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At least 37 records · Page 2

Hard X-ray self-seeding commissioning at PAL-XFEL

A wake monochromator based on a large-area diamond single crystal for hard X-ray self-seeding has been successfully installed and commissioned in the hard X-ray free-electron laser (FEL) at the Pohang Accelerator Laboratory with international collaboration. For this commissioning, the self-seeding was demonstrated with a low bunch charge (40 pC) and the nominal bunch charge (180 pC) of self-amplified spontaneous emission (SASE) operation. The FEL pulse lengths were estimated as 7 fs and 29.5 fs, respectively. In both cases, the average spectral brightness increased by more than three times compared with the SASE mode. The self-seeding experiment was demonstrated for the first time using a crystal with a thickness of 30 µm, and a narrow bandwidth of 0.22 eV (full width at half-maximum) was obtained at 8.3 keV, which confirmed the functionality of a crystal with such a small thickness. In the nominal bunch-charge self-seeding experiment, the histogram of the intensity integrated over a 1 eV bandwidth showed a well defined Gaussian profile, which is evidence of the saturated FEL and a minimal electron-energy jitter (~1.2 × 10 –4 ) effect. Here, the corresponding low photon-energy jitter (~2.4 × 10 –4 ) of the SASE FEL pulse, which is two times lower than the Pierce parameter, enabled the seeding power to be maximized by maintaining the spectral overlap between SASE FEL gain and the monochromator.

47 OTHER INSTRUMENTATION↗

Brachypodium Phenylalanine Ammonia Lyase (PAL) Promotes Antiviral Defenses against Panicum mosaic virus and Its Satellites

Brachypodium distachyon has recently emerged as a premier model plant for monocot biology, akin to Arabidopsis thaliana. We previously reported genome-wide transcriptomic and alternative splicing changes occurring in Brachypodium during compatible infections with Panicum mosaic virus (PMV) and its satellite virus (SPMV). Here, we dissected the role of Brachypodium phenylalanine ammonia lyase 1 (PAL1), a key enzyme for phenylpropanoid and salicylic acid (SA) biosynthesis and the induction of plant defenses. Targeted metabolomics profiling of PMV-infected and PMV- plus SPMV-infected (PMV/SPMV) Brachypodium plants revealed enhanced levels of multiple defense-related hormones and metabolites such as cinnamic acid, SA, and fatty acids and lignin precursors during disease progression. The virus-induced accumulation of SA and lignin was significantly suppressed upon knockdown of B. distachyon PAL1 (BdPAL1) using RNA interference (RNAi). The compromised SA accumulation in PMV/SPMV-infected BdPAL1 RNAi plants correlated with weaker induction of multiple SA-related defense gene markers (pathogenesis related 1 [PR-1], PR-3, PR-5, and WRKY75) and enhanced susceptibility to PMV/SPMV compared to that of wild-type (WT) plants. Furthermore, exogenous application of SA alleviated the PMV/SPMV necrotic disease phenotypes and delayed plant death caused by single and mixed infections. Together, our results support an antiviral role for BdPAL1 during compatible host-virus interaction, perhaps as a last resort attempt to rescue the infected plant.

59 BASIC BIOLOGICAL SCIENCES↗

PAL Experimental Report Form

We constructed the first simultaneous dark- and bright-field X-ray microscope in this experiment, coupling it to simultaneous measurements of the wide-angle Xray diffraction, and the XFEL pulse intensity and energy spectrum on each shot. This experiment established proof of concept for this new advanced measurement technique and demonstrated DFXM at an XFEL for the first time. Within this complex experiment, we also acquired data that advanced our research goals within materials science and mathematics. These included: A preliminary study of how local effects reduce the X-ray damage threshold in diamond; A preliminary study of how X-ray radiation imparts heat into materials based on irreversible phase transitions that occur in boric acid with increasing temperature (based on hydration); A study of X-ray radiation damage from ultrafast thermal cycling and nonthermal melting in bismuth selenide; We also began developing software to automate alignment of compound refractive lenses (CRLs) using stochastic Nelder-Mead optimization methods. This work will expedite and improve the accuracy of alignments for many future experiments using CRLs.

