day to day, and are possibly the result of changes in the geomagnetic activity.

Bioactivation of vitamin D consists of two sequential hydroxylation steps to produce 1α,25-dihydroxyvitamin D3. It is clear that the second or 1α-hydroxylation step is carried out by a single enzyme, 25-hydroxyvitamin D 1α-hydroxylase CYP27B1. However, it is not certain what enzyme or enzymes are responsible for the initial 25-hydroxylation. An excellent case has been made for vitamin D 25-hydroxylase CYP2R1, but this hypothesis has not yet been tested. We have now produced Cyp2r1 (-/-) mice. These mice had greater than 50% reduction in serum 25-hydroxyvitamin D3. Curiously, the 1α,25-dihydroxyvitamin D3 level in the serum remained unchanged. These mice presented no health issues. A double knockout of Cyp2r1 and Cyp27a1 maintained a similar circulating level of 25-hydroxyvitamin D3 and 1α,25-dihydroxyvitamin D3. Our results support the idea that the CYP2R1 is the major enzyme responsible for 25-hydroxylation of vitamin D, but clearly a second, as-yet unknown, enzyme is another contributor to this important step in vitamin D activation.

In this study, high-resolution (1)H NMR spectroscopy (600.03 MHz) and (13)C NMR spectroscopy (150.89 MHz) were used to elucidate the structures of equilibrating d-arabino-hexo-2-ulose (GLUC) (1) isomers in aqueous solution. Four isomers were formed from the investigated ketohexose, and their equilibrium is dependent on the pH value and temperature. Only hydrated GLUC (1) isomers were identified. The (2)C5-β-2,6-pyranoid and the β-2,5-furanoid GLUC (1) isomer were exclusively formed in aqueous solution. Thus, (4)C1-1,5-pyranoid isomers are predominating in the crystalline state. An increase in solution pH or temperature led to a pairwise conversion of configurative information. Thus, changing the measurement conditions permits control over the equilibrium's characteristic. Furthermore, all GLUC (1) isomers showed comparable reaction behavior regarding pH- and temperature-dependent degradation reactions.

Recent studies have shown that column stores can outperform row stores significantly. This paper explores alternative approaches to extend column stores with versioning, i.e., time travel queries and the maintenance of historic data. On the one hand, adding versioning can actually simplify the design of a column store because it provides a solution for the implementation of updates, traditionally a weak point in the design of column stores. On the other hand, implementing a versioned column store is challenging because it imposes a two dimensional clustering problem: should the data be clustered by row or by version? This paper devises the details of three memory layouts: clustering by row, clustering by version, and hybrid clustering. Performance experiments demonstrate that all three approaches outperform a (traditional) versioned row store. The efficiency of these three memory layouts depends on the query and update workload. Furthermore, the performance experiments analyze the time-space tradeoff that can be made in the implementation of versioned column stores.

[1] Ozone profiles in the upper mesosphere (70–100 km) retrieved from nine instruments are compared. Ozone from the Sounding of the Atmosphere using Broadband Emission Radiometry (SABER) instrument is used as the basis of comparison. Other measurements are from the Halogen Occultation Experiment, the High Resolution Doppler Imager, the Michelson Interferometer for Passive Atmospheric Sounding, the Global Ozone Monitoring by Occultation of Stars, the Atmospheric Chemistry Experiment—Fourier Transform Spectrometer, the Solar Occultation For Ice Experiment, the Optical Spectrograph and InfraRed Imaging System, and the Superconducting Submillimeter-Wave Limb-Emission Sounder. Comparisons of each data set with SABER using coincident profiles indicate agreement in the basic vertical profile of ozone but also some systematic differences in daytime ozone. Ozone from the SABER 9.6 μm channel is higher than the other measurements over the altitude range 60–80 km by 20–50%. Nighttime comparisons indicate better relative agreement (

Managing temporal data is becoming increasingly important for many applications. Several database systems already support the time dimension, but provide only few temporal operators, which also often exhibit poor performance characteristics. On the academic side, a large number of algorithms and data structures have been proposed, but they often address a subset of these temporal operators only. In this paper, we develop the Timeline Index as a novel, unified data structure that efficiently supports temporal operators such as temporal aggregation, time travel, and temporal joins. As the Timeline Index is independent of the physical order of the data, it provides flexibility in physical design; e.g., it supports any kind of compression scheme, which is crucial for main memory column stores. Our experiments show that the Timeline Index has predictable performance and beats state-of-the-art approaches significantly, sometimes by orders of magnitude.

We address the effect of an additional infrared radiative cooling/heating of the mesosphere and lower thermosphere (MLT) in the infrared bands of CO2, O3 and H2O due to small-scale irregular temperature fluctuations associated with gravity waves (GWs). These disturbances are not well resolved by present general circulation models (GCMs), but they alter the radiative transfer and cooling rates significantly. A statistical model of gravity wave-induced temperature variations was applied to large-scale temperature profiles, and the corresponding direct radiative calculations were performed with accounting for the breakdown of the local thermodynamic equilibrium (non-LTE). We show that temperature fluctuations can cause an additional cooling of up to 4 K day−1 near the mesopause. The effect is produced mainly by the fundamental 15 μm band of the main CO2 isotope 12C16O2 (626). A simple parametrization has been derived that computes corrections depending on the temperature fluctuations variance, which need to be added in radiative calculations to the mean temperature and the volume mixing ratios (VMRs) of CO2 and O(3P) to account for additional cooling/heating caused by the unresolved disturbances. Implementation of this scheme into the LIMA model resulted in a colder and broader simulated summer mesopause in agreement with recent lidar measurements at Spitsbergen.

The combination of satellite born SCIAMACHY hydroxyl and GOMOS ozone limb measurements allows for the derivation of the global distribution of atomic hydrogen abundance and instantaneous chemical heating rates in the mesopause region. Chemical heating rates show maximum values of 5–10 K/day at 85–90 km; atomic hydrogen densities are 1–5⋅108 cm−3. Signatures of equatorial Kelvin waves, Rossby-gravity waves and Rossby waves are clearly visible in the data. A pronounced latitudinal structure with maxima at the equator and at mid latitudes is observed. Between 2002 and 2008 chemical heating rates decreased and atomic hydrogen density increased, in accordance with model simulations of the 11-year solar cycle.

A 31-year–old white male patient was transferred from a local district hospital for suspected pericardial mass. Two months earlier he had been admitted there for worsening shortness of breath and subsequently was diagnosed with a large pericardial effusion. He had no other pertinent past medical history. After pericardiocentesis (1.5 L) there was no evidence of malignant cells on cytology, and a computed tomography scan did not reveal any abnormality. Thus, the incident was interpreted as (viral) infectious pericarditis and the patient clinically improved under anti-inflammatory medication within 4 weeks. After that, while on a holiday in Tunisia, he again reported worsening shortness of breath. On day 6 of his holiday, he suddenly passed out and was admitted to a local hospital for unexplained syncope, where again a large pericardial effusion was seen. This time, after another pericardiocentesis (1 L), a pericardial mass was suspected …

In this study, high-resolution 1H (600.03 MHz) and 13C NMR spectroscopy (150.89 MHz) were used to elucidate the structures of the identifiable isomers of a complex mixture of equilibrating 1-deoxy- d -erythro-hexo-2,3-diulose (1-DG) (1) isomers in aqueous solution. Approximately 15 isomers were formed from the investigated deoxy-ketohexose, and the equilibrium of the isomers is dependent on the pH values and temperature. In contrast to non-hydrated isomers, hydrated 1-DG (1) isomers were exclusively formed in aqueous solution. Their relative concentration increased when the pH was increased to more basic conditions. An increase in solution temperature led to the elimination of water and an increase in the relative concentration of the non-hydrated 1-DG (1) isomers. The observed influence of different conditions on the isomeric composition was explained for all isomers. Also, these observations provided information for structure elucidation. Furthermore, the reaction behaviour of 1-DG (1) under basic conditions at high temperatures (up to 347.5 K) was interpreted and described.

Fluidity and charging of supported bilayer lipid membranes (sBLMs) prepared from 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC) were studied by fluorescence recovery after photobleaching (FRAP) and microslit electrokinetic measurements at varying pH and ionic composition of the electrolyte. Measurements in neutral electrolytes (KCl, NaCl) revealed a strong correlation between the membrane fluidity and the membrane charging due to unsymmetrical water ion adsorption (OH(-) ≫ H(3)O(+)). The membrane fluidity significantly decreased below the isoelectric point of 3.9, suggesting a phase transition in the bilayer. The interactions of both chaotropic anions and strongly kosmotropic cations with the zwitterionic lipids were found to be related with nearly unhindered lipid mobility in the acidic pH range. While for the chaotropic anions the observed effect correlates with the increased negative net charge at low pH, no correlation was found between the changes in the membrane fluidity and charge in the presence of kosmotropic cations. We discuss the observed phenomena with respect to the interaction of the electrolyte ions with the lipid headgroup and the influence of this process on the headgroup orientation and hydration as well as on the lipid packaging.

