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At least 343 records · Page 19

A genome-wide analysis of the flax (Linum usitatissimum L.) dirigent protein family: from gene identification and evolution to differential regulation

Identification of DIR encoding genes in flax genome. Analysis of phylogeny, gene/protein structures and evolution. Identification of new conserved motifs linked to biochemical functions. Investigation of spatio-temporal gene expression and response to stress. Dirigent proteins (DIRs) were discovered during 8-8' lignan biosynthesis studies, through identification of stereoselective coupling to afford either (+)- or (-)-pinoresinols from E-coniferyl alcohol. DIRs are also involved or potentially involved in terpenoid, allyl/propenyl phenol lignan, pterocarpan and lignin biosynthesis. DIRs have very large multigene families in different vascular plants including flax, with most still of unknown function. DIR studies typically focus on a small subset of genes and identification of biochemical/physiological functions. Herein, a genome-wide analysis and characterization of the predicted flax DIR 44-membered multigene family was performed, this species being a rich natural grain source of 8-8' linked secoisolariciresinol-derived lignan oligomers. All predicted DIR sequences, including their promoters, were analyzed together with their public gene expression datasets. Expression patterns of selected DIRs were examined using qPCR, as well as through clustering analysis of DIR gene expression. These analyses further implicated roles for specific DIRs in (-)-pinoresinol formation in seed-coats, as well as (+)-pinoresinol in vegetative organs and/or specific responses to stress. Phylogeny and gene expression analysis segregated flax DIRs into six distinct clusters with new cluster-specific motifs identified. We propose that these findings can serve as a foundation to further systematically determine functions of DIRs, i.e. other than those already known in lignan biosynthesis in flax and other species. Given the differential expression profiles and inducibility of the flax DIR family, we provisionally propose that some DIR genes of unknown function could be involved in different aspects of secondary cell wall biosynthesis and plant defense.

dirigent protein↗

Structure Guided Functional Studies of a Lipoprotein Involved in Salmonella Copper Homeostasis

Cells from aerobic organisms require copper ions to generate biochemical energy and for certain enzyme‐catalyzed redox reactions. However, recent studies suggest that when bacteria are engulfed by phagosomes within macrophages, Cu 2+ is used in high concentrations, among other molecules, to kill the bacterium. To infect a host, bacteria such as Salmonella enterica must survive this toxic environment containing high Cu 2+ levels. A recently discovered protein of unknown function, DcrB, plays a role in copper homeostasis within Salmonella enterica , a major cause of food‐borne illness. DcrB is a small lipoprotein anchored to the periplasmic face of the cytoplasmic membrane. Currently, Salmonella phenotypic assays have shown DcrB is necessary for growth in high levels of Cu 2+ . The hypothesis of this project is that DcrB binds copper and possibly other divalent cations. To test this hypothesis, we used thermal stability assays paired with structure‐based functional studies. We examined the effect of divalent cations on the stability of purified DcrB protein using a fluorescence‐based thermal shift assay and circular dichroism. Furthermore, we are using our recent crystallographic structure of DcrB to identify possible metal ion binding sites and to test whether each hypothesized metal ion binding site is required for the ability of DcrB to confer resistance to Cu 2+ . For each site, we used site‐directed mutagenesis to generate an allele of dcrB that encodes a DcrB variant lacking the functional side‐chains at the site. Then, we determined whether a multicopy plasmid that contains the mutant allele of dcrB can complement the copper sensitivity of a dcrB knockout mutant of Salmonella enterica . Our results have established the specificity of DcrB for binding divalent cations and have led to the identification of potential metal binding sites, which we anticipate will shed light on the role of DcrB in metal homeostasis in pathogenic bacteria. Support or Funding Information This research was supported by a UW‐La Crosse Faculty Research Grant (JFM), a UW‐La Crosse Undergraduate Research Grant (RWS), and the Dean's Distinguished Fellowship (RWS). This research used resources of the Advanced Photon Source, a U.S. Department of Energy Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract No. DEAC0206CH11357. Use of the LS‐CAT Sector 21 was supported by the Michigan Economic Development Corporation and the Michigan Technology Tri‐Corridor (Grant 085P1000817). This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .

Soens, Ross↗

ECC Ozonesonde Reliability, Observations, and Comparisons with Satellite Ozone Instruments

Electrochemical Concentration Cell (ECC) ozone instruments depend on the quality of care exercised in their pre-flight preparation. The ozone-measuring project conducted at Goddard Space Flight Center's Wallops Flight Facility uses a number of mechanisms designed to inspect the ECC for anomalies that may interfere with the reception of valid ozone profiles. Complete electronic testing of the instrument, individually and when coupled to its radiosonde has led to exceptional monitoring of ozone for detecting long-term atmospheric changes. A number of factors are considered when preparing an ECC instrument for flight. These basically are specific calibrations of pump efficiency, volumetric flow rate, temperature of the air entering the pump, and background current. The concentration of the potassium iodide solution is also important. Wallops is the only site using a UV photometer (Dasibi) to compare ECC ozone output at various concentrations of ozone that allows adjustment to be made to offsets that may appear in the balloon-borne instrument prior to release. All of the above procedures allow identification of potential problems before release of the ECC instrument. Procedures followed at Wallops also are employed in Brazil, and Ascension Island where NASA has cooperative agreements in place to obtain ozonesondes data. All ECC instruments are prepared 3-4 weeks prior to the day of observation. We will briefly describe the instrumental tests employed. These tests have included simultaneous dual observations to compare the effect of different solution concentrations, comparison of sensors of different manufacturers, and comparisons with surface- and space-based instrumentation such as the Dobson Spectrophotometer and satellites. Vertical profiles of ozone from Arctic, mid-latitudes, and Antarctica will be discussed. Although not unusual, the data reveals ozone structure that correlate well with typical atmospheric temperatures and possibly relative humidity. Finally, vertical ozone distribution, compared with remotely measured ozone from lidar and satellite, will be discussed. Specific comparisons between ECC and HALOE measurements, integrated ECC total ozone overburden with the EP-TOMS and the Dobson, as well as comparisons with lidar are discussed. Results show agreement and some disagreement between the in situ measurements of the ECC and the remote instruments. We postulate reasons for the differences, or biases, which in spite of the excellent ECC quality control during pre-flight preparation and data analysis processes, may be due to uncertainties in both measuring systems.

Schmidlin, F. J.↗

Structure-guided utilization of lignocellulose for catalysis, energy, and biomaterials

As a complex composite of cellulose, hemicellulose, and lignin, plant lignocellulose has long served as a major resource for biomass conversion, materials engineering, and bio-based product development. High-resolution structural insights enabled by solid-state nuclear magnetic resonance (ssNMR) now allow the mapping of polymer interfaces, identification of functional group accessibility, and tracking of molecular organization during processing, all of which are critical factors for optimizing catalytic strategies. These insights could drive transformative progress in lignocellulose-based applications, including selective depolymerization, improved pretreatment design, and efficient upcycling of lignin into resins, plastics, and biomedical materials. In industry-relevant contexts, such as biofuel generation and renewable material manufacturing, understanding the hydration dynamics, cross-linking patterns, and structural heterogeneity is also essential. The ability to visualize these features in native biomass presents a unique opportunity to develop new strategies for sustainability and performance. As the structural toolbox continues to expand, it is becoming a central enabler for innovations in renewable energy, green chemistry, and advanced bioproducts.

bioproduct↗

Template-Induced Structuring and Tunable Polymorphism of Three-Dimensionally Ordered Mesoporous (3DOm) Metal Oxides

Convectively assembled colloidal crystal templates, comprised of size-tunable (ca. 15-50 nm) silica (SiO 2 ) nanoparticles, enable versatile sacrificial templating of three-dimensionally ordered mesoporous (3DOm) metal oxides (MO x ) at both mesoscopic and microscopic size-scales. Specifically, we show for titania (TiO 2 ) and zirconia (ZrO 2 ) how this approach not only enables the engineering of the mesopore size, pore volume, and surface area but can also be leveraged to tune crystallite polymorphism of the resulting 3DOm metal oxides. Template-mediated volumetric (i.e., interstitial) effects and interfacial factors are shown to preserve the metastable crystalline polymorphs of each corresponding 3DOm oxide (i.e., anatase TiO 2 (A-TiO 2 ) and tetragonal ZrO 2 (t-ZrO 2 )) during high-temperature calcination. Mechanistic investigations suggest that this polymorph stabilization is derived from the combined effects of the templatereplica (MO x /SiO 2 ) interface and simultaneous interstitial confinement that limits the degree of coarsening during high temperature calcination of the template-replica composite. The result is the identification of a facile yet versatile templating strategy for realizing 3DOm oxides with (i) surface areas that are more than an order of magnitude larger than those untemplated control samples, (ii) pore diameters and volumes that can be tuned across a continuum of size-scales, and (iii) selectable polymorphism.

