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At least 1,315 records · Page 73

Alpha‐ and beta‐mannan utilization by marine Bacteroidetes

Summary Marine microscopic algae carry out about half of the global carbon dioxide fixation into organic matter. They provide organic substrates for marine microbes such as members of the Bacteroidetes that degrade algal polysaccharides using carbohydrate‐active enzymes (CAZymes). In Bacteroidetes genomes CAZyme encoding genes are mostly grouped in distinct regions termed polysaccharide utilization loci (PULs). While some studies have shown involvement of PULs in the degradation of algal polysaccharides, the specific substrates are for the most part still unknown. We investigated four marine Bacteroidetes isolated from the southern North Sea that harbour putative mannan‐specific PULs. These PULs are similarly organized as PULs in human gut Bacteroides that digest α‐ and β‐mannans from yeasts and plants respectively. Using proteomics and defined growth experiments with polysaccharides as sole carbon sources we could show that the investigated marine Bacteroidetes express the predicted functional proteins required for α‐ and β‐mannan degradation. Our data suggest that algal mannans play an as yet unknown important role in the marine carbon cycle, and that biochemical principles established for gut or terrestrial microbes also apply to marine bacteria, even though their PULs are evolutionarily distant.

Chen, Jing↗

GalR, GalX and AraR co-regulate $\tiny {D}$-galactose and $\tiny{L}$-arabinose utilization in Aspergillus nidulans

Filamentous fungi produce a wide variety of enzymes in order to efficiently degrade plant cell wall polysaccharides. The production of these enzymes is controlled by transcriptional regulators, which also control the catabolic pathways that convert the released monosaccharides. Two transcriptional regulators, GalX and GalR, control d-galactose utilization in the model filamentous fungus Aspergillus nidulans, while the arabinanolytic regulator AraR regulates l-arabinose catabolism. d-Galactose and l-arabinose are commonly found together in polysaccharides, such as arabinogalactan, xylan and rhamnogalacturonan I. Therefore, the catabolic pathways that convert d-galactose and l-arabinose are often also likely to be active simultaneously. In this study, we investigated the interaction between GalX, GalR and AraR in d-galactose and l-arabinose catabolism. For this, we generated single, double and triple mutants of the three regulators, and analysed their growth and enzyme and gene expression profiles. Our results clearly demonstrated that GalX, GalR and AraR co-regulate d-galactose catabolism in A. nidulans. GalX has a prominent role on the regulation of genes of d-galactose oxido-reductive pathway, while AraR can compensate for the absence of GalR and/or GalX.

59 BASIC BIOLOGICAL SCIENCES↗

Development of genetic tools for heterologous protein expression in a pentose‐utilizing environmental isolate of Pseudomonas putida

Abstract Pseudomonas putida has emerged as a promising host for the conversion of biomass‐derived sugars and aromatic intermediates into commercially relevant biofuels and bioproducts. Most of the strain development studies previously published have focused on P. putida KT2440, which has been engineered to produce a variety of non‐native bioproducts. However, P. putida is not capable of metabolizing pentose sugars, which can constitute up to 25% of biomass hydrolysates. Related P. putida isolates that metabolize a larger fraction of biomass‐derived carbon may be attractive as complementary hosts to P. putida KT2440. Here we describe genetic tool development for P. putida M2, a soil isolate that can metabolize pentose sugars. The functionality of five inducible promoter systems and 12 ribosome binding sites was assessed to regulate gene expression. The utility of these expression systems was confirmed by the production of indigoidine from C6 and C5 sugars. Chromosomal integration and expression of non‐native genes was achieved by using chassis‐independent recombinase‐assisted genome engineering (CRAGE) for single‐step gene integration of biosynthetic pathways directly into the genome of P. putida M2. These genetic tools provide a foundation to develop hosts complementary to P. putida KT2440 and expand the ability of this versatile microbial group to convert biomass to bioproducts.

