Radiation Damage Research Articles

This study investigates the effects of ultrasmall (∼4nm)gold nanoparticles (AuNPs) combined with X-ray irradiation to enhanceradiotherapy efficacy. Using the in vivo Drosophilamelanogaster model, we observed that while AuNPs alonedelayed embryonic development, their combination with irradiationcompletely halted it. Lifespan analysis showed that irradiated fliesfed with AuNPs had a slight survival advantage, suggesting a protectiveeffect against radiation-induced oxidative stress. Immunofluorescenceanalysis revealed increased DNA damage (or repair) in the flies, supportingthe potential of AuNPs to boost the local radiation dose and offerprotection against radiation-induced damage, with implications foroptimized therapeutic strategies.

Abstract: Radiation therapy is a cornerstone of cancer treatment, yet its efficacy is often compromised by severe side effects that damage healthy tissues, leading to significant patient discomfort and treatment interruptions. Inspired by the extraordinary resilience of tardigrades, microscopic organisms capable of surviving extreme conditions, researchers have developed a novel approach to mitigate radiation-induced damage. This study focuses on the tardigrade-derived protein Dsup (Damage suppressor), which protects DNA from radiation. By delivering messenger RNA (mRNA) encoding the damage suppressor (Dsup), a DNA-binding protein found in tardigrades, via specialized nanoparticles, scientists have successfully reduced radiation-induced DNA damage in mouse models by up to 50%. The localized and temporary expression of Dsup ensures that healthy tissues are protected without compromising the effectiveness of radiation on tumors. This innovative strategy not only enhances the safety and tolerability of radiation therapy but also holds promise for broader applications, potentially relevant in other high-radiation exposure scenarios, such as chemotherapy or space missions. However, these applications remain to be thoroughly investigated. The research conducted by a collaborative team from MIT, Brigham and Women';s Hospital, and the University of Iowa represents a significant advancement in cancer treatment, offering a potential paradigm shift in how we approach radiation damage mitigation. Future efforts will focus on optimizing the delivery system and adapting the Dsup protein for human use, paving the way for clinical trials and real-world applications. This breakthrough underscores the potential of bio-inspired solutions in addressing complex medical challenges.

The high levels of flux at a fourth-generation synchrotron are shown to have significant beam heating effects with increasing risk of radiation damage X-ray crystallography technique is widely used to determine the three-dimensional structures of macromolecules and it is very important to know and how it might affect an X- ray diffraction experiments and the resulting structures. There are two types of radiation damages (global and specific).The radiation damage process is dose dependent and there is no technique available to prevent. X- ray crystallography can be used to monitor the damages by collecting consecutive data sets in the same protein crystal to track the progression of damages. Damage incurred during data collection in macromolecular crystallography (MX) limits the information that can be obtained from a single crystal, and it may also prevent getting the solution of structure. Each year, hundreds of scientists are using SER-CAT to conduct X-ray diffraction experiments many of which are directly related to human diseases. Their efficiency and quality of the data collected for various high impact scientific projects will be depending upon the how precisely we study this radiation damage and provide a general data collection strategy especially for the increased intensity (after APS-U) to SER-CAT user community. To study the consequences and progression of radiation damages, 10 consecutive datasets were collected using single trypsin crystal and analyzed. The results for both global and specific damage effects will be presented.

