Singlet oxygen (1O2) is the primary cytotoxic agent in Type-II photodynamic therapy (PDT). Single-photon avalanche diode (SPAD)-based time-resolved singlet oxygen luminescence detection (TSOLD) systems enable direct detection of the near-infrared 1O2 phosphorescence (∼1270 nm) during PDT, offering a powerful tool for treatment monitoring. However, the efficiency of detection depends strongly on the acquisition parameters. Here, we present a comprehensive evaluation of a TSOLD system with 630 nm and 690 nm nanosecond pulsed diode lasers developed for in vivo PDT monitoring. The influence of acquisition time (1-300 s), SPAD dead time (5-40 µs), and temporal bin width (1-100 ns) on the fidelity of 1O2 lifetime measurements and signal-to-noise ratio (SNR) was quantitatively investigated. Measurements were performed using liquid phantoms containing the photosensitizers Photofrin or benzoporphyrin derivative, with acquisition time further validated in vivo in murine tumor models. The results demonstrate that both acquisition time and detector dead time significantly affect the signal vs. time histogram shape and the accuracy of the 1O2 and photosensitizer triplet-state lifetime measurements, while an optimal bin width minimizes photon-count noise without compromising temporal resolution. The optimized parameters enable reliable 1O2 lifetime extraction within 100 s from the mouse model in vivo. This systematic evaluation establishes quantitative design guidelines for compact TSOLD systems tailored to in vivo applications.

Circular RNAs (circRNAs) are a class of endogenous non-coding RNAs characterized by a covalently closed-loop structure. Although initially regarded as splicing byproducts, circRNAs are now recognized as essential regulators of gene and protein expression and play important roles in various human diseases. In recent years, growing evidence has indicated that a subset of circRNAs can be translated through cap-independent mechanisms to produce bioactive peptides or proteins. These findings expand the functional scope of circRNAs and offer new opportunities for RNA therapy. This review summarizes recent advances in circRNA biology, with an emphasis on their potential in diagnosis and treatment. We also review the therapeutic strategies targeting circRNAs at both the RNA and protein levels and the delivery systems that support circRNA-directed therapies. After we discuss how circRNA therapeutics may be integrated into precision medicine, we further highlight the current clinical progress and key challenges for advancing circRNA-based therapies toward clinical application.

Although machine learning has transformed protein structure prediction of folded protein ground states with remarkable accuracy, intrinsically disordered proteins and regions (IDPs/IDRs) are defined by diverse and dynamical structural ensembles that are predicted with low confidence by algorithms such as AlphaFold and RoseTTAFold. We present a new machine learning method, IDPForge (IntrinsicallyDisorderedProtein,FOlded and disorderedRegionGEnerator), that exploits a transformer protein language diffusion model to create all-atom IDP ensembles and IDR disordered ensembles that maintains the folded domains. IDPForge does not require sequence-specific training, back transformations from coarse-grained representations, nor ensemble reweighting, as in general the created IDP/IDR conformational ensembles show good agreement with solution experimental data, and options for biasing with experimental restraints are provided if desired. We envision that IDPForge with these diverse capabilities will facilitate integrative and structural studies for proteins that contain intrinsic disorder, and is available as an open source resource for general use.

Not available.

Abstract Measuring the ionizing photon production efficiency ξ ion,0 —the ratio of the ionizing photon output rate <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" overflow="scroll"> <mml:msub> <mml:mrow> <mml:mi>Q</mml:mi> </mml:mrow> <mml:mrow> <mml:msup> <mml:mrow> <mml:mi mathvariant="normal">H</mml:mi> </mml:mrow> <mml:mrow> <mml:mn>0</mml:mn> </mml:mrow> </mml:msup> </mml:mrow> </mml:msub> </mml:math> to the UV continuum luminosity L UV —in galaxies at z &gt; 6 is crucial for constraining their contribution to cosmic reionization. We present integrated and spatially resolved measurements of ξ ion,0 for 12 exceptionally bright ( M UV ∼ −22 mag) star-forming galaxies at z ∼ 7 from the Reionization Era Bright Emission Line Survey (REBELS). These measurements are based on JWST NIRSpec/Integral Field Unit PRISM spectroscopy, probing the rest-frame UV and optical regime. Notably, in 8 of the 12 galaxies, the spectral coverage includes H α , enabling self-consistent dust attenuation estimates in both the ionized gas and stellar continuum via the Balmer decrement and rest-UV slope, respectively. We find global <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" overflow="scroll"> <mml:mi>log</mml:mi> <mml:mo stretchy="false">(</mml:mo> <mml:msub> <mml:mrow> <mml:mi>ξ</mml:mi> </mml:mrow> <mml:mrow> <mml:mi mathvariant="normal">ion,0</mml:mi> </mml:mrow> </mml:msub> <mml:mo stretchy="false">)</mml:mo> </mml:math> values ranging from 25.19 ± 0.11 to 25.61 ± 0.11, with a weighted mean of 25.44 ± 0.15, consistent with the canonical value of ∼25.3. Using a sample of 25 star-forming clumps within these galaxies, we explore local variations in the ionizing photon production efficiency, finding a broader range, from 24.52 ± 0.21 to 26.18 ± 0.61. We identify strong correlations between ξ ion,0 and the specific star formation rate, star formation surface density, H β equivalent width, and stellar mass. Clumps with the highest ξ ion,0 exhibit EW 0 (H β ) ≥ 150 Å, consistent with young stellar ages. From previous Ly α measurements in three galaxies, we estimate a typical Ly α escape fraction of f esc,Ly α ∼ 2%, suggesting similar or lower escape fractions for LyC photons. Combining this with our H α measurements, we infer ionized bubble sizes ∼1 pMpc, aligned with expectations from Ly α -detected systems and reionization models.