59 BASIC BIOLOGICAL SCIENCES↗

Galactomannan utilization by Cellvibrio japonicus relies on a single essential α‐galactosidase encoded by the aga27A gene

Plant mannans are a component of lignocellulose that can have diverse compositions in terms of its backbone and side-chain substitutions. Consequently, the degradation of mannan substrates requires a cadre of enzymes for complete reduction to substituent monosaccharides that can include mannose, galactose, and/or glucose. One bacterium that possesses this suite of enzymes is the Gram-negative saprophyte Cellvibrio japonicus, which has 10 predicted mannanases from the Glycoside Hydrolase (GH) families 5, 26, and 27. Here we describe a systems biology approach to identify and characterize the essential mannan-degrading components in this bacterium. The transcriptomic analysis uncovered significant changes in gene expression for most mannanases, as well as many genes that encode carbohydrate active enzymes (CAZymes) when mannan was actively being degraded. A comprehensive mutational analysis characterized 54 CAZyme-encoding genes in the context of mannan utilization. Growth analysis of the mutant strains found that the man26C, aga27A, and man5D genes, which encode a mannobiohydrolase, α-galactosidase, and mannosidase, respectively, were important for the deconstruction of galactomannan, with Aga27A being essential. Our updated model of mannan degradation in C. japonicus proposes that the removal of galactose sidechains from substituted mannans constitutes a crucial step for the complete degradation of this hemicellulose.

59 BASIC BIOLOGICAL SCIENCES↗

Triose phosphate utilization limitation: an unnecessary complexity in terrestrial biosphere model representation of photosynthesis

Triose phosphates are the principal product of photosynthesis. They are used within the chloroplast for starch synthesis, or translocated to the cytosol where they are used to fuel sucrose synthesis. Use of triose phosphate releases inorganic phosphate, and is under strict metabolic control that matches the supply of triose phosphate from the Calvin-Benson cycle to the demand for carbon by sinks (Heldt & Heldt, 2011; McClain & Sharkey, 2019). However, a low rate of triose phosphate utilization (TPU) can deplete the phosphate pool, restrict ATP synthesis and reduce the availability of ATP to power the Calvin-Benson cycle, thereby limiting photosynthesis (Sharkey, 1985).

54 ENVIRONMENTAL SCIENCES↗

Strategies for the production of cell wall‐deconstructing enzymes in lignocellulosic biomass and their utilization for biofuel production

Summary Microbial cell wall‐deconstructing enzymes are widely used in the food, wine, pulp and paper, textile, and detergent industries and will be heavily utilized by cellulosic biorefineries in the production of fuels and chemicals. Due to their ability to use freely available solar energy, genetically engineered bioenergy crops provide an attractive alternative to microbial bioreactors for the production of cell wall‐deconstructing enzymes. This review article summarizes the efforts made within the last decade on the production of cell wall‐deconstructing enzymes in planta for use in the deconstruction of lignocellulosic biomass. A number of strategies have been employed to increase enzyme yields and limit negative impacts on plant growth and development including targeting heterologous enzymes into specific subcellular compartments using signal peptides, using tissue‐specific or inducible promoters to limit the expression of enzymes to certain portions of the plant or certain times, and fusion of amplification sequences upstream of the coding region to enhance expression. We also summarize methods that have been used to access and maintain activity of plant‐generated enzymes when used in conjunction with thermochemical pretreatments for the production of lignocellulosic biofuels.

09 BIOMASS FUELS↗

Utilization of Hydroturbines in Wastewater Treatment Plants

Wastewater treatment plants (WWTPs) are a significant energy consumer, yet there are several opportunities for implementing on-site power generation systems. Within the treatment process, the high flow rate of effluent is produced and discharged to a nearby water body by gravity. Thus, hydroturbines can be utilized to generate power in such an application due to a difference in elevation and high flow rate. This paper presents a case study of introducing a hydroturbine in a WWTP in Wisconsin and evaluating the power output in addition to determining the energy savings. The WWTP considered in this study has an effluent flow rate of 190 MGD (million gallons per day) and elevation difference of 3 m (10 ft) between the final stage of treatment and the discharge point. Based on the parameters above; hubless rim-drive Kaplan type hydroturbine (RDT) is the optimal choice to be used in such an application. The RDT is designed and optimized by using in-house code. A computational fluid dynamics (CFD) software is applied to evaluate the performance of the proposed model, and the system is simulated through homer software to validate the results generated by the CFD. The expected savings is estimated to be 1564 MWh/yr.