RAPD2 is a modular package of programs written for the automated processing of macromolecular crystallographic data at the NE-CAT beamlines. It monitors for collected data, processes snapshots to create strategies for data collection, processes data runs for structure solution, and can then solve the structure using molecular replacement or single-wavelength anomalous diffraction with results stored in a MongoDB. Most of the backend code is written in Python3 with an AngularJS based frontend. This allows users to login with a web browser to view results, modify settings and rerun jobs, or launch additional pipelines (see Kay Perry). The RAPD2 code is designed to be modular on multiple levels. With a variety of possible experimental and computing environments in mind, RAPD2 is separated into several interdependent modules. At the highest level, X-Ray source monitoring (Monitors), core data handling and archiving (Control), and job launching (Launch) can be started through the Control or separated into distinct programs that communicate by passing Python objects over standard TCP sockets or Redis Streams. This allows flexibility in setup; for example, RAPD was developed on a single computer, so the Control and the Launch programs initially ran on a single machine; when a computational cluster was later acquired, the Launch module was moved with minimal changes to either program. On a deeper level, the code is divided into functionally distinct modules. For example, each pipeline is self-contained and called by a launch adapter when needed, allowing nimble development (i.e. bug fixes) that do not require any program restarting to take effect. Additional benefits of the built-in modularity are that all site-specific settings and functions are isolated as much as possible to one module (Site), so adapting RAPD to a new experimental environment is relatively simple. At NE-CAT, Redis Streams are used for communication between the beamline and the RAPD2 Monitors; however this can be modified to suit different environments at other facilities. The Monitors save the beamline information in MongoDB and pass it to Control, where the strategy or data processing commands are generated. These commands are sent to a Launch manager that decides where to launch the job based on Site settings. This provides flexibility in launching jobs on specific machines using a shell launch adapter or on a computer cluster with launch adapters for various cluster workload managers including SLURM, SGE, and PBS. For strategy commands, the index pipeline is launched, where six Labelit autoindexing jobs are started simultaneously with different peak pick settings, each optimized for different types of diffraction data. Once the best solution is determined, Raddose is run to calculate radiation damage parameters, which are input to BEST for the regular and anomalous data collection strategies. This pipeline takes approximately 30s to complete, and results are displayed in the UI. The index pipeline is modular, so other auto-index, radiation damage, or strategy programs can be launched through different plugins. For data runs, the integration pipeline is launched running XDS multiple times to optimize the results. This pipeline takes a few minutes to finish, and results are displayed in the UI. A new mintegrate pipeline will additionally launch data processing in autoPROC, Fast DP, and XIA2, with results from all 4 data processing programs displayed in the UI for comparison.

In the high-luminosity era of the Large Hadron Collider, the instantaneous luminosity is expected to reach unprecedented values, resulting in up to 200 proton-proton interactions in a typical bunch crossing. To cope with the resulting increase in occupancy, bandwidth and radiation damage, the ATLAS Inner Detector will be replaced by an all-silicon system, the Inner Tracker (ITk). The innermost part of the ITk will consist of a pixel detector, with an active area of about 13 m2. To deal with the changing requirements in terms of radiation hardness, power dissipation and production yield, several silicon sensor technologies will be employed in the five barrel and endcap layers. As a timeline, it is facing to pre-production of components, sensor, building modules, mechanical structures and services. The pixel modules assembled with RD53B readout chips have been built to evaluate their production rate. Irradiation campaigns were done to evaluate their thermal and electrical performance before and after irradiation. A new powering scheme — serial — will be employed in the ITk pixel detector, helping to reduce the material budget of the detector as well as power dissipation. This contribution presents the status of the ITk-pixel project focusing on the lessons learned and the biggest challenges towards production, from mechanics structures to sensors, and it will summarize the latest results on closest-to-real demonstrators built using module, electric and cooling services prototypes.