36 MATERIALS SCIENCE↗

Potential Evaporite Biomarkers from the Dead Sea

The Dead Sea is located on the northern branch of the African-Levant Rift systems. The rift system, according to one model, was formed by a series of strike slip faults, initially forming approximately two million years ago. The Dead Sea is an evaporite basin that receives freshwater from springs and from the Jordan River. The Dead Sea is different from other evaporite basins, such as the Great Salt Lake, in that it possesses high concentrations of magnesium and has an average pH of 6.1. The dominant cation in the Great Salt Lake is sodium, and the pH is 7.7. Calcium concentrations are also higher in the Dead Sea than in the Great Salt Lake. Both basins are similar in that the dominant anion is chlorine and the salinity levels are approximately 20 %. Other common cations that have been identified from the waters of the Dead Sea and the Great Salt Lake include sodium and potassium. A variety of Archea, Bacteria, and a single genus of a green algal, Dunaliella, has been described from the Dead Sea. Earlier studies concentrated on microbial identification and analysis of their unique physiology that allows them to survive in this type of extreme environment. Potential microbial fossilization processes, microbial fossils, and the metallic ions associated with fossilization have not been studied thoroughly. The present study is restricted to identifying probable microbial morphologies and associated metallic ions. XRD (X Ray Diffraction) analysis indicates the presence of halite, quartz, and orthoclase feldspar. In addition to these minerals, other workers have reported potassium chloride, magnesium bromide, magnesium chloride, calcium chloride, and calcium sulfate. Halite, calcium sulfate, and orthoclase were examined in this report for the presence of microbes, microbially induced deposits or microbial alteration. Neither the gypsum nor the orthoclase surfaces possesses any obvious indications of microbial life or fossilization. The sand-sized orthoclase particles are weathered with 122 extensive fan-shaped mineral deposits. The gypsum deposits are associated with halite minerals and also exhibit extensive weathering. Halite minerals represent the only substrates that have probable rod-shaped microbial structures with long, filamentous, apical extensions. EDS (energy dispersive x-ray) analysis of the putative microbes indicates elevated calcium levels that are enriched with magnesium. The rod-shaped structures exhibit possible fossilization stages. Rhombohedralshaped minerals of magnesium-enriched calcium carbonate are deposited on the microbial surfaces, and eventually coat the entire microbial surface. The sodium chloride continues to crystallize on nearby halite surface and even crystallizes on the fossilized microbial remains. The putative fossils are found exclusively on halite surfaces, and all contained elevated levels of calcium magnesium cations. Both of these metallic cations are associated with microbial activity and fossilization. Their morphological diversity is low in comparison with the reported living Dead Sea microbial population. If we examine the fossil record for multicellular organisms, fossilization rates are lower for soft-bodied organisms than for those possessing hard parts, i.e. shells, bones. For example, smaller, single celled organisms would have a smaller chance of fossilization; their fossilized shapes could be mistaken for abiotic products. Another consideration is that dead organisms in the water column are probably utilized as a food source by other microbes before fossilization processes are completed. This may be an important consideration as we attempt to model and interpret ancient microbial environments either on Earth or on Mars.

Morris, Penny A.↗

Contribution of Spaceflight Environmental Factors to Vision Risks

The risk of visual impairment and elevated intracranial pressure as a result of low-earth orbit microgravity exposure has directed our attention and research efforts to the eye. While the alterations observed in astronauts returning from long duration missions include vision and neuroanatomical changes observed by non-invasive methods, other effects and subsequent tissue responses at the molecular and cellular level can only be studied by accessing the tissue itself. As a result of this need, several studies are currently taking place within the Human and Health Countermeasures Element (HHC) that use animal models for eye research. The rodent eye has many similarities to the human eye, and both rats and mice have historically been used as models of human eye disease, aiding in the identification of the disease genes, elucidation of mechanisms of disease, as well as in the assessment of therapeutic treatments. These studies attempt to answer two central questions in the etiology of possible vision alterations in the environment of space exploration missions. The first is: what effects and response mechanisms take place in the different eye structures at the cellular and molecular level? The second question is directed to elucidate the contribution of the various environmental stressors (radiation, nutrition, fluid shift) to these effects. Collaborative approaches with internal and external investigators have allowed performing these studies in a most cost-effective fashion, providing preliminary data and laying the bases for testing further hypotheses in future and specifically designed animal experiments. From a study centered on the radioadaptive response in mice, we have learned that the retina responds to low and high dose gamma radiation by elevating antioxidant-related genes at early time points (4hrs) and that this response returns to control levels after 1 day post-irradiation. We are expanding this research with another collaborative study that investigates the combined effects of radiation exposure and iron overload on sensitivity to radiation injury in rat eyes. All main eye structures will be analyzed in this study: retina, lens and cornea. A study in collaboration with the Space Human Factors and Habitability Element (SHFH) investigates the effects of lunar dust exposure on the rat cornea. It is anticipated that common underlying oxidative stress mechanisms of damage may be observed as a result of these three stressors: radiation, nutritional iron and lunar dust. The contribution of fluid shift is addressed by a study using rats subjected to hindlimb suspension. The hypothesis to be tested in this study is that the mechanical stress imparted by the pressure differential across the optic disc and lamina cribosa will impact oxygenation (therefore causing oxidative stress and hypoxia) and cell survival. This study also includes the assessment of two nutritional antioxidant countermeasures: epigallocatechin gallate (green tea) and resveratrol. Finally, as a result of two successful tissue sharing efforts, we are proceeding with the analysis of eye samples of mice aboard two shuttle missions: STS-133 and STS-135. Results from the STS-133 study are presented in an independent abstract. Briefly, the results show that spaceflight represents a source of environmental stress that directly translates into oxidative and cellular stress in the retina. Similar analysis is also planned for the cornea. These samples add large value to our current vision research as they provide data on the direct effects of low-earth orbit spaceflight on eye structures and physiology.

Zanello, Susana B.↗

Estimation of constituent properties of concrete materials with an artificial neural network based method

Multi-scale models are developed for heterogeneous concrete materials to estimate their macroscopic mechanical properties in terms of micro-structural data. One crucial challenge of those models is the identification of local properties of constituent phases. In this paper, we present an efficient method based on Artificial Neural Networks (ANN). Typical concrete materials are taken as example. A macroscopic analytical strength criterion is established from three steps of nonlinear homogenization procedure. The macroscopic strength of materials is determined as a function of the frictional coefficient and cohesion of solid cement particles at nanometer scale, intra-particle pores, inter-particle pores and aggregates (inclusions). The objective is to identify the nanoscopic frictional coefficient and cohesion of cement particle from measured macroscopic values of uniaxial compression and tensile strengths. For this purpose, a numerical method based on the ANN is developed. With the analytical macroscopic strength criterion, sensitivity studies are first realized to identify the most important micro-structural parameters influencing the macroscopic strength of concrete. A simplified analytical macroscopic strength criterion is then proposed. A large dataset is further constructed through the inversion of the analytical strength criterion by using the aggregates volume fraction, porosity, macroscopic uniaxial tensile and compressive strengths as input variables and the frictional coefficient and cohesion of cement particles as output unknowns. An ANN model containing four hidden layers and 100 neurons in each layer is constructed and trained by using this dataset. Various types of validation of the ANN model are performed. It is found that the proposed ANN based model can effectively predict the frictional coefficient and cohesion of porous cement paste at the microscopic scale with a very good accuracy.