Introduction: The ELN classification of cytogenetic aberrations in acute myeloid leukemia (AML) distinguishes favorable risk, intermediate risk I and II and adverserisk. The adverse-risk group contains patients (pts) with inv(3) and t(3;3). These pts have a significantly poorer outcome compared to other cytogenetic aberrations. The MRC classification considers both pts with inv(3) and t(3;3) as well as patients with other abn(3q) as adverse risk, but excludes t(3;5). Pts with inv(3) or t(3;3) have breakpoints located on the long arm of chromosome 3 at q21 and q26. As a result of these chromosomal modifications, an enhancer-protein is deregulated and the stem-cell regulator zinc finger protein EVI1 on 3q26 is over expressed. Other 3q aberrations do not involve EVI1. We conducted a comparative analysis on the impact of abn(3q) with likely EVI1 alteration versus abn(3q) without EVI1 involvement. Analyses were done both in the entire group of abn(3q) pts and in the subgroup of pts treated with allogeneic hematopoietic stem-cell transplantation (HSCT). Methods: We performed a retrospective analysis on 163 patients with an abnormality on the q arm of chromosome 3 (abn(3q)). These pts were treated between 1996 and 2009 in three multicenter studies by the German SAL study group (AML2003, AML96, AML60+). Pts with t(3;5) were excluded (n=11). The remaining 152 patients were divided into two groups. Group 1 (EVI1) contained 56 patients with a chromosomal aberration likely to alter EVI1, i.e. t(3;3), inv(3) and abn(3)(q26). Group 2 (noEVI1) comprised the remaining 96 patients displaying other abn(3q) aberrations. We compared groups for baseline characteristics, complete remission (CR), relapse-free survival (RFS) and overall survival (OS) in total and stratified for treatment. Results: Descriptive comparison of the groups (EVI1 vs noEVI1) revealed a significantly higher WBC count (14.3 vs 4.6 Gpt/l), PLT count (62 vs 47 Gpt/l) and -7 incidence (29% vs 16%) in the EVI1 group, whereas in the noEVI1 group, complex aberrations (25% vs 74%) and 17p alterations (0% vs 24%) occurred in a higher proportion of pts. CR rates (52% vs 47%), median RFS (7 vs 6 months) and median OS (6 vs 7 months) did not differ significantly between the two groups. In order to explore the clinical behavior of the different abn(3q) aberrations in relation to allogeneic HSCT, we compared EVI1 pts (n=21) versus noEVI1 pts (n=38) who received an allogeneic HSCT at any time during treatment. Patients with aberrant EVI1 were significantly younger (median age 44 vs 52 years), had a higher incidence of -7 (29% vs 13%), but less frequent karyotype complexity (10% vs 74%) or 17p alterations (0% vs 24%). More patients in the EVI1 group achieved a first CR before HSCT (95% vs 84%). Amongst CR pts, median RFS was slightly higher in the EVI1group (9 vs 6 months). In all abn(3q) pts with allogeneic HSCT, median OS was 30 months in the EVI1 group and only 12.5 months in the noEVI1 group. According to the log-rank test, this difference did not reach statistical significance (p=0.137). The advantage in mean OS for EVI1 patients is most likely due to the higher proportion of patients transplanted in CR while the accumulation of complex karyotypes in the noEVI1 group caused more primary resistant AML cases with a rapid progression even after allogeneic HSCT. Conclusions: Although AML development may be based on different molecularbiological mechanisms in patients with different abn(3q) aberrations depending on EVI1 alteration, the prognosis of the two groups is very similar. The most likely reason is the equal balance of favorable and adverse prognostic factors between the two groups such as age, karyotype complexity, 17p alteration and -7. Patients of both groups benefit from allogeneic HSCT to a similar extent. Confirmation of these results on larger data sets is desirable and under way. Disclosures Baldus: Novartis: Research Funding. Einsele:Novartis: Consultancy, Honoraria, Speakers Bureau; Amgen/Onyx: Consultancy, Honoraria, Speakers Bureau; Janssen: Consultancy, Honoraria, Research Funding, Speakers Bureau; Celgene: Consultancy, Honoraria, Research Funding, Speakers Bureau. Thiede:Novartis: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; AgenDix GmBH: Equity Ownership.

CYP24A1 is the cytochrome P450 component of the 25-hydroxyvitamin D(3)-24-hydroxylase enzyme that catalyzes the conversion of 25-hydroxyvitamin D(3) (25-OH-D(3)) and 1,25-dihydroxyvitamin D(3) (1,25-(OH)(2)D(3)) into 24-hydroxylated products, which constitute the degradation of the vitamin D molecule. This review focuses on recent data in the CYP24A1 field, including biochemical, physiological and clinical developments. Notable among these are: the first crystal structure for rat CYP24A1; mutagenesis studies which change the regioselectivity of the enzyme; and the finding that natural inactivating mutations of CYP24A1 cause the genetic disease idiopathic infantile hypercalcemia (IIH). The review also discusses the emerging correlation between rising serum phosphate/FGF-23 levels and increased CYP24A1 expression in chronic kidney disease, which in turn underlies accelerated degradation of both serum 25-OH-D(3) and 1,25-(OH)(2)D(3) in this condition. This review concludes by evaluating the potential clinical utility of blocking this enzyme with CYP24A1 inhibitors in various disease states.

To assess the optimal cumulative dose of cytarabine for treatment of young adults with acute myeloid leukemia (AML) within a prospective multicenter treatment trial.Between 1996 and 2003, 933 patients (median age, 47 years; range 15 to 60 years) with untreated AML were randomly assigned at diagnosis to receive cytarabine within the first consolidation therapy at either a intermediate-dose of 12 g/m² (I-MAC) or a high-dose of 36 g/m² (H-MAC) combined with mitoxantrone. Autologous hematopoietic stem-cell transplantation or intermediate-dose cytarabine (10 g/m²) were offered as second consolidation. Patients with a matched donor could receive an allogeneic transplantation in a risk-adapted manner.After double induction therapy including intermediate-dose cytarabine (10 g/m²), mitoxantrone, etoposide, and amsacrine, complete remission was achieved in 66% of patients. In the primary efficacy analysis population, a consolidation with either I-MAC or H-MAC did not result in significant differences in the 5-year overall (30% v 33%; P = .77) or disease-free survival (37% v 38%; P = .86) according to the intention-to-treat analysis. Besides a prolongation of neutropenia and higher transfusion demands in the H-MAC arm, rates of serious adverse events were comparable in the two groups.In young adults with AML receiving intermediate-dose cytarabine induction, intensification of the cytarabine dose beyond 12 g/m² within first consolidation did not improve treatment outcome.

SummaryIn the current work an attempt was made to apply the fundamental parameter method to the analysis of boron, oxygen, fluorine and heavier components of silica glasses. Experimental results are presented and compared with calculations based on theoretical work. The specimens consist of sets of certified standards which were obtained from Breitlahder company, Germany.Furthermore, it is shown that the match between the certified compositions and fundamental parameter computations is affected by the quality of available fundamental parameters for the ultralight elements as well as by the underlying mathematical models. Especially in quantitative analysis of these elements, a large variety of secondary enhancement effects must be taken into account. Two major mechanisms have been considered in the presented computations: excitation by fluorescence photons emitted from aiJ available excited atoms, and excitation by photoelectrons ejected after the primary ionization event. Their magnitude exceeds that of the conventional energy region by far and thus uncertainities in the corrective algorithms have a large effect

Streaming current measurements were performed on poly(N-isopropylacrylamid-co-carboxyacrylamid) (PNiPAAM-co-carboxyAAM) soft thin films over a broad range of pH and salt concentration (pH 2.5-10, 0.1-10 mM KCl) at a constant temperature of 22 °C. The films are negatively charged because of the ionization of the carboxylic acid groups in the repeat unit of the copolymer. For a given salt concentration, the absolute value of the streaming current exhibits an unconventional, nonmonotonous dependence on pH with the presence of a maximum at pH ∼6.4. This maximum is most pronounced at low electrolyte concentration and gradually disappears with increasing salinity. Complementary ellipsometry data further reveal that the average film thickness increases by a factor of ∼2.2 with increasing pH over the whole range of salt concentration examined. The larger the solution salt concentration, the lower the pH value where expansion of the hydrogel layer starts to take place. The dependence of film thickness on pH and electrolyte concentration remarkably follows that obtained for surface conductivity. The streaming current and surface conductivity results could be consistently interpreted on a quantitative basis using the theory we previously derived for the electrokinetics of charged diffuse (heterogeneous) soft thin films complemented here by the derivation of a general expression for the surface conductivity of such systems. In particular, the maximum in streaming current versus pH is unambiguously attributed to the presence of an interphasial gradient in polymer segment density following the heterogeneous expansion of the chains within the film upon swelling with increasing pH. A quantitative inspection of the data further suggests that pK values of ionogenic groups in the film as derived from the streaming current and surface conductivity data differ by ∼0.9 pH unit. Such a difference is attributed to the impact of position-dependent hydrophobicity across the film on the degree of ionization of carboxylic sites.

The authors report on the characterization of the charge formation at supported bilayer lipid membranes (sBLMs) prepared from the zwitterionic lipid 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine on planar silicon dioxide substrates. The charging of the sBLMs was studied in KCl solutions of different ionic strengths between 0.1 and 10 mM by streaming current measurements. In addition, attenuated total reflection Fourier transform infrared spectroscopy and fluorescence microscopy were applied to determine the lipid concentration in the membrane and to study the influence of the harsh conditions (pH 9-2, shear forces) during the electrokinetic measurements on the membrane stability and the lipid diffusion coefficient. The sBLMs were found to be extremely stable. Isoelectric points of about 4 revealed that unsymmetrical adsorption of hydroxide and hydronium ions determined the charging of the outer leaflet of the membrane in the investigated pH range. The diffusion coefficients were found to be rather independent on the ionic strength at neutral and alkaline pH. However, significantly decreased lipid diffusion at pH

The analysis of vitamins D and their metabolites and analogues has been reviewed by us on two occasions (Makin et al., 1995; Jones and Makin, 2000) over the last 10-15 years. In this chapter, we have drawn heavily on the 2000 review, up-dating it to take account of the developments in methodology that have occurred in the intervening years, but including elements of our 1995 review so that the reader can get a picture of the historical context as well as the modern developments.