Energy & Fuels↗

A Novel Approach for Bounding the Stress Experienced by the Core of Utility-Scale Printed Circuit Heat Exchangers Under Thermohydraulic Loads

Printed circuit heat exchangers (PCHEs) have rapidly gained popularity since being introduced nearly three decades ago, and they are currently widely deployed in the petrochemical and aviation industry. Their compactness, thermohydraulic efficiency, inherent suitability for high temperature/pressure fluids containment, and demonstrated durability are some of the reasons the nuclear industry is seeking to adopt this technology as well. However, the relatively strict nuclear-related regulatory design code, especially when classified as critical to the safety of the reactors, is posing challenges to adopting the technology. From stress analysis point of view, one undesirable feature of PCHEs is their geometrical complexity, which is implied by their multilength-scale features. As a result, a full-scale model of a utility-scale exchanger cannot simply be solved on a computer because meshing such components results in a vast number of degrees-of-freedom. Here, this work seeks to address the challenge of stress analyses to PCHEs by presenting a method to simplify the geometry of PCHE designs. The models proposed by this work can be practically analyzed on a standard computer and provide a path for implementing ASME design rules. The analyses presented herein are divided into five separate investigations. Each is carried out to incrementally simplify the analyzed model by addressing features such as the shapes of the flow passages, the complex distribution of stress in large components, the three-dimensionality of the stress and strain, the thermal stresses caused by thermohydraulic operation observed experimentally and more.

42 ENGINEERING↗

Mechanistic modeling study on process optimization and precursor utilization with atmospheric spatial atomic layer deposition

Spatial atomic layer deposition (SALD) is a promising technology with the aim of combining the advantages of excellent uniformity and conformity of temporal atomic layer deposition (ALD), and an industrial scalable and continuous process. In this manuscript, an experimental and numerical combined model of atmospheric SALD system is presented. To establish the connection between the process parameters and the growth efficiency, a quantitative model on reactant isolation, throughput, and precursor utilization is performed based on the separation gas flow rate, carrier gas flow rate, and precursor mass fraction. The simulation results based on this model show an inverse relation between the precursor usage and the carrier gas flow rate. With the constant carrier gas flow, the relationship of precursor usage and precursor mass fraction follows monotonic function. The precursor concentration, regardless of gas velocity, is the determinant factor of the minimal residual time. The narrow gap between precursor injecting heads and the substrate surface in general SALD system leads to a low Péclet number. In this situation, the gas diffusion act as a leading role in the precursor transport in the small gap rather than the convection. Fluid kinetics from the numerical model is independent of the specific structure, which is instructive for the SALD geometry design as well as its process optimization.

42 ENGINEERING↗

SU-C-BRB-05: Investigation of Conformal Arc Therapy Utilizing Newly Designed Cobalt 60 Machine

Purpose: A new modernized design for cobalt 60 (Co-60) machines is being developed with a ring type gantry. In this study we investigate the beneficial outcome of the new design for conformal arc therapy for various clinical sites. The new modality was evaluated based on isodose distributions and dose volume histograms as compared to 6MV photon beams from conventional linear accelerators. Methods: Computed tomographic images of seven different SBRT patients were selected from our patient database. All of these cases were previously planned on the Eclipse treatment planning system. New plans for these patients were generated with a modified conformal arc technique using both 6MV and Co-60 beams. The conformal arc was created by the delivery of treatment fields conformal to the target cross-section at every 5 or 10 degrees. The field shape was modified or turned off when it initially passed through a critical structure. Monte Carlo codes, MCBEAM and MCPLAN, were used for the machine head simulation and phantom/patient dose calculation, respectively. In the new Co-60 machine design, the source-to-isocenter distance was 60cm and the treatment head included the Co-60 source, primary collimator, jaws and MLC. Results: For all cases investigated, conformal arc plans utilizing Co-60 beams achieved similar conformity (mean conformity index=1.19) comparing to 6MV photon beams. Isodose distributions were tailored similarly around the PTV; both Co-60 and 6MV plans met our clinical acceptance criteria for the target coverage, and the maximum and minimum target doses. The DVH for the Co-60 plans showed slightly lower doses to the critical structures although the differences were small in most cases. Conclusion: There were no significant dosimetric differences between 6MV and Co-60 plans. Our results confirmed that this new Co-60 design could be a cost-effective machine for advanced radiotherapy due to its low cost, low maintenance and high up time.