36 MATERIALS SCIENCE↗

Metabolic targeting with recombinant methioninase combined with palbociclib regresses a doxorubicin-resistant dedifferentiated liposarcoma

Highlights: • Ddedifferentiated liposarcoma (DDLPS) is recalcitrant and has the lowest survival rate. • Determine the efficacy of rMETase combined with palbociclib (PAL) against a DOX-resistant DDLPS in a PDOX mouse model. • The combination of PAL and rMETase significantly regressed tumor volume. • The combination of rMETase and PAL could be developed clinically. Liposarcoma is the most common type of soft tissue sarcoma. Among the subtypes of liposarcoma, dedifferentiated liposarcoma (DDLPS) is recalcitrant and has the lowest survival rate. The aim of the present study is to determine the efficacy of metabolic targeting with recombinant methioninase (rMETase) combined with palbociclib (PAL) against a doxorubicin (DOX)-resistant DDLPS in a patient-derived orthotopic xenograft (PDOX) model. A resected tumor from a patient with recurrent high-grade DDLPS in the right retroperitoneum was grown orthotopically in the right retroperitoneum of nude mice to establish a PDOX model. The PDOX models were randomized into the following groups when tumor volume reached 100 mm{sup 3}: G1, control without treatment; G2, DOX; G3, PAL; G4, recombinant methioninase (rMETase); G5, PAL combined with rMETase. Tumor length and width were measured both pre- and post-treatment. On day 14 after initiation, all treatments significantly inhibited tumor growth compared to the untreated control except DOX. PAL combined with rMETase was significantly more effective than both DOX, rMETase alone, and PAL alone. Combining PAL and rMETase significantly regressed tumor volume on day 14 after initiation of treatment and was the only treatment to do so. The relative body weight on day 14 compared with day 0 did not significantly differ between each treatment group. The results of the present study indicate the powerful combination of rMETase and PAL should be tested clinically against DDLPS in the near future.

60 APPLIED LIFE SCIENCES↗

THE PROPER MOTION OF PALOMAR 5

Palomar 5 (Pal 5) is a faint halo globular cluster associated with narrow tidal tails. It is a useful system to understand the process of tidal dissolution, as well as to constrain the potential of the Milky Way. A well-determined orbit for Pal 5 would enable detailed study of these open questions. We present here the first CCD-based proper motion measurement of Pal 5 obtained using SDSS as a first epoch and new Large Binocular Telescope/Large Binocular Camera (LBC) images as a second, giving a baseline of 15 years. We perform relative astrometry, using SDSS as a distortion-free reference, and images of the cluster and also of the Pal 5 stream for the derivation of the distortion correction for LBC. The reference frame is made up of background galaxies. We correct for differential chromatic refraction using relations obtained from SDSS colors as well as from flux-calibrated spectra, finding that the correction relations for stars and for galaxies are different. We obtain μ{sub α} = −2.296 ± 0.186 mas yr{sup −1} and μ{sub δ} = −2.257 ± 0.181 mas yr{sup −1} for the proper motion of Pal 5. We use this motion, and the publicly available code galpy, to model the disruption of Pal 5 in different Milky Way models consisting of a bulge, a disk, and a spherical dark matter halo. Our fits to the observed stream properties (streak and radial velocity gradient) result in a preference for a relatively large Pal 5 distance of around 24 kpc. A slightly larger absolute proper motion than what we measure also results in better matches but the best solutions need a change in distance. We find that a spherical Milky Way model, with V{sub 0} = 220 km s{sup −1} and V{sub 20} {sub kpc}, i.e., approximately at the apocenter of Pal 5, of 218 km s{sup −1}, can match the data well, at least for our choice of disk and bulge parametrization.