The relative importance of different processes and transitions in the appearance of non-LTE populations for the fundamental levels of ozone was studied by quantifying the kinetic law of every process and transition that affect each level population. The vibrational temperatures and the relative contribution of every transition are presented as a function of altitude. The results show that the appearance of non-LTE populations for the fundamental levels is not produced by a direct imbalance between absorption and emission but as a consequence of imbalanced collisional transitions provoked by the overpopulation of O3(0 v2 0) levels. This fact confirms that the relaxation cascade drives through the bending mode ν2. © 2009 Elsevier Ltd. All rights reserved. Fil: Fernandez, Rafael Pedro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Kaufmann, Martin. Institute for Chemistry and Dynamics of the Geosphere; Alemania Fil: Toselli, Beatriz Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina

Contains fulltext : 79937.pdf (Publisher’s version ) (Closed access) From 2002 to 2007, 103 patients with primary myelofibrosis or postessential thrombocythemia and polycythemia vera myelofibrosis and a median age of 55 years (range, 32-68 years) were included in a prospective multicenter phase 2 trial to determine efficacy of a busulfan (10 mg/kg)/fludarabine (180 mg/m(2))-based reduced-intensity conditioning regimen followed by allogeneic stem cell transplantation from related (n = 33) or unrelated donors (n = 70). All but 2 patients (2%) showed leukocyte and platelet engraftment after a median of 18 and 22 days, respectively. Acute graft-versus-host disease grade 2 to 4 occurred in 27% and chronic graft-versus-host disease in 43% of the patients. Cumulative incidence of nonrelapse mortality at 1 year was 16% (95% confidence interval, 9%-23%) and significantly lower for patients with a completely matched donor (12% vs 38%; P = .003). The cumulative incidence of relapse at 3 years was 22% (95% confidence interval, 13%-31%) and was influenced by Lille risk profile (low, 14%; intermediate, 22%; and high, 34%; P = .02). The estimated 5-year event-free and overall survival was 51% and 67%, respectively. In a multivariate analysis, age older than 55 years (hazard ratio = 2.70; P = .02) and human leukocyte antigen-mismatched donor (hazard ratio = 3.04; P = .006) remained significant factors for survival. The study was registered at www.clinicaltrials.gov as #NCT 00599547.

XQuery is a declarative programming language which can be used to express queries and transformations of XML data. The goal of this paper is to explore the expressive power of XQuery as a general-purpose programming language. To this end, this paper describes how to build an entire enterprise web application in XQuery. It is shown that it is actually possible and quite effective to implement a web application entirely in XQuery and that there are several advantages in doing so. The resulting code has proven to be very concise and elegant. More importantly, the use of XQuery simplifies the overall application architecture and improves flexibility.

Studies of 25-hydroxyvitamin D 3 -24-hydroxylase (CYP24A1) have demonstrated that it is a bifunctional enzyme capable of the 24-hydroxylation of 1α,25-(OH) 2 D 3 , leading to the excretory form, calcitroic acid, and 23-hydroxylation, culminating in 1α,25-(OH) 2 D 3 -26,23-lactone. The degree to which CYP24A1 performs either 23- or 24-hydroxylation is species-dependent. In this paper, we show that the human enzyme that predominantly 24-hydroxylates its substrate differs from the opossum enzyme that 23-hydroxylates it at only a limited number of amino acid residues. Mutagenesis of the human form at a single substrate-binding residue (A326G) dramatically changes the regioselectivity of the enzyme from a 24-hydroxylase to a 23-hydroxylase, whereas other modifications have no effect. Ala-326 is located in the I-helix, close to the terminus of the docked 25-hydroxylated side chain in a CYP24A1 homology model, a result that we interpret indicates that substitution of a glycine at 326 provides extra space for the side chain of the substrate to move deeper into the pocket and place it in a optimal stereochemical position for 23-hydroxylation. We discuss the physiological ramifications of these results for species possessing the A326G substitution, as well as implications for optimal vitamin D analog design.

[1] Nonlocal thermodynamic equilibrium (non-LTE) simulations of the (CO)-C-12-O-16(1 -> 0) fundamental band, the (CO)-C-12-O-16(2 -> 1) hot band, and the isotopic (CO)-C-13-O-16(1 -> 0) band performed with the Generic Radiative Transfer and non-LTE population Algorithm (GRANADA) and the Karlsruhe Optimized and Precise Radiative Transfer Algorithm (KOPRA) have been compared to spectrally resolved 4.7 mu m radiances measured by the Michelson Interferometer for Passive Atmospheric Sounding (MIPAS). The performance of the non-LTE simulation has been assessed in terms of band radiance ratios in order to avoid a compensation of possible non-LTE model errors by retrieval errors in the CO abundances inferred from MIPAS data with the same non-LTE algorithms. The agreement with the measurements is within 5% for the fundamental band and within 10% for the hot band. Simulated (CO)-C-13-O-16 radiances agree with the measurements within the instrumental noise error. Solar reflectance at the surface or clouds has been identified as an important additional excitation mechanism for the CO( 2) state. The study represents a thorough validation of the non-LTE scheme used in the retrieval of CO abundances from MIPAS data.

A systematic analysis of conserved H-bonding patterns and tertiary structural motifs from 13 crystal structures was used to create a homology model for the human multicatalytic cytochrome P450, CYP24A1, involved in catabolism of 1alpha,25-dihydroxyvitamin D3. The substrate was docked in the active site and used to identify potential substrate contact residues in the B' helix, B'/C loop, F-helix and the beta-5 hairpin. Seven CYP24A1 mutants were created and studied by mammalian cell transfection and CYP24A1 activity assay. Mutants showed reduced metabolic rates and altered metabolite patterns compared to wild-type. We conclude that: Ile-131 positions substrate via A-ring and cis-triene contacts; Trp-134 and Gly-499 are determinants of substrate access; Leu-148 contacts the substrate side-chain; Met-246 is important in mediating regioselectivity. Our findings validate the new model of CYP24A1, which can now be used to predict structural modifications for rational vitamin D drug design.

Like most pharmaceutical agents, vitamin D analogs are subject to hepatic metabolism by a variety of cytochrome P450 (CYP)-based systems. Metabolism can involve activation as well as inactivation of the vitamin D analog and one of the more successful families includes the 1alpha-hydroxyvitamin D prodrugs (1alpha-OH-D2, 1alpha-OH-D3, 1alpha-OH-D4, 1alpha-OH-D5), that all require a step of activation. Some of these prodrugs are in use or clinical trial because they have a therapeutic advantage over calcitriol. However, the nature of the activation of these molecules is poorly understood, particularly with regard to the CYP isoform involved. Various transfected CYPs and hepatic cell lines combined with tandem LC-MS analysis were used to investigate the metabolism of a spectrum of vitamin D analogs, including 1alpha-OH-Ds and the topical analog, calcipotriol. In the case of the 1alpha-OH-Ds, evidence was found of multiple sites of side-chain hydroxylation consistent with the generation of more than one active form. The potential involvement of CYP27A and other putative 25-hydroxylases in 1alpha-OH-D activation was also shown, as well as the potential for CYP24 activation and inactivation. In the case of calcipotriol, the respective roles of non-vitamin D-related CYPs and CYP24 in the catabolism of this anti-psoriatic drug were dissected out using cell lines with or without CYP24 expression, allowing us to demonstrate the potential contribution of CYP24 to "vitamin D resistance". The implications of hepatic metabolism in the context of other facets thought to play a role in the mechanism of action of anticancer and antiproliferative vitamin D analogs are discussed.

Introduction: Sequential high dose (SHiDo) chemotherapy with stem cell support has been shown to prolong the event-free survival in patients with diffuse large B-cell lymphoma. Methods: To confirm this result in a multicenter trial, we randomized patients with aggressive NHL, to receive either eight cycles of CHOP or SHiDo. The primary endpoint was overall survival. Results: 129 evaluable patients were randomized to receive either CHOP or SHiDo: median age, 48 years; 62% male; stage III+IV: 73%; age adjusted International Prognostic Index 1/2/3: 21%/52%/27%. Toxicity grades 3+4 were more pronounced in the SHiDo-arm with 13% versus 3% of patients with fever; 34% versus 13% with infections; 13% versus 2% with esophagitis/dysphagia/gastric ulcer. The remission rates were similar in SHiDo and CHOP arms with 34%/37% complete remissions and 31%/31% partial remissions, respectively. After a median observation time of 48 months, there was no difference in overall survival at 3 years, with 46% for SHiDo and 53% for CHOP ( P= 0.48). Conclusion: In this multicenter trial, early intensification with SHiDo did not confer any survival benefit in previously untreated patients with aggressive NHL and was associated with a higher incidence of grades 3/4 toxicity.

The best strategy for incorporating imatinib in front-line treatment of Ph+ acute lymphoblastic leukemia (ALL) has not been established. We enrolled 92 patients with newly diagnosed Ph+ ALL in a prospective, multicenter study to investigate sequentially 2 treatment schedules with imatinib administered concurrent to or alternating with a uniform induction and consolidation regimen. Coadministration of imatinib and induction cycle 2 (INDII) resulted in a complete remission (CR) rate of 95% and polymerase chain reaction (PCR) negativity for BCR-ABL in 52% of patients, compared with 19% in patients in the alternating treatment cohort (P = .01). Remarkably, patients with and without a CR after induction cycle 1 (INDI) had similar hematologic and molecular responses after concurrent imatinib and INDII. In the concurrent cohort, grades III and IV cytopenias and transient hepatotoxicity necessitated interruption of induction in 87% and 53% of patients, respectively; however, duration of induction was not prolonged when compared with patients receiving chemotherapy alone. No imatinib-related severe hematologic or nonhematologic toxicities were noted with the alternating schedule. In each cohort, 77% of patients underwent allogeneic stem cell transplantation (SCT) in first CR (CR1). Both schedules of imatinib have acceptable toxicity and facilitate SCT in CR1 in the majority of patients, but concurrent administration of imatinib and chemotherapy has greater antileukemic efficacy.