60 APPLIED LIFE SCIENCES↗

SU-C-BRB-06: Utilizing 3D Scanner and Printer for Dummy Eye-Shield: Artifact-Free CT Images of Tungsten Eye-Shield for Accurate Dose Calculation

Purpose: To evaluate the effect of a tungsten eye-shield on the dose distribution of a patient. Methods: A 3D scanner was used to extract the dimension and shape of a tungsten eye-shield in the STL format. Scanned data was transferred into a 3D printer. A dummy eye shield was then produced using bio-resin (3D systems, VisiJet M3 Proplast). For a patient with mucinous carcinoma, the planning CT was obtained with the dummy eye-shield placed on the patient’s right eye. Field shaping of 6 MeV was performed using a patient-specific cerrobend block on the 15 x 15 cm{sup 2} applicator. The gantry angle was 330° to cover the planning target volume near by the lens. EGS4/BEAMnrc was commissioned from our measurement data from a Varian 21EX. For the CT-based dose calculation using EGS4/DOSXYZnrc, the CT images were converted to a phantom file through the ctcreate program. The phantom file had the same resolution as the planning CT images. By assigning the CT numbers of the dummy eye-shield region to 17000, the real dose distributions below the tungsten eye-shield were calculated in EGS4/DOSXYZnrc. In the TPS, the CT number of the dummy eye-shield region was assigned to the maximum allowable CT number (3000). Results: As compared to the maximum dose, the MC dose on the right lens or below the eye shield area was less than 2%, while the corresponding RTP calculated dose was an unrealistic value of approximately 50%. Conclusion: Utilizing a 3D scanner and a 3D printer, a dummy eye-shield for electron treatment can be easily produced. The artifact-free CT images were successfully incorporated into the CT-based Monte Carlo simulations. The developed method was useful in predicting the realistic dose distributions around the lens blocked with the tungsten shield.

60 APPLIED LIFE SCIENCES↗

SU-C-201-06: Utility of Quantitative 3D SPECT/CT Imaging in Patient Specific Internal Dosimetry of 153-Samarium with GATE Monte Carlo Package

Purpose: Patient-specific 3-dimensional (3D) internal dosimetry in targeted radionuclide therapy is essential for efficient treatment. Two major steps to achieve reliable results are: 1) generating quantitative 3D images of radionuclide distribution and attenuation coefficients and 2) using a reliable method for dose calculation based on activity and attenuation map. In this research, internal dosimetry for 153-Samarium (153-Sm) was done by SPECT-CT images coupled GATE Monte Carlo package for internal dosimetry. Methods: A 50 years old woman with bone metastases from breast cancer was prescribed 153-Sm treatment (Gamma: 103keV and beta: 0.81MeV). A SPECT/CT scan was performed with the Siemens Simbia-T scanner. SPECT and CT images were registered using default registration software. SPECT quantification was achieved by compensating for all image degrading factors including body attenuation, Compton scattering and collimator-detector response (CDR). Triple energy window method was used to estimate and eliminate the scattered photons. Iterative ordered-subsets expectation maximization (OSEM) with correction for attenuation and distance-dependent CDR was used for image reconstruction. Bilinear energy mapping is used to convert Hounsfield units in CT image to attenuation map. Organ borders were defined by the itk-SNAP toolkit segmentation on CT image. GATE was then used for internal dose calculation. The Specific Absorbed Fractions (SAFs) and S-values were reported as MIRD schema. Results: The results showed that the largest SAFs and S-values are in osseous organs as expected. S-value for lung is the highest after spine that can be important in 153-Sm therapy. Conclusion: We presented the utility of SPECT-CT images and Monte Carlo for patient-specific dosimetry as a reliable and accurate method. It has several advantages over template-based methods or simplified dose estimation methods. With advent of high speed computers, Monte Carlo can be used for treatment planning on a day to day basis.