Functional hyperemia is a well-established hallmark of healthy brain function, whereby local brain blood flow adjusts in response to a change in the activity of the surrounding neurons. Although functional hyperemia has been extensively studied at the level of both tissue and individual vessels, vascular network-level coordination remains largely unknown. To bridge this gap, we developed a deep learning-based pipeline that uses two-photon fluorescence microscopy images of cerebral microcirculation to enable automated reconstruction and quantification of the geometric changes across the microvascular network, comprising hundreds of interconnected blood vessels, pre and post-activation of the neighboring neurons. The pipeline's utility was demonstrated in the Thy1-ChR2 optogenetic mouse model, where we observed network-wide vessel radius changes to depend on the photostimulation intensity, with both dilations and constrictions occurring across the cortical depth, at an average of 16.1±14.3 μm (mean ± SD) away from the most proximal neuron for dilations; and at 21.9±14.6 μm away for constrictions. We observed a significant heterogeneity of the vascular radius changes within vessels, with radius adjustment varying by an average of 24 ± 28% of the resting diameter, likely reflecting the heterogeneity of the distribution of contractile cells on the vessel walls. A graph theory-based network analysis revealed that the assortativity of adjacent blood vessel responses rose by 152 ± 65% at 4.3 mW/mm2 of blue photostimulation vs. the control, with a 4% median increase in the efficiency of the capillary networks during this level of blue photostimulation in relation to the baseline. Interrogating individual vessels is thus not sufficient to predict how the blood flow is modulated in the network. Our pipeline, enables tracking of the microvascular network geometry over time, relating caliber adjustments to vessel wall-associated cells' state, and mapping network-level flow distribution impairments in experimental models of disease.

Traumatic injury, stenosis, and malignancy involving large segments of the airway are difficult to reconstruct and require novel solutions. Despite advances in surgical techniques, the reconstruction of long-segment tracheal defects remains a significant challenge. Several bioengineering approaches have been explored for tracheal regeneration in vitro and in vivo, using cells in combination with three dimentional (3D) biological or synthetic scaffolds. This paper reviews recent advances in developing bioengineered trachea and the technologies utilized toward generating transplantable tracheal grafts. Specifically, the review will focus on the recellularization of tissue-engineered grafts using natural or synthetic scaffolds, highlighting relevant cell types used to reconstitute tracheal epithelium and cartilage. The promise of newly explored paradigms, including the application of pluripotent stem cells, will be discussed with an overview of associated challenges and necessary steps for future translation. Overall, these advances provide a foundation for the development of clinically viable tracheal grafts, bringing engineered tracheal reconstruction closer to reality.