Introduction Aberrant DNA methylation is a common feature of acute myeloid leukemia (AML) and increases with age. DNMT inhibitors such as Azacitidine (AZA) can induce meaningful responses and remissions in AML as monotherapy. The combination of AZA with standard chemotherapy (7+3) has not been tested in a randomized trial. Patients and study design The AML-AZA trial compared AZA directly followed by standard induction therapy, AZA followed by standard consolidation, and further Azacitdine maintenance with standard induction and consolidation without AZA in older patients with AML. All patients received standard Cytarabine (100 mg/sqm) and Daunorubicin (60 mg/sqm) induction (“7+3”) and up to two cycles of intermediate dose Cytarabine (1 g/sqm q12hr days 1, 3, 5) as consolidation therapy. AZA (75 mg/sqm for 5 days) preceded each therapy cycle in the AZA arm. In addition, AZA maintenance for up to 1 year was also scheduled for patients in the AZA arm. 105 patients were randomized to receive AZA plus Cytarabine plus Daunorubicin as induction therapy (AZA + 7+3) and 109 patients to receive 7+3 only (control group). Median age was 70 years in both treatment arms. Patient cohorts were well balanced with regard to blast counts in bone marrow, secondary versus de novo AML and molecular genetics risk group. More patients in the AZA + 7+3 arm (39/105; 37.1%) than in the control group (25/109; 22.9%) showed high risk cytogenetics (p=0.057). Event free survival (EFS) was the primary end point. Secondary endpoints were overall survival (OS), complete remission (CR) rate, toxicity and different treatment response according to molecular markers. Results Overall, 214 of 216 planned patients were enrolled into the AML-AZA trial. Due to a higher number of severe adverse events (SAE), AZA administration was stopped after recruitment of 214 patients whereas chemotherapy was continued as planned. Percentages of patients in the AZA arm with AZA doses as initially planned were as follows: 99% for first induction cycle, 72% for the second induction cycle. AZA as maintenance therapy for at least one cycle was delivered to 18% of patients in the AZA group. At least one SAE occurred in 51% of AZA + 7+3 patients compared to 31% of 7+3 patients (p=0.005). Cardiac disorders with CTCAE grade 3-5 occurred more frequently in the AZA + 7+3 arm (n = 15) than in the 7+3 arm (n = 6) (not significant). Leukopenia was prolonged by one day (median 23 vs 22 days) in the AZA + 7+3 group (p=0.043), whereas time of thrombocytopenia was not different. The early death rates at 30, 60 and 90 days did not differ significantly between treatment groups. Efficacy analyses were performed on an intention-to-treat basis. Median EFS as the primary endpoint was 6 months in both treatment arms (p=0.96). Median OS was 16 months for patients treated with AZA + 7+3 and 21 months for 7+3 (p=0.35). Median relapse free survival was 12 months in both treatment arms (p=0.95). 48 of 100 patients (48%) in the AZA + 7+3 arm achieved complete remission (CR) after induction therapy versus 57 of 109 patients (52%) in the 7+3 arm (p=0.58). DNMT3A exon 23 mutations were detected in 30 out of 162 analyzed patients. Exploratory analyses were performed to detect a potential interaction between AZA + 7+3 response and DNMT3A mutation status. Trends for improved EFS and OS were noted for AZA + 7+3 treatment in DNMT3A mutated patients. Conclusion AZA as addition prior to standard induction and consolidation chemotherapy does not prolong EFS and OS in unselected older AML patients and it is more toxic. However, a trend towards better efficacy in patients with DNMT3A mutation was observed and should be further explored. Disclosures Müller-Tidow: Celgene: Honoraria, Research Funding. Thiede:AgenDix GmbH: Equity Ownership, Research Funding; Illumina: Research Support, Research Support Other. Kiehl:Roche: Membership on an entity's Board of Directors or advisory committees. Brümmendorf:Novartis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties, Research Funding. Ehninger:GEMoaB GmbH: Consultancy, Patents & Royalties.

Studies to elucidate the role of dehydroepiandrosterone (DHEA) metabolism in neuroprotection have compared its relative 7-hydroxylation against estrogen formation by way of 4-androstenedione (AD) in various rodent brain cell lines. In all cases, the 7alpha- and 7beta-hydroxy epimers of DHEA were found to be the dominant products with one notable exception. BV2 mouse microglia were virtually unable to hydroxylate DHEA at C-7 and converted AD to a major unknown metabolite not observed with mouse BHc hippocampal cells. In this paper, we describe the identification of this compound based on its physical properties and analysis by TLC and HPLC. Its identity as 3beta-hydroxy-4-androstene-17-one, the Delta(4)-isomer of DHEA, was confirmed by mass spectrometry (LC/MS), as well as by reverse isotope dilution analysis involving co-crystallization with the synthetic steroid. Possible mechanisms for the formation of this isomer of DHEA by BV2 microglia are proposed, together with that of other C-19 steroids detected which include testosterone (T), 5alpha-dihydrotestosterone and 5alpha-androstanedione.

Feacal samples from 630 slaughtered healthy pigs were examined with the aim: (i) to monitor the shedding of non-O157 STEC and E. coli O157; (ii) to further characterize isolated strains; and (iii) to discuss the results obtained with their relevance to food safety. The percentage of the 630 samples testing positive for STEC by polymerase chain reaction was 22.2%. The 32 isolated stx positive strains (31 sorbitol-positive) belonged to non-O157 STEC and comprised ten serotypes. Stx1, stx2, and both toxin genes were detected in 3%, 97%, and 0% of strains. Among stx2-positive strains, 29 were positive for stx2e, one for stx2/stx2e, and one for stx2c/stx2e. The percentage of the 630 samples testing positive for E. coli O157 by polymerase chain reaction was 7.5%. The 31 isolated rfbE PCR positive strains tested negative for stx genes. However, four strains harbored eae and were classified as EPEC. Introduction The importance of the Shiga toxin (Stx) -producing Escherichia coli (STEC) group has increased since a food-borne infection caused by STEC has first been reported in 1983 (Riley et al, 1983). Pathogenicity of STEC in humans is associated with diverse virulence factors. The main factors are the ability to form cytotoxic exotoxins (shiga toxins), which can be subdivided into a shiga toxin 1 group (Stx1 group) and a shiga toxin 2 group (Stx2 group), as well as the property of producing attachment-effacement lesions and the presence of an enterohaemolysin gene (EHEC- hlyA) (Boerlin et al, 1999). The source of STEC food-borne infection was often found to be foods of bovine origin or other fecally cross-contaminated foods. Cattle are currently considered the main reservoir of STEC path- ogenic to humans. The results of a Swiss study of faeces of healthy beef cattle at slaughter showed a STEC carriage rate ranging from 2.3% to 23.7% (Stephan et al, 2000). In addition, there are some reports on the isolation of Stx positive E.coli from pigs, but often strains originating from pigs with diarrhoea or oedema disease were tested (Moon et al, 1999; Osek 1999). Although E. coli of the serogroups O138, O139 and O141 producing the Stx2e variant can cause oedema disease in pigs, they seem not to be able to colonized the human intestinal tract. However, since strains harbouring stx2e have also been isolated from patients with diarrhea

[1] The Michelson Interferometer for Passive Atmospheric Sounding (MIPAS) onboard the Envisat satellite provides temperature and various gas profiles from limb-viewing midinfrared emission measurements. The stratospheric temperatures retrieved at the Institut fur Meteorologie und Klimaforschung (IMK) for September/October 2002 and October/November 2003 are compared with a number of reference data sets, including global radiosonde (RS) observations, radio occultation (RO) measurements of Global Positioning System (GPS) on German Challenging Minisatellite Payload ( CHAMP) and Argentinean Satelite de Aplicaciones Cientificas-C (SAC-C) satellite, Halogen Occultation Experiment (HALOE) on the Upper Atmosphere Research Satellite (UARS), and the analyses of European Centre for Medium-Range Weather Forecasts (ECMWF) and Met Office (METO), United Kingdom. The data sets show a good general agreement. Between 10 and 30 km altitude the mean differences are within +/- 0.5 K for the averages over the height interval and within +/-( 1 - 1.5) K at individual levels for comparisons with RS, GPS-RO/CHAMP, and SAC-C, ECMWF, and METO. Between 30 and 45 km the MIPAS mean temperatures, averaged over the height region, are higher than ECMWF but lower than METO by ∼ 1.5 K, while they differ by +/- 0.5 K with respect to HALOE, with maximum discrepancies of ∼ 2.5 K peaking around 35 km. Between 45 and 50 km, MIPAS temperatures show a low bias compared to HALOE, ECMWF, and METO with mean differences of - 1 to - 3 K and with a better agreement with HALOE. The large discrepancies between MIPAS and the analyses above 30 km likely suggest deficiency in the underlying general circulation models. The standard deviations vary between 2.5 and 3.5 K for individual data sets, with more than 70% being contributed from the expected variability of the atmosphere. Retrieved temperatures with accuracy of ∼ 0.5 - 1 K after removing the atmospheric variability provide highly accurate knowledge to characterize our environment.