36 MATERIALS SCIENCE↗

2025 ASMS Investigation of the Collision-Induced Dissociation Mechanism of Protonated TODGA with IRIS

Title (20 words): Investigation of the Collision-Induced Dissociation Mechanism of Protonated TODGA with IRIS Introduction (120 words): One of the challenges facing wide-spread adoption of nuclear power is the development of efficient separation processes for used nuclear fuel. The molecules in separation processes are subjected to an extreme environment due to the high radiation fields from the used fuel and highly acidic media used for fuel dissolution, which results in significant molecular degradation, leading to reduced process efficiency. These degradation products must be identified and studied so mitigation strategies can be developed to maintain process efficiency. However, complex systems can have many degradation products, complicating identification. Untargeted analysis tools could be used to understand radiation chemistry in complex systems. However, this would necessitate improved understanding of the gas-phase fragmentation mechanisms of fuel cycle molecules like tetraoctyldiglycolamide (TODGA). Methods (120 words): The gas-phase fragmentation of protonated TODGA was investigated using collision-induced dissociation (CID), resonance ejection, and infrared ion spectroscopy (IRIS). CID and resonance ejection experiments were conducted using a Bruker Daltonics (Bremen, Gemany) SolariX XR fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer. IRIS spectra of protonated TODGA and its two CID fragmentation products were measured using a modified Bruker amaZon Speed ETD 3D quadrupole ion trap mass spectrometer coupled to the Free Electron Lasers for Infrared eXperiments (FELIX) free electron laser. Measured spectra were compared with density functional theory (DFT) calculations using the Gaussian 16, Revision C.02 software package with the ?B97X-D functional and def2-TZVPP basis sets. Candidate structures were generated using the CREST 3.0 conformational sampling software tool. Preliminary Data (300 words): Collision-induced dissociation of protonated TODGA ([C36H73N2O3]+, m/z=581.562) results two fragment ions, one at m/z=340.285 assigned as [C20H38NO3]+ and the other at m/z=312.290, assigned as [C19H38NO2]+. Based on the assigned formula and the structure of protonated TODGA, the fragment at m/z=340.285 is likely formed from elimination of neutral dioctylamine. Comparison of the IRIS spectrum of m/z=340.285 with DFT predictions suggests it contains a ring structure, and is assigned as N-octyl-N-(6-oxo-1,4-dioxan-2-ylidene)octan-1-aminium. Based on this structure and the structure of protonated TODGA, we hypothesize this fragment formed from elimination of neutral dioctylamine followed by a ring closure mechanism. Comparison of the IRIS spectrum of the fragment at m/z=312.290 with DFT predictions also indicated the presence of a ring structure, assigned as N-(1,3-dioxolan-4-ylidene)-N-octyloctan-1-aminium. This product could be formed from elimination of carbon monoxide from the ring of m/z=340.285 as a sequential fragmentation or formed directly from protonated TODGA via elimination of neutral N,N-dioctylformamide followed by a ring closure. Resonance ejection experiments where m/z=340 was continuously ejected from the IRC cell showed no decrease in intensity of m/z=312.290 across several collision energies, suggesting that the later, direct formation mechanism, dominates. The location of the ionizing proton in protonated TODGA is important for modeling the fragmentation mechanisms. DFT calculations suggested that the position of bands involving the coupled vibrations of the amide C—N and C=O bonds in TODGA are the most sensitive to proton location. Evaluation of the IRIS spectrum of protonated TODGA suggests that the ionizing proton is located between the two amid oxygens. This protonation location was calculated to lie approximately 30 kJ/mol lower in energy than the next lowest energy location, with the proton located solely on one of the amide oxygens. Novel aspect (20 words): Infrared ion spectroscopy combined with resonance ejection experiments and density functional theory to probe the collision-induced dissociation mechanism of tetraoctyldiglycolamide.

37 - INORGANIC, ORGANIC, PHYSICAL AND ANALYTICAL C↗

Matrix Metalloproteinases as Candidate Antigenic Determinants for Anti‐Tumor Autoantibodies in Human Ovarian Cancer: A Post Hoc Analysis

Circulating antibodies in patients with cancer can facilitate the identification of accessible epitopes on autoantigens expressed by tumors. To identify previously unrecognized protein targets in ovarian cancer, we computationally assessed a heptapeptide consensus motif (VPELGHE, flanked by two cysteine residues yielding a cyclic nonapeptide under oxidizing conditions) previously discovered via phage display-based epitope mapping of autoantibodies in patients. Eight proteins associated with ovarian cancer encompass amino acid sequences similar to the consensus motif and were, therefore, considered as candidate native autoantigens. Among these candidate targets, however, matrix metalloproteinase 14 (MMP14) demonstrates gene expression that is both high and negatively correlated with survival in ovarian cancer patient cohorts. MMP14 protein levels are also stable in tumor versus non-tumor tissues. Moreover, the corresponding heptapeptide mimic in MMP14 occurs within an α-helical secondary structural element observed in its catalytic domain. These findings demonstrate that a subset of patient-derived autoantibodies may interact with a previously unknown antigenic epitope found in MMP14 and other MMPs, thereby providing opportunities for the development of new targeted agents.