Abstract Glioblastoma (GBM) is an aggressive brain cancer characterized by profound immunosuppression, largely driven by tumor-associated macrophages (TAMs), which account for 30-40% of the tumour bulk. In normal tissue homeostasis, macrophages detect, engulf, and clear stressed and cancerous cells through phagocytosis. TAMs, however, display markedly impaired ability to do so, and instead promote tumour growth. We sought to identify molecular pathways that regulate macrophage phagocytosis of GBM cells, with the goal of reprogramming TAMs into anti-tumor effectors. We performed a pooled genome-wide CRISPR knockout screen in BV2 macrophages. Mutagenized macrophages were co-cultured with murine glioma cells expressing ZsGreen, a fluorescent reporter. Upon phagocytosis, macrophages engulf ZsGreen and thus fluoresce, allowing for separation and collection of phagocytic and non-phagocytic macrophages by flow cytometry. Synthetic guide RNAs (sgRNAs) were recovered and sequenced to identify gene knockouts that enhanced phagocytosis. Analysis of sgRNA enrichment in ZsGreen+ macrophages revealed overrepresentation of gene knockouts involved in glycosaminoglycan biosynthesis, particularly the chondroitin sulfate pathway. Targeted disruption of CS biosynthesis genes in follow-up CRISPR knockout macrophage cell lines increased uptake of GBM cells in vitro, confirming CS synthesis loss enhances tumor cell phagocytosis. To contextualize these findings in GBM, we characterized O-glycan synthesis in patient tumors. We found CS synthesis gene programs are upregulated in GBM compared to normal brain tissue, and myeloid cells are the dominant producers of CS within the GBM niche. These data identify CS synthesis as a novel inhibitory pathway of macrophage phagocytic function in GBM. This work highlights glycosaminoglycan metabolism as a potential target for therapeutic reprogramming of TAMs to enhance innate immunity and immunotherapy efficacy in GBM. Citation Format: Shan Grewal, Vaseem Shaikh, Adrian Granda Farias, Kevin R. Brown, Lucas Asselstine, Mohamed Taleb, Chitra Venugopal, Patricia Mero, Jason Moffat, Sheila K. Singh. Genome-wide CRISPR screening identifies novel regulators of macrophage phagocytosis against glioblastoma [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Brain Cancer; 2026 Mar 23-25; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2026;86(6_Suppl):Abstract nr A011.

Abstract Glioblastoma (GBM) evolves within a microenvironment abundant in oligodendrocyte-lineage (OL) cells. In this study, we utilized single-cell and spatial transcriptomics from primary and recurrent GBM tumors, immunohistochemistry, cytokine profiling, and migration assays to show that GBM cells recruit OLs to the tumor border via CX3CL1/CX3CR1 signaling. A pan-disease human OL meta-atlas and syngeneic mouse models reveal an interferon-induced reactive OL state, akin to those seen in demyelinating inflammatory and traumatic injury, which is enriched in CNS malignancies. These reactive OLs secrete pro-tumorigenic cytokines, notably CCL5, that promote GBM tumor cell growth through CCR5 signaling. CCR5 is preferentially expressed in glioma stem-like cells (GSCs) and upregulated at recurrence. Targeting CCR5 with genetic knockdown or FDA-approved drug Maraviroc impairs GSC stemness and prolongs survival in GBM models. Our work highlights the functional interplay between OLs and GBM cells and positions the CCL5/CCR5 axis as a druggable target in GBM. Citation Format: Nicholas Mikolajewicz, Kui Zhai, Anish Puri, Petar Miltec, Nazanin Tatari, Jiarun Wei, Neil Savage, Zhi Huang, Qian Huang, Seon Yong Lee, Mahta Jan-Ahmadnejad, Roseanne Nguyen, David Chen, Tiegan Korman, Daniel Mobilio, Maxwell Topley, Jack Qinyu Lu, Matthew R. Voisin, Zsolt Zador, Shawn C. Chafe, Chitra Venugopal, Kevin R. Brown, Gelareh Zadeh, Hong Han, Julien Muffat, Shideng Bao, Sheila K. Singh, Jason Moffat. Reactive oligodendrocytes promote glioblastoma progression through CCL5/CCR5-mediated glioma stem cell maintenance [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Brain Cancer; 2026 Mar 23-25; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2026;86(6_Suppl):Abstract nr A040.

Heart failure (HF) due to left ventricular systolic dysfunction (LVSD) remains a major clinical challenge, particularly among patients with chronic total occlusions (CTO). CTO are present in up to 30% of patients with LVSD undergoing coronary angiography and are independently associated with worse outcomes. Although advances in interventional techniques have increased success rates of CTO percutaneous coronary intervention (CTO-PCI), high-quality evidence supporting this procedure in patients with LVSD remains limited. Observational studies report potential benefits, including improved survival, alleviation of HF symptoms, and recovery of left ventricular ejection fraction (LVEF). However, randomized controlled trials (RCTs) have largely excluded patients with LVEF <35% and those with advanced, complex coronary artery disease (CAD), including CTO, thereby restricting generalizability. Assessment of myocardial viability remains central to patient select for CTO-PCI, its prognostic value for hard clinical endpoints has not been definitively established. The use of mechanical circulatory support (MCS) during high-risk CTO-PCI is increasing, particularly in patients with reduced LVEF and complex coronary anatomy; available data provides inconsistent evidence regarding its impact on clinical outcomes and appears to be largely influenced by individual patient characteristics. Finally, in the setting of acute coronary syndromes (ACS), the effect of CTO revascularization on clinical endpoints and arrhythmic risk is unclear, with conflicting observational data. Future research should prioritize this underrepresented high-risk cohort and be conducted in experienced centers within an integrated multidisciplinary care framework.