60 APPLIED LIFE SCIENCES↗

SU-E-T-516: Investigation of a Novel Radiochromic Radiation Reporting System Utilizing the Reduction of Ferric Ion

Purpose To introduce and characterize a new “reverse-Fricke” radiation reporting system utilizing the reduction of ferric ions (Fe{sup 3+}) to ferrous ions (Fe{sup 2+}). Methods Two formulations of the radiochromic reporting system, referred to as A and B, were prepared for investigation. Formulation-A consisted of 14 mM 1,10-phenanthroline, 42 mM ethanol, and 57 mM ammonium ferric oxalate in water. Formulation-B consisted of 27 mM 1,10-phenanthroline, 42 mM ethanol, and 28 mM ammonium ferric oxalate in water. Solutions were prepared immediately prior to irradiation with a Cobalt-60 unit with radiation doses of 0, 1, 5, 10, 15, 20, and 25 Gy. The change in optical density over the visible range of 450–650 nm was measured using a spectrophotometer immediately after irradiation. The effective atomic numbers of the formulations were calculated using Mayneord’s formula. Results Ionizing radiation energy absorbed in the solutions causes the reduction of ferric ions (Fe{sup 3+}) into ferrous ions (Fe{sup 2+}), which then forms a 1:3 red colored complex with 1,10-phenanthroline ([(C{sub 1} {sub 2}H{sub 8}N{sup 2}){sub 3}Fe]{sup 2+}) that can be measured spectrophotometrically. The absorbance spectra of the resulting complex displayed a peak maximum at 512 nm with a greater change in absorbance for Formulation-B after receiving comparable radiation doses. The change in absorbance relative to dose exhibited a linear response up to 25 Gy for both Formulation-A (R{sup 2} = 0.98) and Formulation-B (R{sup 2} = 0.97). The novel formulations were also nearly water equivalent (Zeff = 7.42) with effective atomic numbers of 7.65 and 7.52 and mass densities within 0.2% of water. Conclusion Both formulations displayed visible Fe{sup 2+} complex formation with 1,10-phenanthroline after irradiation using a Cobalt-60 source. The higher sensitivity measured for Formulation-B is attributed to the increase in 1,10-phenanthroline concentration and the increase in the 1,10-phenanthroline to ammonium ferric oxalate ratio. Further investigation of this radiation reporting system in a 3D matrix material is encouraged. NSF GRFP Grant Award #LH-102SPS.

60 APPLIED LIFE SCIENCES↗

SU-E-T-787: Utility of the Two Candidate 192-Ir and 169-Yb HDR Sources for Use with a Novel Direction Modulated Brachytherapy Tandem Applicator for Cervical Cancer Treatment