Biochemistry & Molecular Biology↗

TU-EF-BRA-00: MR Basics I

NMR, and Proton Density MRI of the 1D Patient - Anthony Wolbarst Net Voxel Magnetization, m(x,t). T1-MRI; The MRI Device - Lisa Lemen ‘Classical’ NMR; FID Imaging in 1D via k-Space - Nathan Yanasak Spin-Echo; S-E/Spin Warp in a 2D Slice - Ronald Price Magnetic resonance imaging not only reveals the structural, anatomic details of the body, as does CT, but also it can provide information on the physiological status and pathologies of its tissues, like nuclear medicine. It can display high-quality slice and 3D images of organs and vessels viewed from any perspective, with resolution better than 1 mm. MRI is perhaps most extraordinary and notable for the plethora of ways in which it can create unique forms of image contrast, reflective of fundamentally different biophysical phenomena. As with ultrasound, there is no risk from ionizing radiation to the patient or staff, since no X-rays or radioactive nuclei are involved. Instead, MRI harnesses magnetic fields and radio waves to probe the stable nuclei of the ordinary hydrogen atoms (isolated protons) occurring in water and lipid molecules within and around cells. MRI consists, in essence, of creating spatial maps of the electromagnetic environments around these hydrogen nuclei. Spatial variations in the proton milieus can be related to clinical differences in the biochemical and physiological properties and conditions of the associated tissues. Imaging of proton density (PD), and of the tissue proton spin relaxation times known as T1 and T2, all can reveal important clinical information, but they do so with approaches so dissimilar from one another that each is chosen for only certain clinical situations. T1 and T2 in a voxel are determined by different aspects of the rotations and other motions of the water and lipid molecules involved, as constrained by the local biophysical surroundings within and between its cells – and they, in turn, depend on the type of tissue and its state of health. Three other common applications of MRI exploit its capability to detect and image distinct movements of fluids: MR angiography (MRA), which rivals CT angiography but often requires no contrast medium, monitors the bulk flow of blood; functional MRI (f MRI), distinguishes the perfusion of oxygenated blood from that of de-oxygenated, and lights up parts of the brain that are activated by a stimulus, rather like PET; and diffusion tensor imaging (DTI) indicates the diffusion of free water along tracts of axons, thereby bringing nerve trunks into view. There are variants on all of these themes, and on others as well. Magnetic Resonance Spectroscopy (MRS), for example, can perform non-invasive ‘virtual biopsies’ that allow identification of certain cancers and other lesions. And an MRI-guided needle biopsy can sample brain tissue from a region only millimeters in dimensions. MRI, however, involves deeper and more complex aspects of physics, technology, and biology than do most other imaging modalities, and it is widely considered to be correspondingly more difficult to learn. We could probably cover all of this rather comprehensively if we had 50 hours available rather than 2 ̶ but, to paraphrase a former Secretary of Defense, you tell your story in the time you have allotted. The four presenters and another physicist, Kevin King from GE, have combined their efforts to co-author a single slide show that describes essentials of MRI as simply as possible. It is obviously far from thorough, but hopefully it will succeed in explaining some of the basics in a simplified but still valid fashion; in providing a taste of the numerous capabilities and complexities of the modality; and in whetting your appetite to learn more. Part I. NMR, and Proton Density MRI of the 1D Patient (Wolbarst), begins with an introductory case study that illustrates a half dozen ways in which MRI provides valuable clinical information. It then explores the nuclear magnetic resonance (NMR) phenomenon, which underlies MRI. NMR can be introduced with either of two approaches. In the first, one thinks (loosely) of the nuclei of hydrogen atoms as (rotating and charged and therefore) magnetic objects, whose spin-axes tend to align in a strong external magnetic field, much like a compass needle. As with the Bohr atom, this spin-up/spin-down picture is a highly abridged version of the full quantum mechanical treatment, but still it leads to some useful, legitimate pictures of the NMR process occurring within a voxel: When RF photons of the correct (Larmor) frequency elevate protons in a fixed magnetic field out of their lower-energy spin state into the upper, the NMR phenomenon is indicated by the detectable absorption of RF power. With the addition of a linear gradient field along a multi-voxel, one-dimensional patient/phantom, as well, we can determine the water content of each compartment – an example of a real MRI study, albeit in 1D. Part I concludes with a discussion of the net magnetization at position x, m0(x), under conditions of dynamic thermal equilibrium, which leads into: Part II. Net Voxel Magnetization, m(x,t); T1-MRI; The MRI Device (Lemen), investigates the biophysics of the form of proton spin relaxation process characterized by the time T1. It then moves on to the creation of an MR image that displays the spatial variation in the values of this clinically relevant parameter, again in 1D. Finally, the design and workings of a clinical MRI machine are sketched, in preparation for: Part III. ‘Classical’ NMR; FID Imaging in 1D via k-Space (Yanasak) presents the second standard approach to NMR and MRI, the classical model. It focuses on the time dependence of the net nuclear magnetization, m(x,t), the overall magnetic field generated by the cohort of protons in the voxel at position x. Quite remarkably, this nuclear net magnetization itself acts in a strong magnetic field like a gyroscope in a gravitational field. This tack is better for explaining Free Induction Decay (FID), which involves a brief introduction to the Fourier transform and k-space. This leads to conventional Spin-Echo (S-E) reconstruction techniques for creating clinical images from raw data, and sets the stage for: Part IV. Spin-Echo; S-E / Spin Warp in a 2D Slice (Price) discusses application of the S- E sequence of radiofrequency pulses and gradient magnetic fields to the 1D patient. T2 is introduced but not explained. This Part also considers how to manipulate the image acquisition parameters so as to generate clinical pictures with contrast dominated by spatial variations in PD, T1, or T2. We conclude by demonstrating the spin-warp approach to imaging in 2D with a simple 2×2, 4-voxel example. Much of this material is presented in more detail in the chapter “MRI of the One-dimensional Patient, Part I”, in Advances in Medical Physics, Vol 5 (2014). Copies are on display at the Medical Physics Publishing booth. Learning Objectives: The participant will learn about the processes of NMR and T1 spin relaxation in a tissue voxel in a uniform magnetic field. The participant will learn about combining spin-up/spin-down NMR and T1 processes with a linear gradient to effect frequency-encoding of voxel spatial position. This approach can be used to create proton density and T1 MRI maps, respectively, of the contents of multi-voxel 1D phantoms. The participant will learn about how the ‘classical’ model of NMR it can generate Free Induction Decay (FID) images of 1D phantoms, which involves the use of the Fourier transform in k-space. This can lead simply into standard Spin-Echo images. The participant will learn about extending Spin-Echo imaging into 2 and more dimensions.

60 APPLIED LIFE SCIENCES↗

TU-EF-BRA-01: NMR and Proton Density MRI of the 1D Patient

NMR, and Proton Density MRI of the 1D Patient - Anthony Wolbarst Net Voxel Magnetization, m(x,t). T1-MRI; The MRI Device - Lisa Lemen ‘Classical’ NMR; FID Imaging in 1D via k-Space - Nathan Yanasak Spin-Echo; S-E/Spin Warp in a 2D Slice - Ronald Price Magnetic resonance imaging not only reveals the structural, anatomic details of the body, as does CT, but also it can provide information on the physiological status and pathologies of its tissues, like nuclear medicine. It can display high-quality slice and 3D images of organs and vessels viewed from any perspective, with resolution better than 1 mm. MRI is perhaps most extraordinary and notable for the plethora of ways in which it can create unique forms of image contrast, reflective of fundamentally different biophysical phenomena. As with ultrasound, there is no risk from ionizing radiation to the patient or staff, since no X-rays or radioactive nuclei are involved. Instead, MRI harnesses magnetic fields and radio waves to probe the stable nuclei of the ordinary hydrogen atoms (isolated protons) occurring in water and lipid molecules within and around cells. MRI consists, in essence, of creating spatial maps of the electromagnetic environments around these hydrogen nuclei. Spatial variations in the proton milieus can be related to clinical differences in the biochemical and physiological properties and conditions of the associated tissues. Imaging of proton density (PD), and of the tissue proton spin relaxation times known as T1 and T2, all can reveal important clinical information, but they do so with approaches so dissimilar from one another that each is chosen for only certain clinical situations. T1 and T2 in a voxel are determined by different aspects of the rotations and other motions of the water and lipid molecules involved, as constrained by the local biophysical surroundings within and between its cells – and they, in turn, depend on the type of tissue and its state of health. Three other common applications of MRI exploit its capability to detect and image distinct movements of fluids: MR angiography (MRA), which rivals CT angiography but often requires no contrast medium, monitors the bulk flow of blood; functional MRI ( f MRI), distinguishes the perfusion of oxygenated blood from that of de-oxygenated, and lights up parts of the brain that are activated by a stimulus, rather like PET; and diffusion tensor imaging (DTI) indicates the diffusion of free water along tracts of axons, thereby bringing nerve trunks into view. There are variants on all of these themes, and on others as well. Magnetic Resonance Spectroscopy (MRS), for example, can perform non-invasive ‘virtual biopsies’ that allow identification of certain cancers and other lesions. And an MRI-guided needle biopsy can sample brain tissue from a region only millimeters in dimensions. MRI, however, involves deeper and more complex aspects of physics, technology, and biology than do most other imaging modalities, and it is widely considered to be correspondingly more difficult to learn. We could probably cover all of this rather comprehensively if we had 50 hours available rather than 2 ̶ but, to paraphrase a former Secretary of Defense, you tell your story in the time you have allotted. The four presenters and another physicist, Kevin King from GE, have combined their efforts to co-author a single slide show that describes essentials of MRI as simply as possible. It is obviously far from thorough, but hopefully it will succeed in explaining some of the basics in a simplified but still valid fashion; in providing a taste of the numerous capabilities and complexities of the modality; and in whetting your appetite to learn more. Part I. NMR, and Proton Density MRI of the 1D Patient (Wolbarst), begins with an introductory case study that illustrates a half dozen ways in which MRI provides valuable clinical information. It then explores the nuclear magnetic resonance (NMR) phenomenon, which underlies MRI. NMR can be introduced with either of two approaches. In the first, one thinks (loosely) of the nuclei of hydrogen atoms as (rotating and charged and therefore) magnetic objects, whose spin-axes tend to align in a strong external magnetic field, much like a compass needle. As with the Bohr atom, this spin-up/spin-down picture is a highly abridged version of the full quantum mechanical treatment, but still it leads to some useful, legitimate pictures of the NMR process occurring within a voxel: When RF photons of the correct (Larmor) frequency elevate protons in a fixed magnetic field out of their lower-energy spin state into the upper, the NMR phenomenon is indicated by the detectable absorption of RF power. With the addition of a linear gradient field along a multi-voxel, one-dimensional patient/phantom, as well, we can determine the water content of each compartment – an example of a real MRI study, albeit in 1D. Part I concludes with a discussion of the net magnetization at position x, m0(x), under conditions of dynamic thermal equilibrium, which leads into: Part II. Net Voxel Magnetization, m(x,t); T1-MRI; The MRI Device (Lemen), investigates the biophysics of the form of proton spin relaxation process characterized by the time T1. It then moves on to the creation of an MR image that displays the spatial variation in the values of this clinically relevant parameter, again in 1D. Finally, the design and workings of a clinical MRI machine are sketched, in preparation for: Part III. ‘Classical’ NMR; FID Imaging in 1D via k-Space (Yanasak) presents the second standard approach to NMR and MRI, the classical model. It focuses on the time dependence of the net nuclear magnetization, m(x,t), the overall magnetic field generated by the cohort of protons in the voxel at position x. Quite remarkably, this nuclear net magnetization itself acts in a strong magnetic field like a gyroscope in a gravitational field. This tack is better for explaining Free Induction Decay (FID), which involves a brief introduction to the Fourier transform and k-space. This leads to conventional Spin-Echo (S-E) reconstruction techniques for creating clinical images from raw data, and sets the stage for: Part IV. Spin-Echo; S-E / Spin Warp in a 2D Slice (Price) discusses application of the S- E sequence of radiofrequency pulses and gradient magnetic fields to the 1D patient. T2 is introduced but not explained. This Part also considers how to manipulate the image acquisition parameters so as to generate clinical pictures with contrast dominated by spatial variations in PD, T1, or T2. We conclude by demonstrating the spin-warp approach to imaging in 2D with a simple 2×2, 4-voxel example. Much of this material is presented in more detail in the chapter “MRI of the One-dimensional Patient, Part I”, in Advances in Medical Physics, Vol 5 (2014). Copies are on display at the Medical Physics Publishing booth. Learning Objectives: The participant will learn about the processes of NMR and T1 spin relaxation in a tissue voxel in a uniform magnetic field. The participant will learn about combining spin-up/spin-down NMR and T1 processes with a linear gradient to effect frequency-encoding of voxel spatial position. This approach can be used to create proton density and T1 MRI maps, respectively, of the contents of multi-voxel 1D phantoms. The participant will learn about how the ‘classical’ model of NMR it can generate Free Induction Decay (FID) images of 1D phantoms, which involves the use of the Fourier transform in k-space. This can lead simply into standard Spin-Echo images. The participant will learn about extending Spin-Echo imaging into 2 and more dimensions.