Background: Vitamin D may be important in the causal pathway to breast cancer (BC) by influencing breast density, a potential intermediate biomarker for carcinogenesis. However, previous study results are inconsistent, particularly among postmenopausal women. Our objectives were to prospectively examine the relationship between blood biomarkers of vitamin D (serum 25-OH-D) and mammographic breast density (MD) in postmenopausal women at northern latitudes. Potential effect modifications by calcium or exemestane therapy on the relationship between serum 25-OH-D and follow-up MD were also examined. Methods: The NCIC Clinical Trials Group conducted a randomized controlled trial (RCT) comparing exemestane (E) with placebo (P) in 4560 postmenopausal women at high risk for developing BC (MAP.3) (Goss, NEJM 2011). Using a nested observational design, we evaluated a cohort of participants from across Canada and Buffalo, NY. BC risk factor information was previously collected; primary exposure and outcome data were obtained from serum samples and mammograms. Serum 25-OH-D was measured using state-of-the-art LC-MS/MS from samples collected at baseline and year 1. Values for year 1 samples were adjusted to match the month of sampling for the baseline sample month and these samples were averaged. Baseline and ≥ 3 year follow-up percent MD were centrally assessed, blinded to treatment allocation and vitamin D levels, using Cumulus software. Linear regression was used to estimate the effect of serum 25-OH-D on log transformed percent MD at ≥ 3 year follow-up controlling for month of sampling and potential confounders of age, body mass index, first degree family history of BC and calcium. Results: Percent MD was measured for 568 participants (E=287, P=281) with a ≥ 3 year follow-up mammogram obtained from 21 (91%) Canadian sites participating in MAP.3 and one US site in Buffalo, NY. The mean age of these women at study entry was 62 years and they were followed for a mean of 3.7 years. The geometric mean percent MD of the follow-up mammograms was 4.3%. A random sample of 10% (N=102) of mammograms was repeated with high intra-rater reliability (correlation coefficient = 0.95). Unadjusted for month of blood collection, the mean serum 25-OH-D concentration was 36.5 ng/mL (SD=10.6) based on pooled baseline and year one serum samples. After controlling for month of sampling and potential confounders, serum 25-OH-D was not predictive of log transformed percent MD (p=0.25). Statistically significant interactions with calcium (above vs. below the median calcium value p=0.30) or exemestane (p=0.99) on the serum 25-OH-D and MD relationship were not detected. Preliminary results suggest some evidence that low vitamin D ( Conclusion: In this nested observational study, no association between serum 25-OH-D levels and percent MD at ≥ 3 year follow-up was detected after controlling for month of sampling and many well established BC risk factors. This study was funded by the Canadian Breast Cancer Foundation. The MAP.3 trial was supported by Pfizer. The NCIC CTG is supported by the Canadian Cancer Society Research Institute. Citation Format: Melanie Walker, Harriet Richardson, Paul Goss, Doris Jabs, Glenville Jones, Martin Kaufmann, Jean Wactawski-Wende, Angela Cheung, Eric Winquist, Silvana Spadafora, Susan Ellard, Amanda Hey, Andrew Cooke, Andrea Eisen, Shailendra Verma, Lavina Lickley, Dongsheng Tu, Karen Gelmon, Ralph Meyer, Will King. Vitamin D and mammographic density in postmenopausal women: A cohort study nested within the NCIC CTG MAP.3 chemoprevention trial. [abstract]. In: Proceedings of the Eleventh Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2012 Oct 16-19; Anaheim, CA. Philadelphia (PA): AACR; Cancer Prev Res 2012;5(11 Suppl):Abstract nr B97.

Background Patients with follicular (FL) or mantle cell lymphoma (MCL) are incurable with conventional therapy. We investigated the safety and efficacy of rituximab consolidation after autologous stem cell transplantation (ASCT) in order to prevent relapse by clearance of minimal residual disease (MRD). Methods Rituximab was given ∼8 weeks after CD34+ cell enriched ASCT at 375 mg/m2, weekly for 4 weeks. Monitoring of MRD was performed by repetitive PCR analyses. Results Thirty-one patients were included; one died early after ASCT before rituximab administration. Thirty patients (20 FL, 10 MCL) were evaluable after rituximab consolidation, and 27 of these were assessable for MRD detection. Rituximab consolidation post-ASCT was safe, the most common toxicity being infection. At a median follow-up of 42 months (range 13–96) after ASCT, 25 patients were censored with an actuarial event-free survival (EFS) of 81% at 4 and 5 years. Four patients (two FL, two MCL) relapsed, and one additional MCL patient died unexpectedly in complete remission. PCR-negativity was observed in 22% of the patients before ASCT, 53% post-ASCT (P=0.0547), 72% after rituximab (P=0.0018) and 100% at 6 months post-transplant (P Conclusions One single course of rituximab consolidation given after ASCT is safe, may help to eliminate MRD and may translate into improved EFS in both FL and MCL patients.

The development of novel gene expression systems for cytochrome P450s (CYPs) together with a revolution in analytical mass spectrometry with the emergence of liquid chromatography/mass spectrometry (LC/MS) has opened the door to answering some long-standing questions in Vitamin D metabolism. Our studies focused on: (1) elucidating the role of CYP24 in 25-OH-D 3 and 1α,25-(OH) 2 D 3 metabolism; (2) exploring how DBP influences this process; (3) measuring 25-OH-D 3 metabolism in CYP24–knockout (CYP24–XO) cells and; (4) comparing 1α-OH-D 2 metabolism in the CYP24-XO mouse in vivo and in vitro. Methodology employed CYP24 over-expression and knockout systems in conjunction with state-of-the-art analytical LC/MS, diode array, and radioisotopic detection methods. We found that CYP24 metabolizes 25-OH-D 3 and 1α,25-(OH) 2 D 3 at similar rates in vitro, but that for 25-OH-D 3 but not 1α,25-(OH) 2 D 3 , this rate is strongly influenced by the concentration of DBP. Unlike their wild type littermates, the administration of 25-OH-D 3 to CYP24-XO mice results in no measurable 24,25-(OH) 2 D 3 production. When neonatal murine keratinocytes are prepared from wild type and CYP24-XO mice there was no measurable production of 24,25-(OH) 2 D 3 or 1α,24,25-(OH) 2 D 3 in CYP24-XO mice. Similar experiments using the same wild type and CYP24-XO animals and cells and [ 3 H ] 1α-OH-D 2 resulted in the apparent paradox that the Vitamin D prodrug was 25-hydroxylated in vivo but 24-hydroxylated in vitro.

The elucidation of the metabolic pathway for vitamin D, including the delineation of the specific cytochrome P450s (CYPs) involved in activation and catabolism, has emphasized the overall importance of metabolic considerations in vitamin D analog design. This short review attempts to summarize recent findings with isolated CYPs and animal models in which CYPs are genetically manipulated to draw attention to structural features of vitamin D analogs that make them more or less resistant to metabolic enzymes. We conclude by placing metabolic considerations in the context of the other important aspects of vitamin D analogs.

[1] The global distribution of mesospheric and lower thermospheric ozone 9.6 μm infrared emissions was measured by the Cryogenic Infrared Spectrometers and Telescopes for the Atmosphere (CRISTA) experiment during two Space Shuttle missions in November 1994 and August 1997. The radiances measured by CRISTA have been inverted to O3 number densities in the 50–95 km range by using a nonlocal thermodynamic equilibrium model. A detailed sensitivity study of retrieved O3 number densities has been carried out. The ozone abundance profiles show volume mixing ratios of 1–2 ppmv at the stratopause, 0.5 ppmv or less around 80 km, and typically 1 ppmv during daytime and 10 ppmv during nighttime at the secondary maximum. The agreement with other experiments is typically better than 25%. The global distribution of upper mesospheric ozone shows significant latitudinal gradients and an enhancement in the equatorial upper mesosphere. At the polar night terminator a third ozone maximum is observed. Three-dimensional model results indicate that the latitudinal gradients are significantly influenced by solar tides.

The elucidation of the metabolic pathway for vitamin D, including the delineation of the specific cytochrome P450s (CYPs) involved in activation and catabolism,has emphasized the overall importance of metabolic considerations in vitamin D analog design. This short review attempts to summarize recent findings with isolated CYPs and animal models in which CYPs are genetically manipulated to draw attention to structural features of vitamin D analogs that make them more or less resistant to metabolic enzymes. We conclude by placing metabolic considerations in the context of the other important aspects of vitamin D analogs.

During its two missions of about one week duration each in November 1994 and in August 1997 the Cryogenic Infrared Spectrometers and Telescopes for the Atmosphere (CRISTA) experiment observed spectrally resolved mid- and far- infrared emissions from the earth limb in the altitude regime from the troposphere to the lower thermosphere. The measurements concentrated on high spatial resolution in all three dimensions in the lower part of the height range. At upper mesosphere/lower thermosphere altitudes global ozone concentrations during day and night, daytime carbon-dioxide densities, and atmospheric temperatures were derived using a comprehensive non-LTE model. The derived parameters compare well with other experimental data. Pronounced horizontal structures in latitude as well as in longitude are found in all parameters showing that dynamical influences are of importance also in the upper mesosphere/lower thermosphere. Carbon dioxide mixing ratios start to depart from their tropospheric values at altitudes as low as 70 - 80 km. Atmospheric temperatures of 140 K and below were retrieved near the mesopause at high northern latitudes in August. These data are supported by the simultaneous observation of polar mesospheric clouds.

Bacterial pathogens require iron for proliferation and pathogenesis. Pseudomonas aeruginosa is a prevalent Gram-negative opportunistic human pathogen that takes advantage of immunocompromised hosts, and encodes a number of proteins for uptake and utilization of iron. Here we report the crystal structures of PhuS, previously known as the cytoplasmic heme-trafficking protein from P. aeruginosa, in both the apo- and holo-forms. In comparison to its homologue ChuS from Escherichia coli O157:H7, the heme orientation is rotated 1800across the α-γ axis, which may account for some of the unique functional properties of PhuS. In contrast to previous findings, heme binding does not result in an overall conformational change of PhuS. We employed spectroscopic analysis and CO measurement by gas chromatography to analyze heme degradation, demonstrating that PhuS is capable of degrading heme using ascorbic acid or cytochrome P450 reductase-NADPH as an electron donor, and produces five times more CO than ChuS. Addition of catalase slows down, but does not stop PhuS-catalyzed heme degradation. Through spectroscopic and mass spectrometry analysis, we identified the enzymatic product of heme degradation to be verdoheme. These data taken together suggest that PhuS is a potent heme-degrading enzyme, in addition to its proposed heme-trafficking function.

