Terpene Metabolic Engineering Via Nuclear or Chloroplast Genomes Profoundly and Globally Impacts Off‐Target Pathways Through Metabolite Signalling

The impact of metabolic engineering on nontarget pathways and outcomes of metabolic engineering from different genomes are poorly understood questions. Therefore, squalene biosynthesis genes FARNESYL DIPHOSPHATE SYNTHASE (FPS) and SQUALENE SYNTHASE (SQS) were engineered via the Nicotiana tabacum chloroplast (C), nuclear (N) or both (CN) genomes to promote squalene biosynthesis. SQS levels were ~4300‐fold higher in C and CN lines than in N, but all accumulated ~150‐fold higher squalene due to substrate or storage limitations. Abnormal leaf and flower phenotypes, including lower pollen production and reduced fertility, were observed regardless of the compartment or level of transgene expression. Substantial changes in metabolomes of all lines were observed: levels of 65–120 unrelated metabolites, including the toxic alkaloid nicotine, changed by as much as 32‐fold. Profound effects of transgenesis on nontarget gene expression included changes in the abundance of 19 076 transcripts by up to 2000‐fold in CN; 7784 transcripts by up to 1400‐fold in N; and 5224 transcripts by as much as 2200‐fold in C. Transporter‐related transcripts were induced, and cell cycle‐associated transcripts were disproportionally repressed in all three lines. Transcriptome changes were validated by qRT‐PCR. The mechanism underlying these large changes likely involves metabolite‐mediated anterograde and/or retrograde signalling irrespective of the level of transgene expression or end product, due to imbalance of metabolic pools, offering new insight into both anticipated and unanticipated consequences of metabolic engineering

The Magnetohydrodynamics of Shock-Cloud Interaction in Three Dimensions

The magnetohydrodynamic evolution of a dense spherical cloud as it interacts with a strong planar shock is studied, as a model for shock interactions with density inhomogeneities in the interstellar medium. The cloud is assumed to be small enough that radiative cooling, thermal conduction, and self-gravity can be ignored. A variety of initial orientations (including parallel, perpendicular, and oblique to the incident shock normal) and strengths for the magnetic field are investigated. During the early stages of the interaction (less than twice the time taken for the transmitted shock to cross the interior of the cloud) the structure and dynamics of the shocked cloud is fairly insensitive to the magnetic field strength and orientation. However, at late times strong fields substantially alter the dynamics of the cloud, suppressing fragmentation and mixing by stabilizing the interface at the cloud surface. Even weak magnetic fields can drastically alter the evolution of the cloud compared to the hydrodynamic case. Weak fields of different geometries result in different distributions and amplifications of the magnetic energy density, which may affect the thermal and non-thermal x-ray emission expected from shocked clouds associated with, for example, supernovae remnants.Comment: Accepted for publication in Astrophysical Journal; a higher resolution file can be found at http://www.astro.princeton.edu/~msshin/science/shock_cloud.pdf.g

Terpene Metabolic Engineering \u3cem\u3evia\u3c/em\u3e Nuclear or Chloroplast Genomes Profoundly and Globally Impacts Off-Target Pathways Through Metabolite Signalling

The impact of metabolic engineering on nontarget pathways and outcomes of metabolic engineering from different genomes are poorly understood questions. Therefore, squalene biosynthesis genes FARNESYL DIPHOSPHATE SYNTHASE (FPS) and SQUALENE SYNTHASE (SQS) were engineered via the Nicotiana tabacum chloroplast (C), nuclear (N) or both (CN) genome to promote squalene biosynthesis. SQS levels were ~4300-fold higher in C and CN lines than in N, but all accumulated ~150-fold higher squalene due to substrate or storage limitations. Abnormal leaf and flower phenotypes, including lower pollen production and reduced fertility, were observed regardless of the compartment or level of transgene expression. Substantial changes in metabolomes of all lines were observed: levels of 65-120 unrelated metabolites, including the toxic alkaloid nicotine, changed by as much as 32-fold. Profound effects of transgenesis on nontarget gene expression included changes in the abundance of 19 076 transcripts by up to 2000-fold in CN; 7784 transcripts by up to 1400-fold in N; and 5224 transcripts by as much as 2200-fold in C. Transporter-related transcripts were induced, and cell cycle-associated transcripts were disproportionately repressed in all three lines. Transcriptome changes were validated by qRT-PCR. The mechanism underlying these large changes likely involves matabolite-mediated anterograde and/or retrograde signalling irrespective of the level of transgene expression or end product, due to imbalance of metabolic pools, offering new insight into both anticipated and unanticipated consequences of metabolic engineering