60 APPLIED LIFE SCIENCES↗

TU-EF-BRA-02: Longitudinal Proton Spin Relaxation and T1-Imaging

NMR, and Proton Density MRI of the 1D Patient - Anthony Wolbarst Net Voxel Magnetization, m(x,t). T1-MRI; The MRI Device - Lisa Lemen ‘Classical’ NMR; FID Imaging in 1D via k-Space - Nathan Yanasak Spin-Echo; S-E/Spin Warp in a 2D Slice - Ronald Price Magnetic resonance imaging not only reveals the structural, anatomic details of the body, as does CT, but also it can provide information on the physiological status and pathologies of its tissues, like nuclear medicine. It can display high-quality slice and 3D images of organs and vessels viewed from any perspective, with resolution better than 1 mm. MRI is perhaps most extraordinary and notable for the plethora of ways in which it can create unique forms of image contrast, reflective of fundamentally different biophysical phenomena. As with ultrasound, there is no risk from ionizing radiation to the patient or staff, since no X-rays or radioactive nuclei are involved. Instead, MRI harnesses magnetic fields and radio waves to probe the stable nuclei of the ordinary hydrogen atoms (isolated protons) occurring in water and lipid molecules within and around cells. MRI consists, in essence, of creating spatial maps of the electromagnetic environments around these hydrogen nuclei. Spatial variations in the proton milieus can be related to clinical differences in the biochemical and physiological properties and conditions of the associated tissues. Imaging of proton density (PD), and of the tissue proton spin relaxation times known as T1 and T2, all can reveal important clinical information, but they do so with approaches so dissimilar from one another that each is chosen for only certain clinical situations. T1 and T2 in a voxel are determined by different aspects of the rotations and other motions of the water and lipid molecules involved, as constrained by the local biophysical surroundings within and between its cells – and they, in turn, depend on the type of tissue and its state of health. Three other common applications of MRI exploit its capability to detect and image distinct movements of fluids: MR angiography (MRA), which rivals CT angiography but often requires no contrast medium, monitors the bulk flow of blood; functional MRI ( f MRI), distinguishes the perfusion of oxygenated blood from that of de-oxygenated, and lights up parts of the brain that are activated by a stimulus, rather like PET; and diffusion tensor imaging (DTI) indicates the diffusion of free water along tracts of axons, thereby bringing nerve trunks into view. There are variants on all of these themes, and on others as well. Magnetic Resonance Spectroscopy (MRS), for example, can perform non-invasive ‘virtual biopsies’ that allow identification of certain cancers and other lesions. And an MRI-guided needle biopsy can sample brain tissue from a region only millimeters in dimensions. MRI, however, involves deeper and more complex aspects of physics, technology, and biology than do most other imaging modalities, and it is widely considered to be correspondingly more difficult to learn. We could probably cover all of this rather comprehensively if we had 50 hours available rather than 2 ̶ but, to paraphrase a former Secretary of Defense, you tell your story in the time you have allotted. The four presenters and another physicist, Kevin King from GE, have combined their efforts to co-author a single slide show that describes essentials of MRI as simply as possible. It is obviously far from thorough, but hopefully it will succeed in explaining some of the basics in a simplified but still valid fashion; in providing a taste of the numerous capabilities and complexities of the modality; and in whetting your appetite to learn more. Part I. NMR, and Proton Density MRI of the 1D Patient (Wolbarst), begins with an introductory case study that illustrates a half dozen ways in which MRI provides valuable clinical information. It then explores the nuclear magnetic resonance (NMR) phenomenon, which underlies MRI. NMR can be introduced with either of two approaches. In the first, one thinks (loosely) of the nuclei of hydrogen atoms as (rotating and charged and therefore) magnetic objects, whose spin-axes tend to align in a strong external magnetic field, much like a compass needle. As with the Bohr atom, this spin-up/spin-down picture is a highly abridged version of the full quantum mechanical treatment, but still it leads to some useful, legitimate pictures of the NMR process occurring within a voxel: When RF photons of the correct (Larmor) frequency elevate protons in a fixed magnetic field out of their lower-energy spin state into the upper, the NMR phenomenon is indicated by the detectable absorption of RF power. With the addition of a linear gradient field along a multi-voxel, one-dimensional patient/phantom, as well, we can determine the water content of each compartment – an example of a real MRI study, albeit in 1D. Part I concludes with a discussion of the net magnetization at position x, m0(x), under conditions of dynamic thermal equilibrium, which leads into: Part II. Net Voxel Magnetization, m(x,t); T1-MRI; The MRI Device (Lemen), investigates the biophysics of the form of proton spin relaxation process characterized by the time T1. It then moves on to the creation of an MR image that displays the spatial variation in the values of this clinically relevant parameter, again in 1D. Finally, the design and workings of a clinical MRI machine are sketched, in preparation for: Part III. ‘Classical’ NMR; FID Imaging in 1D via k-Space (Yanasak) presents the second standard approach to NMR and MRI, the classical model. It focuses on the time dependence of the net nuclear magnetization, m(x,t), the overall magnetic field generated by the cohort of protons in the voxel at position x. Quite remarkably, this nuclear net magnetization itself acts in a strong magnetic field like a gyroscope in a gravitational field. This tack is better for explaining Free Induction Decay (FID), which involves a brief introduction to the Fourier transform and k-space. This leads to conventional Spin-Echo (S-E) reconstruction techniques for creating clinical images from raw data, and sets the stage for: Part IV. Spin-Echo; S-E / Spin Warp in a 2D Slice (Price) discusses application of the S- E sequence of radiofrequency pulses and gradient magnetic fields to the 1D patient. T2 is introduced but not explained. This Part also considers how to manipulate the image acquisition parameters so as to generate clinical pictures with contrast dominated by spatial variations in PD, T1, or T2. We conclude by demonstrating the spin-warp approach to imaging in 2D with a simple 2×2, 4-voxel example. Much of this material is presented in more detail in the chapter “MRI of the One-dimensional Patient, Part I”, in Advances in Medical Physics, Vol 5 (2014). Copies are on display at the Medical Physics Publishing booth. Learning Objectives: The participant will learn about the processes of NMR and T1 spin relaxation in a tissue voxel in a uniform magnetic field. The participant will learn about combining spin-up/spin-down NMR and T1 processes with a linear gradient to effect frequency-encoding of voxel spatial position. This approach can be used to create proton density and T1 MRI maps, respectively, of the contents of multi-voxel 1D phantoms. The participant will learn about how the ‘classical’ model of NMR it can generate Free Induction Decay (FID) images of 1D phantoms, which involves the use of the Fourier transform in k-space. This can lead simply into standard Spin-Echo images. The participant will learn about extending Spin-Echo imaging into 2 and more dimensions.