79 ASTRONOMY AND ASTROPHYSICS↗

Arabidopsis SnRK1 negatively regulates phenylpropanoid metabolism via Kelch domain-containing F-box proteins

Phenylpropanoid metabolism represents a substantial metabolic sink for the photosynthetically fixed carbon. The evolutionarily conserved Sucrose Non-Fermenting Related Kinase 1 (SnRK1) is a major metabolic sensor that reprograms metabolism upon carbon deprivation. However, it is not clear if and how SnRK1-mediated sugar signaling pathway controls phenylpropanoid metabolism. Here, we show that Arabidopsis SnRK1 negatively regulates phenylpropanoid biosynthesis via a group of Kelch domain-containing F-Box (KFB) proteins that are responsible for the ubiquitination and degradation of phenylalanine ammonia lyase (PAL). Down-regulation of AtSnRK1 significantly promoted the accumulation of soluble phenolics and lignin polymers and drastically increased PAL cellular accumulation but only slightly altered its transcription level. Co-expression of SnRK1α with PAL in Nicotiana benthamiana leaves resulted in the severe attenuation of the latter's protein level, but protein interaction assays suggested PAL is not a direct substrate of SnRK1. Furthermore, up- or down-regulation of AtSnRK1 positively affected KFB PALs gene expression, and energy starvation upregulated KFB PAL expression, which partially depends on AtSnRK1. Collectively, our study reveals that SnRK1 negatively regulates phenylpropanoid biosynthesis, and KFB PALs act as regulatory components of the SnRK1 signaling network, transcriptionally regulated by SnRK1 and subsequently mediating proteasomal degradation of PAL in response to the cellular carbon availability.

59 BASIC BIOLOGICAL SCIENCES↗

Zeolite‐Based Catalysts for Conversion of Oxygenated Polymer Waste by Positron Annihilation

Positron Annihilation Lifetime Spectroscopy (PALS) has been employed to investigate the catalysts HZSM-5 and MESO−Y, which play a pivotal role in catalyzing and upgrading plastics, with a primary focus on oxygenated polymers, thereby transforming existing plastic materials into simpler, higher-quality value-added products. Here, in this study, PALS was systematically compared with other complementary analytical techniques. The research outcomes have successfully demonstrated the efficacy of PALS in elucidating the morphology and topology of zeolites at micro/meso-meter scales. The first experiment focuses on H-ZSM-5 zeolite subjected to treatments involving polyurethane and polypropylene. The second experiment delves into H-ZSM-5 zeolites with varying Si/Al ratios, both before and after conversion. The third experiment investigates Y zeolites that are surfactant templated to induce meso-porosity, examining their fresh state as well as their post-conversion condition. The PALS analysis was supplemented by BET (Brunauer-Emmett-Teller) analysis and NMR (Nuclear Magnetic Resonance) spectroscopies. Notably, PALS exhibits superior sensitivity, at the sub-nanometer scale, suggesting its potential as a preferred complementary method for catalysis studies. In conclusion, the integration of PALS into the repertoire of analytical tools enhances our understanding of catalyst behavior and catalytic processes, offering valuable insights for the advancement of plastic recycling and catalysis research.