Human manganese superoxide dismutase (MnSOD2) is a metallo-oxidoreductase localized to the mitochondrial matrix. Its canonical function is as an antioxidant, neutralizing superoxide radicals generated during the electron transport chain (ETC). By converting superoxide into hydrogen peroxide and molecular oxygen, MnSOD2 safeguards sensitive metabolic enzymes from oxidative damage and facilitates mitochondrial redox signaling via membrane-diffusible hydrogen peroxide. At the core of MnSOD2’s activity is a proton-coupled electron transfer (PCET) mechanism driven by the cyclical oxidation and reduction of the active-site Mn.The clinical relevance of MnSOD2 is contradictory: downregulation promotes tumorigenesis, while upregulation promotes increased malignancy and metastatic activity in established tumors. Recent investigations into the mechanism underpinning this dichotomy have suggested the erroneous metal incorporation could be to blame. When MnSOD2 is overexpressed, enzyme levels outstrip intracellular Mn reserves and it begins incorporating Fe in lieu of Mn, becoming FeSOD2. While FeSOD2 has long been known to be catalytically dead towards superoxide, recent studies have uncovered it gains prooxidant peroxidase activity instead. Clinically, FeSOD2 has been observed acting as a stemness- promoting histone demethylase in breast cancer.Despite its importance, little headway has been made towards understanding the atomic mechanism of this catalytic shift from SOD to peroxidase. One of the major challenges in elucidating the mechanism is the inherent difficulty of studying metallo-oxidoreductases and PCETs using traditional structural techniques like X-ray crystallography. X-rays often reduce sensitive metal centers, preventing the uncovering of the mechanically critical oxidized intermediates. Furthermore, protons are insensitive to X-ray diffraction, making the technique unsuited for identifying labile protons in PCET reactions. We aim to overcome these issues using neutron crystallography. Neutrons are non-ionizing, making them safe to use on oxidized metal centers as they won’t cause radiation damage or photoreduction. Hydrogens also have a similar scattering cross-section to carbon, making them readily apparent in nuclear density maps. This ability to directly image proton locations makes neutron diffraction ideal for studying PCET mechanisms.Our lab has previously used macromolecular neutron diffraction studies to elucidate the PCETs underpinning the mechanism of MnSOD2, and we now seek to use a similar methodology to uncover how the atomic mechanism changes upon Fe incorporation. In preparation for our neutron studies, we have solved X-ray diffraction structures of oxidized, reduced, and H¬¬2O2-bound FeSOD2 to provide a foundation for interpreting future neutron crystallographic data. Additionally, we used X-ray absorption spectroscopy (XAS) to determine the molecular orbitals, coordination state, oxidation number, and interatomic distances of the metal center of oxidized, reduced, and substrate-bound FeSOD2. Along with directly informing the electronic state of the metal in catalytically relevant states, this data will provide refinement constraints for our neutron diffraction structures.Our X-ray crystallography data shows that we can successfully manipulate the redox state of FeSOD2 crystals, and that the substrate binds to the Fe center. XAS data shows that the metal center can oscillate between 2+ and 3+ oxidation states. These results are encouraging for our future neutron crystallography and indicate a high likelihood of success. By elucidating this mechanism, we hope to improve our understanding of redox biology and present a potential therapeutic target for SOD2-enriched malignancies.

Hydrogen sulfide (H2S) has emerged as an important biological signaling molecule. Its interaction with insulin impacts on glucose and lipid metabolisms. However, the molecular mechanisms underlying the cellular effects of H2S have been unsettled. To obtain direct evidence for H2S-induced post-translational modification of insulin molecule, we structurally characterized insulin following incubation with NaHS, an H2S salt, using X-ray crystallography. Insulin crystals were grown using the hanging drop vapor diffusion method and optimized in MES buffer with PEG MME550 and zinc sulfate. X-ray diffraction data were collected at the Canadian Light Source to resolutions between 2.1–2.2 Å. Six datasets were processed, structures solved and refined (representative structure PDB 9MRA; Rwork/Rfree 0.18/0.23). Structural analysis showed electron density corresponding to a sulfur atom near the amide group of Gln4 in chain B, aligning with prior LC-MS predictions. However, N–S distances ranged from 2.35–3.4 Å across the structures, suggesting the possibility of a transient covalent interaction disrupted by radiation damage during data collection. In addition, one structure exhibited a sulfur atom near Glu residues, suggesting secondary or alternative binding sites. All refined structures were predicted to form hexameric assemblies based on PISA analysis. These results provide structural support to H2S-induced post-translational modification of the insulin molecule.