60 APPLIED LIFE SCIENCES↗

TU-EF-BRA-03: Free Induction Decay (without the Decay) and Spin-Echo Imaging

NMR, and Proton Density MRI of the 1D Patient - Anthony Wolbarst Net Voxel Magnetization, m(x,t). T1-MRI; The MRI Device - Lisa Lemen ‘Classical’ NMR; FID Imaging in 1D via k-Space - Nathan Yanasak Spin-Echo; S-E/Spin Warp in a 2D Slice - Ronald Price Magnetic resonance imaging not only reveals the structural, anatomic details of the body, as does CT, but also it can provide information on the physiological status and pathologies of its tissues, like nuclear medicine. It can display high-quality slice and 3D images of organs and vessels viewed from any perspective, with resolution better than 1 mm. MRI is perhaps most extraordinary and notable for the plethora of ways in which it can create unique forms of image contrast, reflective of fundamentally different biophysical phenomena. As with ultrasound, there is no risk from ionizing radiation to the patient or staff, since no X-rays or radioactive nuclei are involved. Instead, MRI harnesses magnetic fields and radio waves to probe the stable nuclei of the ordinary hydrogen atoms (isolated protons) occurring in water and lipid molecules within and around cells. MRI consists, in essence, of creating spatial maps of the electromagnetic environments around these hydrogen nuclei. Spatial variations in the proton milieus can be related to clinical differences in the biochemical and physiological properties and conditions of the associated tissues. Imaging of proton density (PD), and of the tissue proton spin relaxation times known as T1 and T2, all can reveal important clinical information, but they do so with approaches so dissimilar from one another that each is chosen for only certain clinical situations. T1 and T2 in a voxel are determined by different aspects of the rotations and other motions of the water and lipid molecules involved, as constrained by the local biophysical surroundings within and between its cells – and they, in turn, depend on the type of tissue and its state of health. Three other common applications of MRI exploit its capability to detect and image distinct movements of fluids: MR angiography (MRA), which rivals CT angiography but often requires no contrast medium, monitors the bulk flow of blood; functional MRI ( f MRI), distinguishes the perfusion of oxygenated blood from that of de-oxygenated, and lights up parts of the brain that are activated by a stimulus, rather like PET; and diffusion tensor imaging (DTI) indicates the diffusion of free water along tracts of axons, thereby bringing nerve trunks into view. There are variants on all of these themes, and on others as well. Magnetic Resonance Spectroscopy (MRS), for example, can perform non-invasive ‘virtual biopsies’ that allow identification of certain cancers and other lesions. And an MRI-guided needle biopsy can sample brain tissue from a region only millimeters in dimensions. MRI, however, involves deeper and more complex aspects of physics, technology, and biology than do most other imaging modalities, and it is widely considered to be correspondingly more difficult to learn. We could probably cover all of this rather comprehensively if we had 50 hours available rather than 2 ̶ but, to paraphrase a former Secretary of Defense, you tell your story in the time you have allotted. The four presenters and another physicist, Kevin King from GE, have combined their efforts to co-author a single slide show that describes essentials of MRI as simply as possible. It is obviously far from thorough, but hopefully it will succeed in explaining some of the basics in a simplified but still valid fashion; in providing a taste of the numerous capabilities and complexities of the modality; and in whetting your appetite to learn more. Part I. NMR, and Proton Density MRI of the 1D Patient (Wolbarst), begins with an introductory case study that illustrates a half dozen ways in which MRI provides valuable clinical information. It then explores the nuclear magnetic resonance (NMR) phenomenon, which underlies MRI. NMR can be introduced with either of two approaches. In the first, one thinks (loosely) of the nuclei of hydrogen atoms as (rotating and charged and therefore) magnetic objects, whose spin-axes tend to align in a strong external magnetic field, much like a compass needle. As with the Bohr atom, this spin-up/spin-down picture is a highly abridged version of the full quantum mechanical treatment, but still it leads to some useful, legitimate pictures of the NMR process occurring within a voxel: When RF photons of the correct (Larmor) frequency elevate protons in a fixed magnetic field out of their lower-energy spin state into the upper, the NMR phenomenon is indicated by the detectable absorption of RF power. With the addition of a linear gradient field along a multi-voxel, one-dimensional patient/phantom, as well, we can determine the water content of each compartment – an example of a real MRI study, albeit in 1D. Part I concludes with a discussion of the net magnetization at position x, m0(x), under conditions of dynamic thermal equilibrium, which leads into: Part II. Net Voxel Magnetization, m(x,t); T1-MRI; The MRI Device (Lemen), investigates the biophysics of the form of proton spin relaxation process characterized by the time T1. It then moves on to the creation of an MR image that displays the spatial variation in the values of this clinically relevant parameter, again in 1D. Finally, the design and workings of a clinical MRI machine are sketched, in preparation for: Part III. ‘Classical’ NMR; FID Imaging in 1D via k-Space (Yanasak) presents the second standard approach to NMR and MRI, the classical model. It focuses on the time dependence of the net nuclear magnetization, m(x,t), the overall magnetic field generated by the cohort of protons in the voxel at position x. Quite remarkably, this nuclear net magnetization itself acts in a strong magnetic field like a gyroscope in a gravitational field. This tack is better for explaining Free Induction Decay (FID), which involves a brief introduction to the Fourier transform and k-space. This leads to conventional Spin-Echo (S-E) reconstruction techniques for creating clinical images from raw data, and sets the stage for: Part IV. Spin-Echo; S-E / Spin Warp in a 2D Slice (Price) discusses application of the S- E sequence of radiofrequency pulses and gradient magnetic fields to the 1D patient. T2 is introduced but not explained. This Part also considers how to manipulate the image acquisition parameters so as to generate clinical pictures with contrast dominated by spatial variations in PD, T1, or T2. We conclude by demonstrating the spin-warp approach to imaging in 2D with a simple 2×2, 4-voxel example. Much of this material is presented in more detail in the chapter “MRI of the One-dimensional Patient, Part I”, in Advances in Medical Physics, Vol 5 (2014). Copies are on display at the Medical Physics Publishing booth. Learning Objectives: The participant will learn about the processes of NMR and T1 spin relaxation in a tissue voxel in a uniform magnetic field. The participant will learn about combining spin-up/spin-down NMR and T1 processes with a linear gradient to effect frequency-encoding of voxel spatial position. This approach can be used to create proton density and T1 MRI maps, respectively, of the contents of multi-voxel 1D phantoms. The participant will learn about how the ‘classical’ model of NMR it can generate Free Induction Decay (FID) images of 1D phantoms, which involves the use of the Fourier transform in k-space. This can lead simply into standard Spin-Echo images. The participant will learn about extending Spin-Echo imaging into 2 and more dimensions.