Maximum likelihood analysis of systematic errors in interferometric observations of the cosmic microwave background

We investigate the impact of instrumental systematic errors in interferometric measurements of the cosmic microwave background (CMB) temperature and polarization power spectra. We simulate interferometric CMB observations to generate mock visibilities and estimate power spectra using the statistically optimal maximum likelihood technique. We define a quadratic error measure to determine allowable levels of systematic error that do not induce power spectrum errors beyond a given tolerance. As an example, in this study we focus on differential pointing errors. The effects of other systematics can be simulated by this pipeline in a straightforward manner. We find that, in order to accurately recover the underlying B-modes for r=0.01 at 28<l<384, Gaussian-distributed pointing errors must be controlled to 0.7^\circ rms for an interferometer with an antenna configuration similar to QUBIC, in agreement with analytical estimates. Only the statistical uncertainty for 28<l<88 would be changed at ~10% level. With the same instrumental configuration, we find the pointing errors would slightly bias the 2-\sigma upper limit of the tensor-to-scalar ratio r by ~10%. We also show that the impact of pointing errors on the TB and EB measurements is negligibly small.Comment: 10 pages, 4 figures, accepted for publication in ApJS. Includes improvements in clarity of presentation and Fig.4 added, in response to refere

Genetic ablation or chemical inhibition of phosphatidylcholine transfer protein attenuates diet?induced hepatic glucose production†‡

 Phosphatidylcholine transfer protein (PC?TP, synonym StARD2) is a highly specific intracellular lipid binding protein that is enriched in liver. Coding region polymorphisms in both humans and mice appear to confer protection against measures of insulin resistance. The current study was designed to test the hypotheses that Pctp?/? mice are protected against diet?induced increases in hepatic glucose production and that small molecule inhibition of PC?TP recapitulates this phenotype. Pctp?/? and wildtype mice were subjected to high?fat feeding and rates of hepatic glucose production and glucose clearance were quantified by hyperinsulinemic euglycemic clamp studies and pyruvate tolerance tests. These studies revealed that high?fat diet?induced increases in hepatic glucose production were markedly attenuated in Pctp?/? mice. Small molecule inhibitors of PC?TP were synthesized and their potencies, as well as mechanism of inhibition, were characterized in vitro. An optimized inhibitor was administered to high?fat?fed mice and used to explore effects on insulin signaling in cell culture systems. Small molecule inhibitors bound PC?TP, displaced phosphatidylcholines from the lipid binding site, and increased the thermal stability of the protein. Administration of the optimized inhibitor to wildtype mice attenuated hepatic glucose production associated with high?fat feeding, but had no activity in Pctp?/? mice. Indicative of a mechanism for reducing glucose intolerance that is distinct from commonly utilized insulin?sensitizing agents, the inhibitor promoted insulin?independent phosphorylation of key insulin signaling molecules. Conclusion: These findings suggest PC?TP inhibition as a novel therapeutic strategy in the management of hepatic insulin resistance

Breast cancer derived GM-CSF regulates arginase 1 in myeloid cells to promote an immunosuppressive microenvironment