Purpose The treatment success in patients (pts) with acute myeloid leukemia (AML) is very heterogeneous. Cytogenetic and molecular alterations present at diagnosis are strong prognostic factors, which have been used to individualize treatment. As shown by several groups, the subgroup of pts with deletion of the short arm of chromosome 17 are at high risk for treatment failure (e.g. Seifert, Leukemia 2009), which persists even after allogeneic hematopoietic stem cell transplantation (HSCT) (Middeke, Blood 2012; Mohr, Br J Haematol 2013). Besides allelic loss of TP53 located on the short arm of chromosome 17, other mechanisms of inactivation have been shown for this key tumor suppressor gene, most importantly missense point mutations or small deletions. These alterations have also been linked to poor outcome in AML after chemotherapy (Grossmann, Blood 2012). Here, we studied the impact of TP53 mutations on the outcome of AML pts with adverse cytogenetic risk treated with HSCT. Patients and Methods We selected AML pts with complex karyotype (CK), monosomy 7, monosomy 5/del5q and/or abnl(17p) who had received HSCT within 3 randomized controlled trials (NCT numbers 00180115, 00180102, and 00180167). All pts were treated with intensive induction chemotherapy and HSCT according to a risk adapted strategy. Complete sequencing of the TP53coding region was done using next generation sequencing (NGS) on a 454 GS Junior instrument (Roche) using the IRONII-study amplicon panel. Amplicons were generated from genomic DNA isolated at the time of diagnosis. Data analysis was done using the Sequence Pilot software package (JSI Medical Systems), a 10% cut-off was used for mutation calling. Nonsynonymous mutations were classified into bi-allelic TP53 mutations if detected allelic frequency as determined by NGS was >50% and mono-allelic TP53 mutations for frequencies between 10% and 50%. All samples with synonymous mutations or no detectable mutations according to the predefined cut-off of 10% were classified as TP53wild type (wt). Overall survival (OS), event-free survival (EFS), cumulative incidence of relapse (CIR) and non-relapse-mortality (NRM) after HSCT were analyzed according to the mutational status. Results Samples from 97 pts with AML were analysed, the median age was 51 years (range 18 to 67), 83% suffered from de novo AML, while 13% had sAML and 3% therapy-related myeloid neoplasms. CK and monosomal karyotype (MK) were present in 61% and 42% of the pts, respectively. Twenty-nine pts (30%) had abnl(17p) detected by conventional karyotyping or FISH analysis. Twenty-six pts (27%) were treated with standard myeloablative conditioning (MAC) regimens while the remaining pts received reduced intensity conditioning (RIC). Donors were siblings in 36 pts (37%) and matched or mismatched unrelated donors in all other pts. Overall, TP53 mutations were found in 40 pts (41%). Twenty-eight (29%) pts had a bi-allelic TP53 mutation while 12 (12%) pts had a mono-allelic TP53 mutation. We identified 15 pts with TP53 mutations without abnl(17p). Four pts with abnl(17p) had wt TP53. Pts with TP53 mutations were significantly older than pts with wt TP53 AML (median age 55 vs. 43, p=.004). Donor type, type of conditioning and the rate of transplantation in first complete remission were not statistically different among pts with or without TP53mutations. With a median follow up of 67 months the three-year probabilities of OS and EFS for pts with wt TP53 were 33% (95% CI, 21% to 45%) and 24% (95% CI, 13% to 35%) compared to 10% (95% CI, 0% to 19%) and 8% (95% CI, 0% to 16%) (p=.002 and p=.007) for those with mutated TP53, respectively. CIR at three years was 42% for pts with wt TP53 and 60% for those with mutated TP53 (p=.05). NRM was not different in both groups. In multivariate analysis including age, donor type (sibling vs. all other), type of conditioning (RIC vs. MAC) and disease status (CR1 vs. advanced disease) only the TP53-mutation status had a significant influence on EFS (HR=1.72; p=.03). In our analysis, classification according to MK did not significantly influence OS, EFS, CIR or NRM. Conclusion In this cohort of pts with cytogenetic adverse risk abnormalities, who had received HSCT, TP53 mutations were present in 41% of the pts. OS and EFS were significantly worse in pts with mutated TP53. Mutational analysis of TP53 might be an important additional tool to predict outcome after HSCT in pts with adverse karyotype AML. Disclosures: No relevant conflicts of interest to declare.

Abstract 474 BACKGROUND: Autologous stem cell transplantation (autoSCT) is considered as standard treatment for non-frail patients with mantle cell lymphoma (MCL). However, little is known about outcome of MCL recurrence after autoSCT. We therefore conducted a retrospective analysis of patients with MCL who failed autoSCT using the EBMT database. PRIMARY OBJECTIVE was to analyse outcome and prognostic factors after relapse following autoSCT for MCL in the rituximab era. PRIMARY ENDPOINT was overall survival (OS) from relapse. ELIGIBLE were patients aged 18 years or more who relapsed following an autoSCT for MCL performed between 2000 and 2010 and who were registered with the EBMT. Centres were contacted to provide additional information on relapse treatment. STATISTICAL ANALYSIS was based on log-rank comparisons and multivariable testing using Cox regression models. RESULTS: 1054 patients meeting the eligibility criteria could be identified in the EBMT registry. Of these, a full data set could be retrieved for 382 patients. Sixteen patients had to be excluded due to loss of follow up (n=7), wrong diagnosis (n=6), or falsely reported relapse (n=3). Median age at autoSCT of 366 evaluable patients was 59 years (range: 37 to 76), 290 patients (79%) were men. 64% had undergone autoSCT as part of 1st-line therapy; 68% and 49% had documented exposure to rituximab (RTX) and high-dose ara-C (HA) before autoSCT; and 12% had had refractory disease at autoSCT. Median time from autoSCT to relapse was 20 months (range: 0.4 to 117). 21 relapses (6%) occurred beyond 5 years after autoSCT. With a median observation time of 37 months (95% CI 32–43), median OS after relapse of the whole study group was 20 months. By univariate analysis, a long (>12mo) interval between autoSCT and relapse (p Salvage chemotherapy after relapse resulted in only 31% complete responses and 29% partial responses, whereas 40% of patients have been refractory to first salvage chemotherapy. 83 patients (23%) received an allogeneic SCT (alloSCT), whereas only 7 patients (2%) received a second autoSCT after relapse. Median time after relapse to second SCT was 7 months (range: 1 to 40). Survival after relapse for patients who received a second autoSCT was poor with no long-term survivor. AlloSCT performed for late relapse (>12mo) after autoSCT was associated with superior OS compared to patients who received an allograft upon a shorter remission duration after autoSCT (5-year OS from alloSCT 50% vs 0%; p=0.001; Figure 1B). Achievement of CR before alloSCT (p=0.05 HR=0.5), but not donor source, T-cell depletion or conditioning intensity affected OS after alloSCT. CONCLUSIONS: Patients with MCL who relapse within one year after autoSCT have an extremely dismal outcome even with alloSCT. In contrast, about half of the patients who have MCL recurrence beyond one year after autoSCT and can undergo salvage alloSCT enjoy long-term survival. It remains to be shown if a similarly good outcome can be achieved without alloSCT in this favourable selection of patients. A 2nd autoSCT does not appear to be a promising option in patients with MCL failing a 1st autoSCT. Disclosures: Walewski: Mundipharma: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Roche: Honoraria, Speakers Bureau; GSK: Membership on an entity's Board of Directors or advisory committees; Celgene: Membership on an entity's Board of Directors or advisory committees; Janssen-Cilag: Membership on an entity's Board of Directors or advisory committees; Cephalon: Research Funding.

Abstract 332 PURPOSE : In Multiple Myeloma (MM), the combination of serum beta-2-microglobulin level with serum albumin concentration has been proposed as an outcome predictor in the International Staging System (ISS). More recently, subgroups of MM defined by genetic and cytogenetic abnormalities have been associated with unique biologic, clinical, and prognostic features. PATIENTS AND METHODS: We analyzed the prognostic value of 12 chromosomal abnormalities by fluorescent in situ hybridization (FISH) in a series of 354 MM patients treated within the HOVON-65/GMMG-HD4 trial. Patients with newly diagnosed MM were randomized to receive either three cycles of VAD (arm A; vincristine, adriamycin, dexamethasone) or PAD (arm B; bortezomib, adriamycin, dexamethasone). All patients underwent autologous stem cell transplantation (ASCT) followed by maintenance therapy with thalidomide 50 mg daily (arm A) or bortezomib 1.3 mg/m2 once every 2 weeks (arm B), respectively. In addition, a second cohort of patients was analyzed as a control group (n=462), undergoing ASCT at the University of Heidelberg between September 1994 and December 2010. RESULTS: For the entire group of patients treated within the HOVON-65/GMMG-HD4 trial, we identified 233 patients with 2 copies (67.7%), 95 patients with 3 copies (27.6%) and 16 patients (4.7%) with more than three copies of the chromosomal region 1q21. In addition to del(17p13) and t(4;14), we added +1q21 (>3 copies) to the group of high-risk aberrations, since the outcome of these patients was almost as poor as it was observed for patients with del(17p13). Subsequently, we analyzed whether combining the ISS score with information on the presence of high-risk aberrations could improve the prognostic value with regard to patients' outcome. A combination of the presence or absence of del(17p13), t(4;14), or +1q21 (>3 copies) with the ISS score allowed patients to be stratified into three distinct groups: low-risk [absence of del(17p13)/t(4;14)/+1q21 (>3 copies) and ISS I], high-risk [presence of del(17p13)/t(4;14)/+1q21 (>3 copies) and ISS II/III], and intermediate-risk (all remaining patients). Most of the patients belonged to the low- (33%) and intermediate-risk (49%) groups, whereas 18% were allocated to the high-risk group. The median PFS times for the low-, intermediate-, and high-risk groups were 41.9 months, 31.1 months (HR=1.7; p=0.0018) and 18.7 months (HR=3.6; p These results were confirmed in the independent cohort of patients: From date of first ASCT, the median PFS times for the low-, intermediate-, and high-risk groups were 43.3 months, 23.0 months (HR=1.5; p=0.015) and 13.8 months (HR=2.4; p=0.0003), respectively. The 4yr-OS decreased from 84% in the low-risk group to 71% (HR=2.1; p=0.0043) and 49% (HR=3.84; p CONCLUSION: In our series, the ISS/FISH-based score/algorithm predicted PFS and OS much better than the ISS alone. Our results with molecular cytogenetic techniques may already have implications for the risk-adapted clinical management of patients with MM particularly in younger patients. Disclosures: van de Velde: Ortho Biotech Oncology Research & Development: Employment. Sonneveld:Celgene: Membership on an entity's Board of Directors or advisory committees, Research Funding; Janssen-Cilag: Membership on an entity's Board of Directors or advisory committees, Research Funding; Onyx: Membership on an entity's Board of Directors or advisory committees, Research Funding; Millennium: Membership on an entity's Board of Directors or advisory committees.