60 APPLIED LIFE SCIENCES↗

TU-EF-BRA-04: Into 2, 3, and 4 Dimensions

NMR, and Proton Density MRI of the 1D Patient - Anthony Wolbarst Net Voxel Magnetization, m(x,t). T1-MRI; The MRI Device - Lisa Lemen ‘Classical’ NMR; FID Imaging in 1D via k-Space - Nathan Yanasak Spin-Echo; S-E/Spin Warp in a 2D Slice - Ronald Price Magnetic resonance imaging not only reveals the structural, anatomic details of the body, as does CT, but also it can provide information on the physiological status and pathologies of its tissues, like nuclear medicine. It can display high-quality slice and 3D images of organs and vessels viewed from any perspective, with resolution better than 1 mm. MRI is perhaps most extraordinary and notable for the plethora of ways in which it can create unique forms of image contrast, reflective of fundamentally different biophysical phenomena. As with ultrasound, there is no risk from ionizing radiation to the patient or staff, since no X-rays or radioactive nuclei are involved. Instead, MRI harnesses magnetic fields and radio waves to probe the stable nuclei of the ordinary hydrogen atoms (isolated protons) occurring in water and lipid molecules within and around cells. MRI consists, in essence, of creating spatial maps of the electromagnetic environments around these hydrogen nuclei. Spatial variations in the proton milieus can be related to clinical differences in the biochemical and physiological properties and conditions of the associated tissues. Imaging of proton density (PD), and of the tissue proton spin relaxation times known as T1 and T2, all can reveal important clinical information, but they do so with approaches so dissimilar from one another that each is chosen for only certain clinical situations. T1 and T2 in a voxel are determined by different aspects of the rotations and other motions of the water and lipid molecules involved, as constrained by the local biophysical surroundings within and between its cells – and they, in turn, depend on the type of tissue and its state of health. Three other common applications of MRI exploit its capability to detect and image distinct movements of fluids: MR angiography (MRA), which rivals CT angiography but often requires no contrast medium, monitors the bulk flow of blood; functional MRI ( f MRI), distinguishes the perfusion of oxygenated blood from that of de-oxygenated, and lights up parts of the brain that are activated by a stimulus, rather like PET; and diffusion tensor imaging (DTI) indicates the diffusion of free water along tracts of axons, thereby bringing nerve trunks into view. There are variants on all of these themes, and on others as well. Magnetic Resonance Spectroscopy (MRS), for example, can perform non-invasive ‘virtual biopsies’ that allow identification of certain cancers and other lesions. And an MRI-guided needle biopsy can sample brain tissue from a region only millimeters in dimensions. MRI, however, involves deeper and more complex aspects of physics, technology, and biology than do most other imaging modalities, and it is widely considered to be correspondingly more difficult to learn. We could probably cover all of this rather comprehensively if we had 50 hours available rather than 2 ̶ but, to paraphrase a former Secretary of Defense, you tell your story in the time you have allotted. The four presenters and another physicist, Kevin King from GE, have combined their efforts to co-author a single slide show that describes essentials of MRI as simply as possible. It is obviously far from thorough, but hopefully it will succeed in explaining some of the basics in a simplified but still valid fashion; in providing a taste of the numerous capabilities and complexities of the modality; and in whetting your appetite to learn more. Part I. NMR, and Proton Density MRI of the 1D Patient (Wolbarst), begins with an introductory case study that illustrates a half dozen ways in which MRI provides valuable clinical information. It then explores the nuclear magnetic resonance (NMR) phenomenon, which underlies MRI. NMR can be introduced with either of two approaches. In the first, one thinks (loosely) of the nuclei of hydrogen atoms as (rotating and charged and therefore) magnetic objects, whose spin-axes tend to align in a strong external magnetic field, much like a compass needle. As with the Bohr atom, this spin-up/spin-down picture is a highly abridged version of the full quantum mechanical treatment, but still it leads to some useful, legitimate pictures of the NMR process occurring within a voxel: When RF photons of the correct (Larmor) frequency elevate protons in a fixed magnetic field out of their lower-energy spin state into the upper, the NMR phenomenon is indicated by the detectable absorption of RF power. With the addition of a linear gradient field along a multi-voxel, one-dimensional patient/phantom, as well, we can determine the water content of each compartment – an example of a real MRI study, albeit in 1D. Part I concludes with a discussion of the net magnetization at position x, m0(x), under conditions of dynamic thermal equilibrium, which leads into: Part II. Net Voxel Magnetization, m(x,t); T1-MRI; The MRI Device (Lemen), investigates the biophysics of the form of proton spin relaxation process characterized by the time T1. It then moves on to the creation of an MR image that displays the spatial variation in the values of this clinically relevant parameter, again in 1D. Finally, the design and workings of a clinical MRI machine are sketched, in preparation for: Part III. ‘Classical’ NMR; FID Imaging in 1D via k-Space (Yanasak) presents the second standard approach to NMR and MRI, the classical model. It focuses on the time dependence of the net nuclear magnetization, m(x,t), the overall magnetic field generated by the cohort of protons in the voxel at position x. Quite remarkably, this nuclear net magnetization itself acts in a strong magnetic field like a gyroscope in a gravitational field. This tack is better for explaining Free Induction Decay (FID), which involves a brief introduction to the Fourier transform and k-space. This leads to conventional Spin-Echo (S-E) reconstruction techniques for creating clinical images from raw data, and sets the stage for: Part IV. Spin-Echo; S-E / Spin Warp in a 2D Slice (Price) discusses application of the S- E sequence of radiofrequency pulses and gradient magnetic fields to the 1D patient. T2 is introduced but not explained. This Part also considers how to manipulate the image acquisition parameters so as to generate clinical pictures with contrast dominated by spatial variations in PD, T1, or T2. We conclude by demonstrating the spin-warp approach to imaging in 2D with a simple 2×2, 4-voxel example. Much of this material is presented in more detail in the chapter “MRI of the One-dimensional Patient, Part I”, in Advances in Medical Physics, Vol 5 (2014). Copies are on display at the Medical Physics Publishing booth. Learning Objectives: The participant will learn about the processes of NMR and T1 spin relaxation in a tissue voxel in a uniform magnetic field. The participant will learn about combining spin-up/spin-down NMR and T1 processes with a linear gradient to effect frequency-encoding of voxel spatial position. This approach can be used to create proton density and T1 MRI maps, respectively, of the contents of multi-voxel 1D phantoms. The participant will learn about how the ‘classical’ model of NMR it can generate Free Induction Decay (FID) images of 1D phantoms, which involves the use of the Fourier transform in k-space. This can lead simply into standard Spin-Echo images. The participant will learn about extending Spin-Echo imaging into 2 and more dimensions.