Human manganese superoxide dismutase (MnSOD2) is a metallo-oxidoreductase localized to the mitochondrial matrix. Its canonical function is as an antioxidant, neutralizing superoxide radicals generated during the electron transport chain (ETC). By converting superoxide into hydrogen peroxide and molecular oxygen, MnSOD2 safeguards sensitive metabolic enzymes from oxidative damage and facilitates mitochondrial redox signaling via membrane-diffusible hydrogen peroxide. At the core of MnSOD2’s activity is a proton-coupled electron transfer (PCET) mechanism driven by the cyclical oxidation and reduction of the active-site Mn.The clinical relevance of MnSOD2 is contradictory: downregulation promotes tumorigenesis, while upregulation promotes increased malignancy and metastatic activity in established tumors. Recent investigations into the mechanism underpinning this dichotomy have suggested the erroneous metal incorporation could be to blame. When MnSOD2 is overexpressed, enzyme levels outstrip intracellular Mn reserves and it begins incorporating Fe in lieu of Mn, becoming FeSOD2. While FeSOD2 has long been known to be catalytically dead towards superoxide, recent studies have uncovered it gains prooxidant peroxidase activity instead. Clinically, FeSOD2 has been observed acting as a stemness- promoting histone demethylase in breast cancer.Despite its importance, little headway has been made towards understanding the atomic mechanism of this catalytic shift from SOD to peroxidase. One of the major challenges in elucidating the mechanism is the inherent difficulty of studying metallo-oxidoreductases and PCETs using traditional structural techniques like X-ray crystallography. X-rays often reduce sensitive metal centers, preventing the uncovering of the mechanically critical oxidized intermediates. Furthermore, protons are insensitive to X-ray diffraction, making the technique unsuited for identifying labile protons in PCET reactions. We aim to overcome these issues using neutron crystallography. Neutrons are non-ionizing, making them safe to use on oxidized metal centers as they won’t cause radiation damage or photoreduction. Hydrogens also have a similar scattering cross-section to carbon, making them readily apparent in nuclear density maps. This ability to directly image proton locations makes neutron diffraction ideal for studying PCET mechanisms.Our lab has previously used macromolecular neutron diffraction studies to elucidate the PCETs underpinning the mechanism of MnSOD2, and we now seek to use a similar methodology to uncover how the atomic mechanism changes upon Fe incorporation. In preparation for our neutron studies, we have solved X-ray diffraction structures of oxidized, reduced, and H¬¬2O2-bound FeSOD2 to provide a foundation for interpreting future neutron crystallographic data. Additionally, we used X-ray absorption spectroscopy (XAS) to determine the molecular orbitals, coordination state, oxidation number, and interatomic distances of the metal center of oxidized, reduced, and substrate-bound FeSOD2. Along with directly informing the electronic state of the metal in catalytically relevant states, this data will provide refinement constraints for our neutron diffraction structures.Our X-ray crystallography data shows that we can successfully manipulate the redox state of FeSOD2 crystals, and that the substrate binds to the Fe center. XAS data shows that the metal center can oscillate between 2+ and 3+ oxidation states. These results are encouraging for our future neutron crystallography and indicate a high likelihood of success. By elucidating this mechanism, we hope to improve our understanding of redox biology and present a potential therapeutic target for SOD2-enriched malignancies.

A program for serial handling of crystallographic data is presented within Olex2. Especially for small molecule electron and x-ray diffraction, the handling of several datasets of the same structure can be tedious and prone to errors, which can affect comparability. The program SISYPHOS allows for the individual refinement of a starting model against several recorded datasets (in “.hkl” format) with adaptation to changes in the unit cell, wavelength, among other parameters. The program was tested for resonant diffraction (also known as anomalous dispersion), investigations on radiation damage, the benchmarking of different configurations for quantum crystallographic modeling, electron diffraction data, and for testing several datasets from the same measurement using various settings to identify the most suitable dataset.