32 - ENERGY CONSERVATION, CONSUMPTION, AND UTILIZA↗

Pancreatic stellate cell activation is regulated by fatty acids and ER stress

Introduction: Pancreatic pathologies are characterized by a progressive fibrosis process. Pancreatic stellate cells (PSC) play a crucial role in pancreatic fibrogenesis. Endoplasmic reticulum (ER) stress emerges as an important determinant of fibrotic remodeling. Overload of fatty acids (FA), typical to obesity, may lead to lipotoxic state and cellular stress. Aim: To study the effect of different lipolytic challenges on pancreatic ER stress and PSC activation. Methods: Primary PSCs were exposed to different FAs, palmitate (pal) and oleate (ole), at pathophysiological concentrations typical to obese state, and in acute caerulein-induced stress (cer). PSC activation and differentiation were analyzed by measuring fat accumulation (oil-red staining and quantitation), proliferation (cells count) and migration (wound- healing assay). PSC differentiation markers (α-sma, fibronectin, tgf-β and collagen secretion), ER stress unfolded protein response and immune indicators (Xbp1, CHOP, TNF-α, IL-6) were analyzed at the transcript and protein expression levels (quantitative RT-PCR and western blotting). Results: PSC exposure to pal and ole FAs (500 µM) increased significantly fat accumulation. Proliferation and migration analysis demonstrated that ole FA retained PSC activation, while exposure to pal FA significantly halted proliferation rate and delayed migration. Cer significantly augmented PSC differentiation markers α- sma, fibronectin and collagen, and ER stress and inflammation markers including Xbp1, CHOP, TNF-α and IL-6. The ole FA treatment significantly elevated PSC differentiation markers α-sma, fibronectin and collagen secretion. PSC ER stress was demonstrated following pal treatment with significant elevation of Xbp1 splicing and CHOP levels. Conclusion: Exposure to pal FA halted PSC activation and differentiation and elevated ER stress markers, while cer and ole exposure significantly induced activation, differentiation and fibrosis. Thus, dietary FA composition should be considered and optimized to regulate PSC activation and differentiation in pancreatic pathologies. - Highlights: • Pancreatic stellate cells (PSC) play a crucial role in pancreatic fibrogenesis. • Endoplasmic reticulum (ER) stress is an important determinant of fibrotic remodeling. • Overload of fatty acids, typical to obesity, may lead to lipotoxicity and cellular stress. • Exposure to pal FA halts PSC activation, differentiation and elevated ER stress. • Cer and ole exposure significantly induced activation, differentiation and fibrosis.

60 APPLIED LIFE SCIENCES↗

Complete biosynthesis of salicylic acid from phenylalanine in plants

Salicylic acid (SA) is a pivotal phytohormone for plant responses to biotic and abiotic stresses. Plants have evolved two pathways to produce SA: the isochorismate synthase and phenylalanine ammonia lyase (PAL) pathways. Whereas the isochorismate synthase pathway has been fully identified, the PAL pathway remains incomplete. Here we report the full characterization of the PAL pathway for SA biosynthesis via functional analysis of rice (Oryza sativa) SA-DEFICIENT GENE 1 (OSD1) to OSD4. The cinnamoyl-coenzyme A (CoA) ligase OSD1 catalyses the conversion of trans-cinnamic acid to cinnamoyl-CoA, which is subsequently transformed to benzoyl-CoA via the β-oxidative pathway in peroxisomes. The resulting benzoyl-CoA is further converted to benzyl benzoate by the peroxisomal benzoyltransferase OSD2. Benzyl benzoate is subsequently hydroxylated to benzyl salicylate by the endoplasmic reticulum membrane-resident cytochrome P450 OSD3, which is ultimately hydrolysed to salicylic acid by the cytoplasmic carboxylesterase OSD4. Evolutionary analyses reveal that the PAL pathway was first assembled before the divergence of gymnosperms and has been conserved in most seed plants. Activation of the PAL pathway in rice significantly enhances salicylic acid levels and plant immunity. Completion of the PAL pathway provides critical insights into the primary salicylic acid biosynthetic pathway across plant species and offers a precise target for modulating crop immunity.

59 BASIC BIOLOGICAL SCIENCES↗

A 6 MW/m 2 High Heat Flux Testing Facility of Irradiated Materials Using Infrared Plasma-Arc Lamps

Assessing the effect of neutron irradiation of plasma-facing materials has been challenging due to both the technical and radiological challenges involved. In an effort to address the radiological challenges, a facility was developed to conduct high heat flux testing (HHFT) of inherently small samples of neutron-irradiated materials. A new line-focus reflector was designed and fabricated at Oak Ridge National Laboratory for a plasma-arc lamp (PAL) to attain a source heat flux of 12 MW/m 2 . The new reflector was fabricated with two ports for monitoring specimen condition during HHFT. At the same operational conditions for PAL, the absorbed heat flux in tungsten was increased from 1.39 MW/m 2 with the uniform irradiance reflector to 5.12 MW/m 2 for the line-focus reflector. This fourfold increase in the heat flux, at the same PAL electrode lifetimes, enabled cost-effective facility operation for a high number of cyclic high heat flux tests. Specifically, the test section is confined to a hemispherical dome, and specimens are bolted directly to a water-cooled copper alloy rod. Temperature measurement in the PAL facility was a main challenge due to a limited line of sight. For the first time in a PAL facility operating at high heat fluxes, the specimen surface temperature was directly measured during HHFT with a pyrometer. Finally, the HHFT data, which were obtained in this upgraded PAL facility, demonstrated the facility readiness for irradiated materials.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