Abstract 255 Introduction: Standard treatment of acute myeloid leukemia (AML) comprises one or two cycles of chemotherapy to induce complete remission (CR) followed by postremission treatment in order to prevent relapse of the disease (consolidation therapy). In 2003, we initiated a prospective multicenter randomized trial to investigate the impact of different consolidation strategies on long-term outcome in AML patients ≤ 60 years. Consolidation options comprised upfront allogeneic stem cell transplantation (allo SCT) in aplasia after induction therapy, autologous SCT, and three cycles of standard high-dose-cytarabine-based consolidation. For patients receiving high-dose cytarabine, the main study aim was to evaluate the benefit of adding additional mitoxantrone and amsacrine to cytarabine consolidation. Design: From 2003 to 2009, 1182 patients (median age, 48 years; range 16–60 years) with untreated AML were randomly assigned at diagnosis to different consolidation strategies after classical 7+3 induction. According to the risk-adapted treatment strategy of the trial, cytogenetically or molecular intermediate-risk (IR) and adverse-risk (AR) patients should receive an allo SCT as consolidation treatment if an HLA-identical-sibling donor (IR) or HLA-matched related or unrelated donor (AR) was available. IR and AR patients with no available donor should receive autologous SCT. All favorable risk patients and patients with no available donor were scheduled for high-dose cytarabine based consolidation. Half of the patients were randomized for high dose cytarabine based consolidation. Half of the patients were randomized for high dose cytarabine alone while the other half received high dose cytarabine with the addition of amsacrine and mitoxantrone. Standard chemotherapy consisted of three cycles with high dose cytarabine (2 × 3 g/m2, day 1,3,5) whereas combined consolidation contained two cycles of MAC (cytarabine 2 × 1g/m2, day 1–6, mitoxantrone 10 mg/m2, day 4–6) plus one cycle of MAMAC (cytarabine 2 × 1 g/m2, day 1–5, amsacrine 100 mg/m2, day 1–5). In order to evaluate the effect of the two cytarabine based consolidation strategies, we determined overall survival (OS) and event free survival (EFS) using the method of Kaplan Meyer. Survival distributions were compared using the log rank test. Results: 1182 patients were randomized for further intervention (Arm A+B: n=582, 49.3%; Arm C+D: n=600, 50.7 %). Median follow-up was 41.4 months (95%-CI 39.3–43.6). A total number of 375 patients received allogeneic (n=322) or autologous SCT (n=53) and 807 patients were consolidated with cytarabine. Of these patients, 407 were randomized for cytarabine alone and 400were randomized to receive cytarabine plus mitoxantrone and amsacrine (MAC/MAC/MAMAC). Complete remission rate (CR) after second induction therapy was 59.1% (n=698). Between the four arms, there were no significant differences of the CR rates. Five-year OS of patients receiving high dose cytarabine alone was 47.1% (95%-CI 42.0–52.2%), for patients receiving MAC/MAMAC as consolidation therapy it was 46.8% (95%-CI 42.3–51.3%; p = 0.610). Three-year event free survival (EFS) was also not significant with 30.5% (95%-CI 26.6–34.4%) for patients receiving high dose cytarabine alone and 35.6% (95%-CI 31.7–39.5%; p = 0.059) for patients receiving MAC/MAMAC. Conclusions: According to our data, the addition of mitoxantrone and amsacrine to high dose cytarabine consolidation confers no benefit for treatment outcome in younger AML patients. Disclosures: No relevant conflicts of interest to declare.

Abstract 305 Chromosomal aberrations are important prognostic parameters in multiple myeloma (MM). By using interphase fluorescent in situ hybridization (FISH) on CD138-enriched plasma cells, specific changes in interphase cells can be detected, overcoming the lack of dividing cells required for conventional cytogenetics. We evaluated the association of FISH results and outcome of a subgroup of patients (pts) within the HOVON-65/GMMG-HD4 trial, a prospective, randomized phase III trial for pts with newly diagnosed MM stage II or III according to Salmon & Durie up to 65 years. Pts were randomized to receive three cycles of VAD (arm A; vincristine, adriamycin, dexamethasone) or PAD (arm B; bortezomib, adriamycin, dexamethasone). Hematopoietic stem cells were mobilized using the CAD regimen and G-CSF. All pts received one or two cycles of high dose melphalan (200 mg/m2) with autologous stem cell transplantation followed by maintenance therapy with thalidomide 50 mg daily (arm A) or bortezomib 1.3 mg/m2 once every 2 weeks (arm B), respectively, for a maximum of 2 years. Sites in Germany, the Netherlands and Belgium participated in this trial (n=833 pts). For the German pts (GMMG, n=399) FISH was performed in a single laboratory prior to start of treatment. Cytospins of CD138 purified plasma cells were subjected to FISH with two-color probe sets for the detection of numerical changes for the following chromosome regions: 1q21/8p21, 6q21/15q22, 9q34/22q11, 11q23/13q14, and 17p13/19q13, as well as for the translocations t(4;14)(p16;q32), t(11;14)(q13;q32), and t(14;16) (q32;q23). As of July 2010 data from the first consecutively enrolled 626 patients of the trial have been analyzed, including 284 GMMG pts. For this analysis, FISH results from 258 (91%) GMMG pts were available (n=131 in arm A; n=127 in arm B). Patient characteristics in the GMMG subgroup are comparable to the analyzed trial population of 626 pts. For all pts the median follow-up time from randomization was 40.3 months (mo.). The most pronounced impact on prognosis was seen for t(4;14), del17p13, and gain1q21, each significantly associated with poor prognosis with respect to progression free survival (PFS) and overall survival (OS). The strongest prognostic impact was found for del17p13. FISH analysis detected del17p13 in 9.4% of pts (A: 12.3% vs. B: 6.4%), t(4;14) in 14.8 % (16.3% vs. 13.4%), and gain 1q21 in 33.7% of pts (33.1% vs. 34.4%). When comparing pts in the two arms for PFS, we found a borderline significance for the interaction between t(4;14) and treatment arm (p=0.06), indicating that the effect of t(4;14) depends on the treatment. Pts with t(4;14) in arm A show a very poor prognosis with a median PFS time only half as long compared to patients without translocation (18 vs. 36 mo.; p=0.003). No such negative effect was observed for patients in arm B with t(4;14) (36 vs. 40 mo.; p=0.97). PAD resulted in improved 3yr-OS rates for pts with t(4;14) (A:39% vs B:76%), while 3yr-OS was 79% and 87% in pts without t(4;14). Median PFS for pts with gain 1q21 was 22 mo. (arm A) vs. 30 mo. (arm B) compared to 41 mo. in both arms for patients without gain 1q21. Pts with gain 1q21 showed a significantly better OS when treated with bortezomib (3yr-OS rates: A: 59%, B: 83%, p=0.016). Del17p13 was significantly associated with poor prognosis in both arms for OS (A: p In conclusion our analysis confirms the significant negative prognostic impact of del17p13 and gain 1q21 on PFS and OS for pts with newly diagnosed MM. In our analysis, the negative impact of t(4;14) on PFS could almost completely be overcome by the bortezomib-based treatment. Pts with t(4;14) also showed improved OS when treated with bortezomib, but still had an inferior prognosis compared to pts without this cytogenetic aberration. Similarly, bortezomib significantly prolonged OS in patients with gain 1q21 but did only partially overcome the adverse effect of this aberration. The HOVON65/GMMG-HD4 trial (EudraCT nr 2004-000944-26) was supported by the Dutch Cancer Foundation, the German Federal Ministry of Education and Research and a grant from Janssen Cilag. The GMMG also received grants for this trial by Novartis, AMGEN, Chugai, Roche and the Tumorzentrum Heidelberg/Mannheim. Disclosures: Goldschmidt: Ortho Biotech: Honoraria, Research Funding; Amgen: Research Funding; Novartis: Honoraria, Research Funding; Chugai: Honoraria, Research Funding; Roche: Honoraria, Research Funding; Celgene: Honoraria, Research Funding. Off Label Use: bortezomib in newly diagnosed myeloma. Neben:Ortho Biotech: Honoraria; Celgene: Honoraria. Pfreundschuh:Roche: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Pfizer: Membership on an entity's Board of Directors or advisory committees. Duehrsen:Ortho Biotech: Honoraria. Weisel:Ortho Biotech: Consultancy, Honoraria; Celgene: Consultancy, Honoraria, Research Funding. Scheid:Ortho Biotech: Honoraria. Velde:Johnson & Johnson: Employment. Schmidt-Wolf:Janssen Cilag: financial support (symposium); Celgene: Membership on an entity's Board of Directors or advisory committees. Sonneveld:Johnson & Johnson: Membership on an entity's Board of Directors or advisory committees, Research Funding; Celgene: Membership on an entity's Board of Directors or advisory committees; Millenium: Membership on an entity's Board of Directors or advisory committees.