ObjectiveThe feasibility of placing longer, larger diameter double-threaded screws into the pedicle for good fixation in osteoporotic patients with lumbar spondylolisthesis was investigated via robot-assisted optimal access planning.MethodA total of 80 patients with degenerative lumbar spondylolisthesis needed posterior incision decompression and bone grafting combined with pedicle screw fixation due to spondylolisthesis. The patients were equally and randomly assigned to a robot-assisted group and a bone cement-strengthened group. The operative time, intraoperative blood loss, and intraoperative radiation dose were recorded. X-ray and CT scans were routinely reviewed after the surgery. The ratio of screw diameter to pedicle width (SD/PW) was calculated. The pedicle position was graded. The Bub score assessed proximal facet joint invasion. Visual analogue pain scale (VAS) was recorded before surgery and 3days after surgery. The Oswestry Disability Index (ODI) and health Survey Summary Form (SF-36 to assess patients' quality of life) were performed before surgery and 6months after surgery. The rate of screw loosening, removal, complications and revision were evaluated by X-ray and CT 12months after operation.ResultsVAS score on day 3 after surgery was significantly better in the robot-assisted group than in the bone cement-strengthened group. (p = 0.027). The operative time and intraoperative radiation dose of the robot-assisted group were lower than those of the bone cement-strengthened group (p < 0.001). The ratios of screw length, screw diameter, and SD/PW in both groups were significantly better in the robot-assisted group than in the bone cement-strengthened group (p < 0.001). The incidence of screw small joint invasion was 10.2% in the robot-assisted group and 19.1% in the bone cement-strengthened group, with a statistically significant difference between the two (p = 0.020). The Oswestry Disability Index (ODI) and Health Survey Summary Form (SF-36) at 6months after surgery were significantly improved in both groups.ConclusionPatients with osteoporotic lumbar spondylolisthesis who use robot assistance to implant longer, thicker-diameter double-threaded screws achieved a similar fixation effect as those of bone cement-reinforced screws. Meanwhile, the operation time was shorter, the radiation damage was less, and the difficulty of later revision surgery was reduced. Thus, the proposed surgical protocol can be applied as a new option for patients with osteoporotic lumbar spondylolisthesis.

Interface-driven anomalies in irradiation damage: Multiscale simulation of asymmetric diffusion and mechanical degradation in SnPb solder joints under electron irradiation

Surface-Driven Anomalies in irradiation Damage: Low swelling in 316LN stainless steel under Gold-Ion irradiation up to 290 dpa

Development of a polarized proton solid target for measuring the spin correlation coefficient of deuteron–proton scattering and analysis of radiation damage

High-resolution data and accurate intensity measurements are crucial for resolving detailed structural models in macromolecular electron crystallography. However, obtaining high-quality structural information at atomic resolution is generally difficult, especially for crystalline biological specimens, as the diffracted intensities rapidly fade at higher resolution. Increasing the fluence does not circumvent this problem, as it results in detrimental radiation damage that compromises data quality. The diffraction signal at low flux density conditions is therefore generally expected to be relatively weak, limiting the attainable resolution. Recent advances in macromolecular MicroED data collection have substantially improved data quality, making it possible to extract high-resolution information from small nanocrystals that would otherwise be difficult to analyze. Notably, the introduction of hybrid pixel detectors and direct electron detectors has revolutionized diffraction data acquisition. These cameras essentially have no read-out noise and greatly improve the signal-to-noise ratio, which is particularly beneficial for recording reliable data from faint high- resolution reflections. In many cases, data quality and resolution can further be improved using energy filtering, which decreases background noise and sharpens diffraction peaks by removing inelastically scattered electrons. These technological advances allow for improved data collection strategies, enabling more accurate intensity measurements at higher resolution. Better data leads to better structural models that provide deeper insights into protein structure and function.