Purpose: A novel tungsten alloy shielded, MRI-compatible, direction modulated brachytherapy (DMBT) concept tandem applicator, which enables unprecedented intensity modulation, was used to evaluate treatment plan quality improvement over a conventional tandem. The utility of the 192-Ir and 169-Yb HDR sources, for use with the DMBT applicator, was evaluated. Methods: The total diameter of the DMBT tandem applicator is 6.0 mm, which consists of 5.4-mm diameter tungsten alloy and 0.3 mm thick plastic sheath. The tandem has 6 symmetric peripheral 1.3-mm diameter grooves for the source to travel. MCNPX v.2.6 was used to simulate the 192-Ir and 169-Yb sources inside the DMBT applicator. First, TG-43 source parameters were evaluated. Second, 3D dose matrix with 1 mm3 resolution were imported into an in-house-coded inverse optimization treatment planning program to obtain optimal plans for 19 clinical cases. All plans were compared with the standard tandem and ring plans. Prescription dose was 15.0 Gy. All plans were normalized to receive the same HRCTV D90. Results: Generally, the DMBT tandem (and ring) plans were better than the conventional tandem and ring plans for 192-Ir and 169-Yb HDR sources. The mean data of D2cc for bladder, rectum, and sigmoid were 11.65±2.30 Gy, 7.47±3.05 Gy, and 9.84±2.48 Gy for Ir-192 DMBT tandem, respectively. These data for Yb-169 were 11.67±2.26 Gy, 7.44±3.02 Gy, and 9.83±2.38 Gy, respectively. The HR-CTV D98 and V100 were 16.37±1.86 Gy and 97.37 ± 1.92 Gy for Ir-192 DMBT, respectively. The corresponding values for Yb-169 were 16.43±1.86 Gy, and 97.51 ± 1.91 Gy. Plans with the 169-Yb source generally produced more favorable results where V100 increased by 13.65% while D2cc across all OARs reduced by 0.54% compared with the 192-Ir plans. Conclusion: For the DMBT tandem applicator, 169-Yb source seems to produce more directional beams resulting in increased intensity modulation capacity, thus resulting in more conformal plans.

60 APPLIED LIFE SCIENCES↗

MO-FG-207-03: Maximizing the Utility of Integrated PET/MRI in Clinical Applications

The use of integrated PET/MRI systems in clinical applications can best benefit from understanding their technological advances and limitations. The currently available clinical PET/MRI systems have their own characteristics. Thorough analyses of existing technical data and evaluation of necessary performance metrics for quality assurances could be conducted to optimize application-specific PET/MRI protocols. This Symposium will focus on technical advances and limitations of clinical PET/MRI systems, and how this exciting imaging modality can be utilized in applications that can benefit from both PET and MRI. Learning Objectives: To understand the technological advances of clinical PET/MRI systems To correctly identify clinical applications that can benefit from PET/MRI To understand ongoing work to further improve the current PET/MRI technology Floris Jansen is a GE Healthcare employee.

60 APPLIED LIFE SCIENCES↗

MO-F-CAMPUS-T-04: Utilization of Optical Dosimeter for Modulated Spot-Scanning Particle Beam

Purpose: To present the utilization of an optical dosimeter for modulated spot-scanning carbon-ion and proton beams during the acceptance test of Siemens IONTRIS system. Method and Materials: An optical dosimeter using phosphor scintillation was developed to map and interactively analyze the shapes and sizes of spots over 190 energies for ProTom modulated-scanning system. The dose response to proton had been characterized with proper pixel calibration at ProTom system. The dose response was further studied at 0.7 cm depths by uniform 8cm in-diameter fields of 424.89 MeV/u (E290) carbon-ions and 215.18MeV (E282) protons at IONTRIS system. The virtual source axial distances (vSAD) of carbonions and protons of IONTRIS system was investigated by measuring either variations of spot position or field size at five different locations to Isocenter. By measuring lateral profiles of uniform doses with varied thin-thicknesses of chest-board pattern and placing the scintillation plate at near to the distal edge, range variations at different off-axis-distances (rOAD) were examined. Relative accuracy and reproducibility of beam range were measured for three beam ranges with a ramping block at front of scintillation plate. Results: Similar dose response was observed for high energies of carbon ions and protons. Mean vSAD at X and Y axes were 744.1 cm and 807.4cm with deviation of 7.4cm and 7.7cm, respectively. Variation of rOAD was within 0.35 mm over 10cm for both protons and carbon ions. Accuracy of measuring relative distal range using the ramping block was 0.2mm. Measured range over repeated three times for each range were within 0.25mm at same room, and within 1.0mm between four rooms. Conclusions: The optical dosimeter could efficiently measure the virtual source distance. And, to measure small range variation at different off-axial locations, and for the relative beam range between rooms during acceptance test of a modulated spot-scanning particle system.

60 APPLIED LIFE SCIENCES↗