Tumor-infiltrating myeloid cells contribute to the development of the immunosuppressive tumor microenvironment. Myeloid cell expression of arginase 1 (Arg-1) promotes a protumor phenotype by inhibiting T cell function and depleting extracellular L-arginine, but the mechanism underlying this expression, especially in breast cancer, is poorly understood. In breast cancer clinical samples and in our mouse models, we identified tumor derived GM-CSF as the primary regulator of myeloid cell Arg-1 expression and local immune suppression through a gene knockout screen of breast tumor cell-produced factors. The induction of myeloid cell Arg-1 required GM-CSF and a low pH environment. GM-CSF signaling through STAT3, p38 MAPK, and acid signaling through cAMP were required to activate myeloid cell Arg-1 expression in a STAT6 independent manner. Importantly, breast tumor cell-derived GM-CSF promoted tumor progression by inhibiting host anti-tumor immunity, driving a significant accumulation of Arg-1 expressing myeloid cells compared to lung and melanoma tumors with minimal GM-CSF expression. Blockade of tumoral GM-CSF enhanced the efficacy of tumor-specific adoptive T-cell therapy and immune checkpoint blockade. Taken together, breast tumor cell-derived GM-CSF contributes to the development of the immunosuppressive breast cancer microenvironment by regulating myeloid cell Arg-1 expression and can be targeted to enhance breast cancer immunotherapy

Oligocene deformation of the Chuandian terrane in the SE margin of the Tibetan Plateau related to the extrusion of Indochina

Mechanisms driving the tectonic evolution of the southeast (SE) margin of Tibet include the Paleogene extrusion of the coherent Indochina lithospheric block, and the continuous deformation caused by lower crustal flow since the middle Miocene. The timing and style of regional deformations are key to determining the role of each mechanism. Fault-bounded and -controlled Cenozoic basins within the SE margin of Tibet record regional deformation, surface uplift and variations in paleoclimate, but often are poorly dated. New magnetostratigraphy and 40Ar/39Ar dating of volcanic ashes constrain precisely the timing of sedimentation within the Lühe Basin to between ~35 and 26.5 Ma. The basin is located in the Chuandian terrane along the Chuxiong fault, which lies ~70 km north of, and parallel to, the Ailao Shan-Red River fault. The asymmetric syncline of the Lühe Basin suggests syn-contractional sedimentation and the basal age of the basin represents the initiation of the Chuxiong fault and crustal shortening at ~35 Ma. This is coincident with the onset of the Ailao Shan-Red River fault, and supports a kinematic link between them. Our study suggests that, like the Ailao Shan-Red River fault, the Chuxiong fault is a Paleogene transpressional structure that developed during the extrusion and clockwise rotation of Indochina around the Eastern Himalayan Syntaxis, which caused the late Paleogene deformation and surface uplift of the Chuandian terrane and Indochina. Our revised chronostratigraphy of the Lühe Basin provides further evidence that many of the “Neogene” sedimentary basins in the SE margin of Tibet may be much older than previously thought

Pacific Northwest National Laboratory Site Environmental Report for Calendar Year 2011

The PNNL Site Environmental Report for Calendar Year 2011 was prepared pursuant to the requirements of Department of Energy (DOE) Order 231.1B, "Environment, Safety and Health Reporting" to provide a synopsis of calendar year 2011 information related to environmental management performance and compliance efforts. It summarizes site compliance with federal, state, and local environmental laws, regulations, policies, directives, permits, and orders and environmental management performance

Spiral spin-liquid and the emergence of a vortex-like state in MnSc2_2S4_4

Spirals and helices are common motifs of long-range order in magnetic solids, and they may also be organized into more complex emergent structures such as magnetic skyrmions and vortices. A new type of spiral state, the spiral spin-liquid, in which spins fluctuate collectively as spirals, has recently been predicted to exist. Here, using neutron scattering techniques, we experimentally prove the existence of a spiral spin-liquid in MnSc$_2$S$_4$ by directly observing the 'spiral surface' - a continuous surface of spiral propagation vectors in reciprocal space. We elucidate the multi-step ordering behavior of the spiral spin-liquid, and discover a vortex-like triple-q phase on application of a magnetic field. Our results prove the effectiveness of the $J_1$-$J_2$ Hamiltonian on the diamond lattice as a model for the spiral spin-liquid state in MnSc$_2$S$_4$, and also demonstrate a new way to realize a magnetic vortex lattice.Comment: 10 pages, 11 figure