We report on thermo- and pH-responsive copolymer films consisting of N-isopropylacrylamide (NIPAAm) and different carboxyalkylacrylamides exhibiting switchable swelling states over a wide range of temperatures and pH-values. The copolymers were covalently grafted as brush-like films onto solid substrates. Analysis by quartz crystal microbalance with dissipation monitoring revealed the phase transition behaviour to be sensitively controlled by the balance of polar and non-polar interactions, which could be dissected into the local level of comonomer structure as well as the more global level of copolymer chain composition. Remarkably, the phase transition was determined to be in general lower than in solution, which originated from the higher local polymer concentration of the swollen surface-grafted copolymer layer as explained by the general phase behaviour based on classical theory of polymers in solutions. The different swelling profiles of the environmentally responsive copolymer films are expected to offer valuable options for the design of switchable interfaces as, for instance, applicable for the modulation of model biosystems.

Abstract 3351 Poster Board III-239 Objectives: The majority of patients with chronic lymphocytic leukemia (CLL) who receive allogeneic hematopoietic cell transplantation (HCT) have fludarabine-refractory disease. The most active single agent in this disease stage is alemtuzumab. Alemtuzumab has a long half-life and induces profound T-cell depletion (TCD). Since TCD may mitigate graft-versus leukemia effects we evaluated „pre-conditioning“ with alemtuzumab followed by a washout period in order to minimize in vivo T-cell depletion of the graft in a phase II study (NCT 00337519). Methods: Patients received cytoreductive treatment with 3 × 30 mg alemtuzumab weekly prior to HCT. The scheduled interval between last dose of alemtuzumab and HCT was increased from two weeks to one month during the study. The antibody level at the day of HCT was measured with an ELISA with a lower limit of detection of 31.25 ng/mL (BioAnaLab lim., Oxford, UK). The conditioning regimen contained fludarabine (150 mg/m2) and busulfan (8 mg/kg). Cyclosporine (CSA) and methotrexate (MTX) were applied as GVHD-prophylaxis. Medically fit patients with relapsed CLL were elible. Results: 62 patients with a median age of 57 years were included between April, 2004 and October, 2008. A median of 3 prior regimens had been given. 55% of the patients had fludarabine-resistant disease. Two patients failed to reach HCT due to progressive disease during alemtuzumab therapy. Donors were matched siblings for 26 and matched unrelated donors for 34 patients. The median level of alemtuzumab in peripheral blood after a washout period of two weeks was 62 ng/mL (interquartile range, 45 to 196 ng/mL; minimum below the limit of detection; maximum 490 ng/mL) compared to a median level below the limit of detection after a delay of four weeks (interquartile range, between the limit of detection and 77 ng/mL, maximum 256 ng/mL) (p=0.005). Despite one month time between the last dose of alemtuzumab and HCT 4 out of 30 patients (13%) had alemtuzumab levels greater than 200 ng/mL. No primary or secondary graft failure occurred. A linear relationship between the alemtuzumab level at HCT and the time to complete CD4-T-cell chimerism (TCC) was observed (p=0.003). At day +100 a CD4 positive T-cell-chimerism (TCC) >95% had been achieved by 84% of patients with alemtuzumab levels 200 ng/mL (p=0.006). All patients had a complete neutrophil-chimerism at day +100. After early taper of immunosuppression (N=2) or the application of donor lymphocyte infusions in incremental doses (N=5) mixed TCC has been converted to complete TCC in all patients. The median follow-up is 17 months (1 to 61 months). Day +100 non-relapse mortality was 2%. At two years non-relapse mortality and relapse incidence were 21% and 29%, respectively. Two-year overall survival and progression-free survival were 67% (95% CI, 51% to 83%) and 50% (95% CI, 31% to 69%). Conclusions: In patients who received alemtuzumab prior to HCT, residual drug levels may interfere with T-cell engraftment. Lineage specific T-cell chimerism should therefore be assessed prospectively in this group of patients. Persistent mixed T-cell chimerism can be converted by an early taper of immunosuppression and incremental doses of donor lymphocyte infusions. Disclosures: Schetelig: Bayer Schering: Research Funding. Platzbecker:Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding.

131 Introduction. Autologous stem cell transplantation (ASCT) after cytoreductive induction is considered standard of care for younger patients (pts) with multiple myeloma (MM). The previous standard of induction, the Vincristin-Adriamycin-Dexamethasone (VAD) combination, achieves inferior results compared with induction regimens which combine the proteasome inhibitor Velcade (V = Bortezomib) with Dexamethasone (D)(=VD) and a cytostatic drug such as Doxorubicin (PAD = VD plus Doxorubicin). Velcade-based induction therapy was shown to translate into better myeloma control after high dose melphalan and to lead to prolonged progression-free survival. In order to find a more efficacious and safer drug combination for induction therapy in MM, we tested the combination of Velcade with Cyclophosphamide and Dexamethasone (VCD). Methods. This trial was designed as an open, prospective, multi-center, uncontrolled, combined phase II/III study. As previously reported (Kropff M et al., Ann Hematol 2009), in the first 30 pts the optimal dose of iv Cyclophosphamide in combination with V and D was defined as 900 mg/m2 on d1. Between 03/2006 and 03/2009 we enrolled an additional 370 pts up to 60 years of age with untreated MM to receive three 3-week cycles of induction treatment with V 1.3 mg/m2 iv d1,4,8,11; D 40 mg/d orally d1,2,4,5,8,9,11,12; and C 900mg/m2 iv d1 before scheduled high dose melphalan and ASCT. The primary endpoint of the study is response rate on day 63 after 3 cycles of VCD according to EBMT and IMWG criteria. Results. Final data from 400 pts from 39 German centers will be presented at the meeting. In the currently evaluable 300 pts (mean age 52.3 years; 1.7% stage I, 21.3% stage II, 77.0% stage III) molecular cytogenetic analysis showed a prevalence of 13q- in 38%, of t[4;14] in 13% and of 17p- in 12% of pts (no changes in 35%). All 300 pts (88.3% of whom completed three cycles) were included in the intent-to-treat analysis. Overall response rate (ORR = CR+PR) was 84%, with 10% CR and 74% PR, 5.7% MR, 7.3% NC and 2.3% PD. The negative prognostic impact of 13q- or t[4;14] was abrogated (ORR normal 87.3%, 13q- 83.7%, t[4;14] 90.0%), the unfavorable influence of p53 loss in the 17p- subgroup was still detectable (ORR 69.2%) but this did not reach statistical significance. VGPR rates will be reported at the meeting. Serious adverse events were documented in 78/300 (26.0%) patients. Death rate was remarkably low (1.3%, of which one was not related to the trial medication). 155/300 (52%) of pts experienced grade 3/4 non-serious AEs and of these leucopenia (93/300 pts= 31%), thrombocytopenia (7%), neutropenia (6%), anaemia (5%) were the most frequent events. 80 AEs grade 3 or 4 and 45 SAEs were of infectious origin and occurred in 47/300 pts. 80/130 SAEs (61.5%) were at least possibly related to Velcade. 101/300 pts (34%) developed episodes of peripheral neuropathy. PNP was grade 1 in 62/300 pt (20.7%), grade 2 in 31/300 pt (10.3%) and grade 3 in 7/300 pts (2.3%). Conclusion. This analysis demonstrates that proteasome inhibition by Velcade in combination with Dexamethasone and iv Cyclophosphamide (VCD) is an induction regimen for newly diagnosed MM which is highly effective in a short period of time, has a rather low toxicity profile and is feasible for administration in an outpatient setting. Based on these characteristics, VCD qualifies to become a new standard for MM induction therapy. | Response to VCD | % | |:---------------:| --- | | ORR | 84 | | CR | 10 | | PR | 74 | | MR | 5.7 | | SD | 7.3 | | PD | 2.3 | Table 1: Response to study therapy (intent-to-treat set, according to investigator, n=300) | | Responding patients (≥ PR) | |:-------------------:| -------------------------- | ---- | | n | % | | No FISH abnormality | 69/79 | 87.3 | | 13q- | 72/86 | 83.7 | | t(4;14) | 27/30 | 90 | | 17p- | 18/26 | 69.2 | | Other | 68/77 | 88.3 | Table 2: Response by result of cytogenetic analysis (intent-to-treat set, according to investigator, n=300) Disclosures: Einsele: OrthioBiotech: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding. Off Label Use: Bortezomib is licensed as monotherapy for use in relapsed/refractory MM and in combination with melphalan/Prednisone in the first-line treatment of MM pts ineligible for HD-MEL and ASCT. . Liebisch: OrthoBiotech: Consultancy, Honoraria. Langer: OrthoBiotech: Consultancy. Kropff: OrthoBiotech: Consultancy, Honoraria. Kroger: OrthoBiotech: Honoraria. Ostermann: OrthoBiotech: Honoraria. Mugge: OrthoBiotech: Honoraria. Wolf: OrthoBiotech: Honoraria. Gramatzki: OrthoBiotech: Consultancy, Honoraria. Maschmeyer: OrthoBiotech: Travel Grant. Sezer: OrthoBiotech: Consultancy, Honoraria. Heidemann: OrthoBiotech: Honoraria. Jager: OrthoBiotech: Honoraria. Dechow: Celgene: Research Funding. Simon: OrthoBiotech: Honoraria. Straka: OrthoBiotech: Consultancy, Honoraria, Research Funding. Fingerle-Rowson: orthoBiotech: Employment. Knop: OrthoBiotech: Honoraria.