Future frontier accelerators envision the use of silicon sensors in environments with fluences exceeding 1 × 1017 1 MeV neq/cm2. Presently available silicon sensors can operate efficiently up to fluences of the order of 1016 1 MeV neq/cm2. Therefore, novel sensors and readout electronics must be developed. Within this framework, state-of-the-art Technology CAD (TCAD) tools can be proficiently used to account for both bulk and surface radiation-induced damage effects in semiconductor sensors, fostering design optimization and enabling a predictive insight into the electrical behaviour of novel solid-state detectors. In particular, the balance between extending already developed models and methodologies or devising different approaches should be carefully considered. In this contribution, the different available TCAD numerical models addressing bulk and surface radiation damage effects will be illustrated. It will also be shown how these models have been used for the optimization of devices, particularly 3D sensors and Low Gain Avalanche Diodes. Moreover, extending the applicability of these models to extreme fluence scenarios requires carefully accounting for the modeling of acceptor and donor removal, impact ionization, carrier mobility and lifetime, and trap dynamics.

TCAD simulations of radiation damage in 4H-SiC

Radiation damage of the W2 plastic scintillator under ultra-high dose rate FLASH electron beam

The macromolecular crystallography (MX) beamline I04 [ref 1] at Diamond has evolved over time through various upgrade projects that aimed at increasing scientific capability but at the same time aiming for increased stability so that the best possible data can be obtained by the user. We have implemented an optical concept for beam delivery combining simplicity with stability. Variable focus beam is provided through the combination of a double crystal monochromator (DCM) with a F-switch which houses compound refractive lenses (CRL) that can be brought individually into the beam path, providing maximum flexibility. Both devices were designed inhouse and this combination allows delivery of a very stable beam from the microfocus regime (8 μm x 5 μm (h x v)) to larger beam sizes (up to 110 μm x 100 μm). Beam delivery within 3% RMS of the beamsize is achieved by making use of a dedicated feedback system using X-ray beam position monitors (XBPMs) [ref 2]. The original X-ray source has been replaced by a 17.6 mm period CPMU in June 2022 and has resulted in a significant flux increase over the whole energy range (6-18 keV) thereby generating new scientific opportunities.Optimal data collection in MX strongly depends on setting up the right data collection parameters which should be defined by the experimental aim or scientific question that is being asked. With high intensity beamlines radiation damage has become the primary limitation and therefore the total exposure of the sample plays a key role. It became soon clear that with the combination of the high variability of the flux profile from the source combined with the large variability of beam sizes, that the concept of exposure per data collection frame is no longer feasible. Therefore, we have implemented the concept of dose aware data collection [ref 3] where the user is given the option to dial a dose per data set (instead of an exposure time per frame) and this dose should of course be compatible with the experimental aim. We use the programme RADDOSE-3D [ref 4, 5] which takes known information from the beamline (energy, flux, beam size) and currently assumes a standard macromolecular crystal which means that the sample only contains lighter elements and combines this information in the dose calculation to produce optimal exposure times per frame and adjustment of transmission if required. Future improvements will consider better information about sample composition and size. In most cases, we aim for the shortest possible exposure time to take advantage of the Eiger2 XE 16M detector capabilities which allows acquisition rates up to 500 Hz. Depending on the experimental aim we usually implement a multi-sweep (and often multi-crystal) approach [ref 6] using different crystal orientations which can be realised with the SmarGon multi-axis goniometer. The dose-based approach is also fully implemented in our unattended data collection (UDC) protocols. Apart from the UDC and remote interactive modes we also strongly encourage in person visits to train users in best practice dose aware data collection and enabling them to make the best choices using the available tools in the data collection software. We constantly aim to streamline the user experience further by addition of new functionality and tools.

