Description: Summary Meteor Cruise M81/1 was dedicated to the investigation of the distribution of dissolved and particulate trace metals and their isotopic compositions (TEIs) in the full water column of the tropical Atlantic Ocean and their driving factors including main external inputs and internal cycling and ocean circulation. The research program is embedded in the international GEOTRACES program (e.g. Henderson et al., 2007), which this cruise was an official part of and thus corresponds to GEOTRACES cruise GA11. This cruise was completely dedicated to the trace metal clean and contamination-free sampling of waters and particulates for subsequent analyses of the TEIs in the home laboratories of the national and international participants. Besides a standard rosette for the less contaminant prone metals, trace metal clean sampling was realized by using a dedicated and coated trace metal clean rosette equipped with Teflon-coated GO-FLO bottles operated via a polyester coated cable from a mobile winch that was thankfully made available by the U.S. partners of the GEOTRACES program for this cruise. The particulate samples were also collected under trace metal clean conditions using established in-situ pump systems. The cruise track led the cruise southward from the Canary Islands to 11°S and then continued northwestward along the northern margin of South America until it reached Port of Spain, Trinidad & Tobago. The track crossed areas of major external inputs including exchange with the volcanic Canary Islands, the Saharan dust plume, as well as the plume of the Amazon outflow. In terms of internal cycling the equatorial high biological productivity band, as well as increased productivity associated with the Amazon Plume were covered. All major water masses contributing the Atlantic Meridional Overturning Circulation, as well as the distinct narrow equatorial surface and subsurface east-west current bands were sampled. A total of 17 deep stations were sampled for the different dissolved TEIs, which were in most cases accompanied by particulate sampling. In addition, surface waters were continuously sampled under trace metal clean conditions using a towed fish.

Description: Purpose: We previously showed that targeting Delta-like ligand 4 (DLL4) in colon and breast tumors inhibited tumor growth and reduced tumor initiating cell frequency. In this report, we have extended these studies to pancreatic cancer and probed the mechanism of action in tumor and stromal cells involved in antitumor efficacy. Experimental Design: Patient-derived pancreatic xenograft tumor models were used to evaluate the antitumor effect of anti-DLL4. To investigate the mechanism of action, we compared the activity of targeting DLL4 in tumor cells with an anti-human DLL4 antibody (anti-hDLL4) and in the host stroma/vasculature with an anti-mouse DLL4 antibody (anti-mDLL4). The effect of these antibodies on cancer stem cell frequency was examined by in vivo limiting dilution assays. Results: The combination of anti-hDLL4 and anti-mDLL4 was efficacious in a broad spectrum of pancreatic tumor xenografts and showed additive antitumor activity together with gemcitabine. Treatment with either anti-hDLL4 or anti-mDLL4 delayed pancreatic tumor recurrence following termination of gemcitabine treatment, and the two together produced an additive effect. Anti-hDLL4 had a pronounced effect in reducing the tumorigenicity of pancreatic cancer cells based on serial transplantation and tumorsphere assays. In contrast, disruption of tumor angiogenesis with anti-mDLL4 alone or with anti-VEGF had minimal effects on tumorigenicity. Gene expression analyses indicated that anti-DLL4 treatment regulated genes that participate in Notch signaling, pancreatic differentiation, and epithelial-to-mesenchymal transition. Conclusions: Our findings suggest a novel therapeutic approach for pancreatic cancer treatment through antagonism of DLL4/Notch signaling. Clin Cancer Res; 18(19); 5374–86. ©2012 AACR .

Description: The GANDALF 6U-VME64x/VXS module has been developed for the digitization and real time analysis of detector signals. To perform different applications such as analog-to-digital or time-to-digital conversions, coincidence matrix formation, fast pattern recognition and trigger generation, this module comes with exchangeable analog and digital mezzanine cards. Based on this platform, we present a 128-channel TDC which is implemented in a single Xilinx Virtex-5 FPGA using a shifted clock sampling method. In contrast to common TDC concepts, the input signal is sampled by 16 equidistant phase-shifted clocks. A particular challenge of the design is the minimum skew routing of the input signals to the sampling flip-flops. We present measurement results for the differential nonlinearity and the time resolution of the TDC readout system.