DETAILED CHEMICAL ABUNDANCES OF FOUR STARS IN THE UNUSUAL GLOBULAR CLUSTER PALOMAR 1

Detailed chemical abundances for 21 elements are presented for four red giants in the anomalous outer halo globular cluster Palomar 1 (R{sub GC} = 17.2 kpc, Z = 3.6 kpc) using high-resolution (R = 36, 000) spectra from the High Dispersion Spectrograph on the Subaru Telescope. Pal 1 has long been considered unusual because of its low surface brightness, sparse red giant branch, young age, and its possible association with two extragalactic streams of stars. This paper shows that its chemistry further confirms its unusual nature. The mean metallicity of the four stars, [Fe/H] = -0.60 {+-} 0.01, is high for a globular cluster so far from the Galactic center, but is low for a typical open cluster. The [{alpha}/Fe] ratios, though in agreement with the Galactic stars within the 1{sigma} errors, agree best with the lower values in dwarf galaxies. No signs of the Na/O anticorrelation are detected in Pal 1, though Na appears to be marginally high in all four stars. Pal 1's neutron-capture elements are also unusual: its high [Ba/Y] ratio agrees best with dwarf galaxies, implying an excess of second-peak over first-peak s-process elements, while its [Eu/{alpha}] and [Ba/Eu] ratios show that Pal 1's contributions from the r-process must have differed in some way from normal Galactic stars. Therefore, Pal 1 is unusual chemically, as well in its other properties. Pal 1 shares some of its unusual abundance characteristics with the young clusters associated with the Sagittarius dwarf galaxy remnant and the intermediate-age LMC clusters, and could be chemically associated with the Canis Majoris overdensity; however, it does not seem to be similar to the Monoceros/Galactic Anticenter Stellar Stream.

79 ASTRONOMY AND ASTROPHYSICS↗

Down-regulation of kelch domain-containing F-box protein in Arabidopsis enhances the production of (poly)phenols and tolerance to ultraviolet radiation

Phenylpropanoid biosynthesis in plants engenders myriad phenolics with diverse biological functions. Phenylalanine ammonia-lyase (PAL) is the first committed enzyme in the pathway, directing primary metabolic flux into a phenylpropanoid branch. Previously, we demonstrated that the Arabidopsis Kelch-domain containing F-box proteins, AtKFB01, -20, and -50, function as the negative regulators controlling phenylpropanoid biosynthesis via mediating PAL’s ubiquitination and subsequent degradation. Here, we reveal that Arabidopsis KFB39, a close homolog of AtKFB50, also interacts physically with PAL isozymes and modulates PALs' stability and activity. Disturbing the expression of KFB39 reciprocally affects the accumulation/deposition of a set of phenylpropanoid end products, suggesting that KFB39 is an additional post-translational regulator responsible for the turnover of PAL and negatively controlling phenylpropanoid biosynthesis. Furthermore, we discover that exposure of Arabidopsis to UV-B radiation suppresses the expression of all four KFB genes while inducing the transcription of PAL isogenes; these data suggest that Arabidopsis consolidates both transcriptional and post-translational regulation mechanisms to maximize its responses to UV stress. Simultaneous down-regulation of all four identified KFBs significantly enhances the production of (poly)phenols and the plant’s tolerance to UV irradiation. This study offers a biotechnological approach for engineering the production of useful phenolic chemicals and for increasing a plant’s resistance to environmental stress.

59 BASIC BIOLOGICAL SCIENCES↗