X-ray diffraction (XRD) of microcrystals is signal-to-noise limited by the inherently weak diffraction. As such, Electron diffraction (ED) is increasingly used to measure diffraction data from submicron crystals, or those deemed too small for XRD due the stronger interaction of electrons with matter. However, many samples which are too thin for XRD are often too thick for ED using the currently available electron beam energies (<300 keV) and hence require thinning by focussed ion beam milling (FIB) which adds additional sample preparation steps. In addition to determining structures from nanocrystals, ED provides Coulomb potential data which are complementary to that obtained with XRD. As such ED data may be necessary to answer particular scientific questions.The macromolecular crystallography beamline, VMXm, at Diamond Light Source, has been optimised for maximising the S:N in XRD experiments with a variable focus high-energy (>20 KeV) X-ray beam, with in-vacuum endstation and the use of low background cryoTEM grids for crystal mounting [1], [2]. This has allowed VMXm to collect high-resolution rotation data from single crystals measuring ∼1.2 μm which were only previously tractable using an X-ray Free Electron Laser [3]. This has pushed the amenable sample envelope at synchrotrons to new dimensions and perhaps near to the practical limit of XRD. Indeed, simulations have predicted the limit to be ∼0.5 μm thick in the case of lysozyme, assuming photoelectron escape [4]. This XRD beamline opens up the possibilities to directly compare XRD and ED datasets and understand the complementarity of these experiments.In this work we present data from cubic human insulin crystals that have been thinned by FIB milling from ∼10 μm to various submicron thicknesses. 200 kV ED data were then collected from these lamellae before XRD data were measured from the same lamellae using VMXm. It was possible to obtain a complete XRD dataset to 2.45 Å using a 1.68 μm3 illuminated volume and a 2.04 Å ED dataset from the same 0.25 μm lamella. We have demonstrated that the data quality is comparable between ED and VMXm from the same crystal, while giving an opportunity to directly compare X-ray and electron derived maps. This includes the comparison of the radiation damage each experiment imparts on the sample [5] as well as the information content [6]. This work indicates that the usable sample envelope for synchrotron X-rays extends to much thinner samples than had been previously thought. It is also the first demonstration of ED and XRD measured from the same crystal volume enabling direct comparison of X-ray and electron derived data. Ultimately, the work will inform the design and use of high energy (MeV) ED instruments such as HeXI and how those can be complemented by XRD derived information from beamlines such as VMXm.

The Advanced Photon Source (APS) underwent a comprehensive upgrade to replace the original electron storage ring with a new storage ring (APS-U) that increased the X-ray brightness by up to 500 times compared to the APS. The National Institute of General Medical Sciences and National Cancer Institute Structural Biology Facility at the Advanced Photon Source (GM/CA@APS) operates a national user facility for structural biology. During the year-long shutdown for the APS-U upgrade, the GM/CA beamlines and infrastructure were almost completely rebuilt to exploit the high brightness of the APS-U. New state-of-the-art focusing optics (JTECH Corp. mirrors in Axilon AG benders, and RXOPTICS GmbH and Co. compound refractive lenses (CRLs) in AXILON translocators) now provide an extremely intense (>1013 photons/sec), clean, stable, and rapidly adjustable beam size between 1-50 microns. The maximum energy on 23-ID-D was increased to 35 keV to minimize radiation damage. The new high-stability end station table (Axilon AG) supports both the CRL transfocator and sample environment. The new goniometer allows data collection on crystals down to one micron in size and provides capabilities for rapid scanning of random or periodic fixed target samples. The recently acquired DECTRIS Eiger2 XE 16M CdTe detector enables high-speed, high-efficiency X-ray detection on our high- energy beamline 23-ID-D. The new PyBluIce GUI and beamline control software enables sophisticated routines such as 3D-rastering, helical and fully automated (unattended) data collection, and routine serial crystallography data collection from fixed target and injector- based sample delivery systems. Technical beamline commissioning with the first X-rays began on November 22, 2024, along with macromolecular crystallography measurements of standard samples. On March 11, 2025, we began welcoming users back to participate in scientific commissioning. Here, we will present the new designs and game-changing opportunities for structural biology research enabled by these small, ultra-intense, high-energy beams.GM/CA@APS has been funded by the National Cancer Institute (ACB-12002) and the National Institute of General Medical Sciences (AGM-12006, P30GM138396). NIH-Office of Research Infrastructure Programs provided funding for the detectors via High-End Shared Instrumentation Grants (Eiger 16M Si- 1S10OD012289-01A1; and Eiger2 Xe 16M CdTe - S10OD034267). This research used resources of the Advanced Photon Source; a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract No. DE-AC02-06CH11357.