Description: Aims The dispersal of pollen and seeds is spatially restricted and may vary among plant populations because of varying biotic interactions, population histories or abiotic conditions. Because gene dispersal is spatially restricted, it will eventually result in the development of spatial genetic structure (SGS), which in turn can allow insights into gene dispersal processes. Here, we assessed the effect of habitat characteristics like population density and community structure on small-scale SGS and estimate historical gene dispersal at different spatial scales. Methods In a set of 12 populations of the subtropical understory shrub Ardisia crenata , we assessed genetic variation at 7 microsatellite loci within and among populations. We investigated small-scale genetic structure with spatial genetic autocorrelation statistics and heterogeneity tests and estimated gene dispersal distances based on population differentiation and on within-population SGS. SGS was related to habitat characteristics by multiple regression. Important Findings The populations showed high genetic diversity ( H e = 0.64) within populations and rather strong genetic differentiation ( $${{F}^{\prime }}_{\hbox{ ST }}$$ = 0.208) among populations, following an isolation-by-distance pattern, which suggests that populations are in gene flow–drift equilibrium. Significant SGS was present within populations (mean Sp = 0.027). Population density and species diversity had a joint effect on SGS with low population density and high species diversity leading to stronger small-scale SGS. Estimates of historical gene dispersal from between-population differentiation and from within-population SGS resulted in similar values between 4.8 and 22.9 m. The results indicate that local-ranged pollen dispersal and inefficient long-distance seed dispersal, both affected by population density and species diversity, contributed to the genetic population structure of the species. We suggest that SGS in shrubs is more similar to that of herbs than to trees and that in communities with high species diversity gene flow is more restricted than at low species diversity. This may represent a process that retards the development of a positive species diversity–genetic diversity relationship.

Description: Dengue virus has emerged as a global health threat to over one-third of humankind. As a positive-strand RNA virus, dengue virus relies on the host cell metabolism for its translation, replication, and egress. Therefore, a better understanding of the host cell metabolic pathways required for dengue virus infection offers the opportunity to develop new approaches for therapeutic intervention. In a recently described screen of known drugs and bioactive molecules, we observed that methotrexate and floxuridine inhibited dengue virus infections at low micromolar concentrations. Here, we demonstrate that all serotypes of dengue virus, as well as West Nile virus, are highly sensitive to both methotrexate and floxuridine, whereas other RNA viruses (Sindbis virus and vesicular stomatitis virus) are not. Interestingly, flavivirus replication was restored by folinic acid, a thymidine precursor, in the presence of methotrexate and by thymidine in the presence of floxuridine, suggesting an unexpected role for thymidine in flavivirus replication. Since thymidine is not incorporated into RNA genomes, it is likely that increased thymidine production is indirectly involved in flavivirus replication. A possible mechanism is suggested by the finding that p53 inhibition restored dengue virus replication in the presence of floxuridine, consistent with thymidine-less stress triggering p53-mediated antiflavivirus effects in infected cells. Our data reveal thymidine synthesis pathways as new and unexpected therapeutic targets for antiflaviviral drug development.

Description: Quantitative assessment of growth of filamentous microorganisms, such as streptomycetes, is generally restricted to determination of dry weight. Here, we describe a straightforward methylene blue-based sorption assay to monitor microbial growth quantitatively, simply, and rapidly. The assay is equally applicable to unicellular and filamentous bacterial and eukaryotic microorganisms.