60 APPLIED LIFE SCIENCES↗

Association Mapping of Cell Wall Synthesis Regulatory Genes and Cell Wall Quality in Switchgrass

Inefficient conversion of biomass to biofuels is one of the main barriers for biofuel production from such materials. Approximately half of polysaccharides in biomass remain unused by typical biochemical conversion methods. Conversion efficiency is influenced by the composition and structure of cell walls of biomass. Grasses such as wheat, maize, and rice, as well as dedicated perennial bioenergy crops, like switchgrass, make up ~55% of biomass that can be produced in the United States. Grass cell walls have a different composition and patterning compared with dicotyledonous plants, including the well-studied model plant, Arabidopsis. This project identified genetic determinants of cell wall composition in grasses using both naturally occurring genetic variation of switchgrass and gene network reconstruction and functional assays in rice. In addition, the project linked functional data in rice and other species to switchgrass improvement efforts through curation of the most abundant class of regulators in the switchgrass genome. Characterizing natural diversity of switchgrass for variation in cell wall composition and properties, also known as quality, provides an unbiased avenue for identifying biologically viable diversity in switchgrass cell walls. To characterizing natural diversity, this project generated cell wall composition and enzymatic deconstruction data for ~450 genotypes of the Switchgrass Southern Association Collection (SSAC), a diverse collection composed of 36 switchgrass accessions from the southern U.S. distribution of switchgrass. Comparing these data with other measures of cell wall quality for the same samples demonstrated the complementary nature of the diverse characterization platforms now being used for biomass characterization. Association of the composition data with ~3.2K single nucleotide variant markers identified six significant single nucleotide variant markers co-associated with digestibility and another compositional trait. These markers might be used to select switchgrass genotypes with improved composition in breeding programs for biofuel and forage production. Because the SSAC continues to be characterized by collaborators in the bioenergy community, the data generated will be used to identify additional markers in higher resolution genotyping data to approach identifying the genes and alleles that cause natural variation in switchgrass cell wall quality. For example, these markers can be surveyed in the 2100-member Oklahoma Southern and Northern Lowland switchgrass collections that this project also characterized. An orthogonal approach to biodiversity studies, using comparative functional genomics permits systematic querying of how much regulatory information is likely to be transferable from dicots to grasses and use of accumulated functional genomics resources for better-characterized grass species, such as rice, itself a biomass source in global agriculture and in certain regions. The project generated and tested a number of specific hypotheses regarding cell wall transcription factors and enzymes of grasses. To aid identification of cell wall regulators, the project assembled a novel, highdepth and -quality gene association network using a general linearized model scoring system to combine rice gene network data. Using known or putative orthologs of Arabidopsis cell wall biosynthesis genes and regulators, the project pulled from this network a cell wall sub-network that includes 96 transcription factors. Reverse genetics of a co-ortholog of the Arabidopsis MYB61 transcription factor in rice revealed that this regulatory node has evolved the ability to regulate grass-specific cell wall synthesis enzymes. A transcription factor with such activity has not been previously characterized to our knowledge, representing a major conclusion of this work. Changes in gene expression in a protoplast-based assay demonstrated positive or negative roles in cell wall regulation for eleven other transcription factors from the rice gene network. Eight of fifteen (53%) of these have not previously been examined for this function. Some of these may represent novel grass-diverged cell wall regulators, while others are likely to have this function across angiosperms. A parallel effort of this project to expand knowledge of enzymes that have evolved to function in grass cell wall synthesis, revealed that a grass-diverged enzyme in rice, OsAT 5 , ferulates monolignols that are naturally incorporated into grass cell walls. This finding opens potential natural selection avenues for improving biomass composition for downstream processing by weak base pretreatment. Thus, this project has significantly expanded knowledge of cell wall synthesis and regulation in rice, information that can be used in reverse genetics and synthetic biology approaches to re-engineer cell walls for improved production of biofuel and high-value products. To lay the foundation for translating these results directly for switchgrass improvement, the project employed a comparative phylogenetic analysis of the major group of cell wall transcription factors that have been found to function in cell wall regulation, the R 2 R 3 MYBs. This analysis concluded that known cell wall regulators are largely conserved across switchgrass, rice, maize, poplar, and Arabidopsis. This interpretation is also largely consistent with the gene network analysis described above, though both approaches provide evidence that some co-orthologs of Arabidopsis regulators have diminished or increased in importance based on gene expression patterns. Also, several clades containing dicot cell wall regulators have expanded, consistent with the evolution of new cell wall regulators. This latter result is supported by functional analysis of the R 2 R 3 MYB protein SWAM 1 in a collaboration between this project and the DOE-funded group of Dr. S. Hazen at the University of Massachusettes. The curation of the switchgrass genome through this project provides specific targets for future engineering of switchgrass cell wall regulation and may also facilitate identification of regulators that underlie the molecular markers that are genetically linked to differences in cell wall quality. With the goal of spurring further research and technological developments in lignocellulosic biofuel production, this work has been communicated to the bioenergy and cell wall communities though various presentations and publications. To date, three manuscripts have been published, two others are near to publication, three others are in an advanced state, and two to four more are likely to be written based on analyses still in progress. In addition, project participants have presented thirteen posters and talks at regional, national, and international meetings about aspects of this project. In sum, the work supported by this funding has made and communicated significant progress in identifying the genes that grasses use for cell wall synthesis and regulation, information that will be used by project participants and others to improve the efficiency of conversion of lignocellulosic biomass to biofuels.

09 BIOMASS FUELS↗

Interplay between Mixed and Pure Exciton States Controls Singlet Fission in Rubrene Single Crystals

Singlet fission (SF) is a multielectron process in which one singlet exciton S converts into a pair of separated triplet excitons T. SF is widely studied as it may help overcome the Shockley−Queisser efficiency limit for semiconductor photovoltaic cells. To elucidate and control the SF mechanism, great attention has been given to the identification of intermediate states in SF materials, which often appear elusive due to the complexity and fast time scales of the SF process. Here, we apply 14 fs-1 ms transient absorption techniques to high-purity rubrene single crystals to disentangle the intrinsic fission dynamics from the effects of defects and grain boundaries and to identify reliably the fission intermediates. Our data demonstrates that above-gap excitation directly generates a hybrid vibronically assisted mixture of singlet state and triplet-pair multiexciton [S/TT], which rapidly (<100 fs) and coherently branches into pure singlet or triplet excitations. The relaxation of [S/TT] to S is followed by a relatively slow and temperature-activated (48 meV activation energy) incoherent fission process. The SF competing pathways and intermediates revealed here unify the observations and models presented in previous studies of SF in rubrene and offer alternative strategies for the development of SF-enhanced photovoltaic materials.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Extracellular calcium sensing and extracellular calcium signaling

The cloning of a G protein-coupled extracellular Ca(2+) (Ca(o)(2+))-sensing receptor (CaR) has elucidated the molecular basis for many of the previously recognized effects of Ca(o)(2+) on tissues that maintain systemic Ca(o)(2+) homeostasis, especially parathyroid chief cells and several cells in the kidney. The availability of the cloned CaR enabled the development of DNA and antibody probes for identifying the CaR's mRNA and protein, respectively, within these and other tissues. It also permitted the identification of human diseases resulting from inactivating or activating mutations of the CaR gene and the subsequent generation of mice with targeted disruption of the CaR gene. The characteristic alterations in parathyroid and renal function in these patients and in the mice with "knockout" of the CaR gene have provided valuable information on the CaR's physiological roles in these tissues participating in mineral ion homeostasis. Nevertheless, relatively little is known about how the CaR regulates other tissues involved in systemic Ca(o)(2+) homeostasis, particularly bone and intestine. Moreover, there is evidence that additional Ca(o)(2+) sensors may exist in bone cells that mediate some or even all of the known effects of Ca(o)(2+) on these cells. Even more remains to be learned about the CaR's function in the rapidly growing list of cells that express it but are uninvolved in systemic Ca(o)(2+) metabolism. Available data suggest that the receptor serves numerous roles outside of systemic mineral ion homeostasis, ranging from the regulation of hormonal secretion and the activities of various ion channels to the longer term control of gene expression, programmed cell death (apoptosis), and cellular proliferation. In some cases, the CaR on these "nonhomeostatic" cells responds to local changes in Ca(o)(2+) taking place within compartments of the extracellular fluid (ECF) that communicate with the outside environment (e.g., the gastrointestinal tract). In others, localized changes in Ca(o)(2+) within the ECF can originate from several mechanisms, including fluxes of calcium ions into or out of cellular or extracellular stores or across epithelium that absorb or secrete Ca(2+). In any event, the CaR and other receptors/sensors for Ca(o)(2+) and probably for other extracellular ions represent versatile regulators of numerous cellular functions and may serve as important therapeutic targets.

Review↗