Description: The t(12;21) chromosomal translocation, targeting the gene encoding the RUNX1 transcription factor, is observed in 25% of pediatric acute lymphoblastic leukemia (ALL) and is an initiating event in the disease. To elucidate the mechanism by which RUNX1 disruption initiates leukemogenesis, we investigated its normal role in murine B-cell development. This study revealed 2 critical functions of Runx1: (1) to promote survival and development of progenitors specified to the B-cell lineage, a function that can be substituted by ectopic Bcl2 expression, and (2) to enable the developmental transition through the pre-B stage triggered by the pre-B-cell antigen receptor (pre-BCR). Gene expression analysis and genomewide Runx1 occupancy studies support the hypothesis that Runx1 reinforces the transcription factor network governing early B-cell survival and development and specifically regulates genes encoding members of the Lyn kinase subfamily (key integrators of interleukin-7 and pre-BCR signaling) and the stage-specific transcription factors SpiB and Aiolos (critical downstream effectors of pre-BCR signaling). Interrogation of expression databases of 257 ALL samples demonstrated the specific down-regulation of the SPIB and IKZF3 genes (the latter encoding AIOLOS) in t(12;21) ALL, providing novel insight into the mechanism by which the translocation blocks B-cell development and promotes leukemia.

Description: A group of 400–500 m long, bedding-parallel calcite veins are exposed in the central La Popa Basin of northeastern Mexico. These veins provide a unique opportunity to determine the kilometer-scale fluid–rock system associated with bedding-parallel vein formation, and to test for sampling bias in studies that often use one or two samples to constrain the characteristics of regional-scale paleohydrogeological systems. We use fluid inclusion microthermometry in conjunction with measurements of δ 13 C, δ 18 O, and 87 Sr/ 86 Sr ratios to constrain the vein-forming fluid temperatures, compositions and sources, and compare these values along and between the veins to establish the homogeneity of the vein-forming fluids and fluid–rock system. The δ 13 C values of the veins are close to those of the host rock, and average – 3.96‰ (PDB). The δ 18 O values of the veins are typically 1‰ lower than those of the host rocks, and average – 9.54‰ (PDB). Fluid inclusion homogenization temperatures average 137°C and inclusion salinities are all 〈6 wt% NaCl equivalent. The 87 Sr/ 86 Sr ratios of the veins average 0.70731 and are substantially lower than the values expected for the host rock. Calculated fluid δ 18 O values range from 4 to 10‰ (SMOW). The isotopic and microthermometric data indicate the veins most likely formed at depths of 3–4 km when meteoric water mixed with upward migrating, warm basinal brines. Vein microstructures and field characteristics indicate they formed from multiple slip events that most likely were associated with transport of individual fluid pulses that migrated along bedding planes. The large-scale homogeneity of vein geochemistry is remarkable and demonstrates that only one or two samples would be sufficient to accurately characterize the kilometer-scale paleohydrogeological system for these veins. We conducted petrographic, isotopic, and fluid inclusion analyses on a set of bed-parallel veins that can be traced for over 400 m. The results indicate that the vein-forming fluids were homogeneous and derived from downward-percolating meteoric water and upward-flowing, low salinity, basinal brine. The paleohydrological system that formed the veins was uniform at scales of hundreds of meters to kilometers.

Description: In the autumn of 2011, Schmallenberg virus (SBV), a novel orthobunyavirus of the Simbu serogroup, was identified by metagenomic analysis in Germany. SBV has since been detected in ruminants all over Europe, and investigations on phylogenetic relationships, clinical signs and epidemiology have been conducted. However, until now, only comparative sequence analysis of SBV genome segments with other species of the Simbu serogroup have been performed, and detailed data on the S and M segments, relevant for virus–host-cell interaction, have been missing. In this study, we investigated the S- and M-segment sequences obtained from 24 SBV-positive field samples from sheep, cattle and a goat collected from all over Germany. The results obtained indicated that the overall genome variability of SBV is neither regionally nor host species dependent. Nevertheless, we characterized for the first time a region of high sequence variability (a mutation ‘hot spot’) within the glycoprotein Gc encoded by the M segment.