[18F]FSPG-PET provides an early marker of radiotherapy response in head and neck squamous cell cancer.

The ability to image early treatment response to radiotherapy in head and neck squamous cell carcinoma (HNSCC) will enable the identification of radioresistant tumor volumes suitable for treatment intensification. Here, we propose the system xc - radiotracer (4S)-4-(3-[18F]fluoropropyl)-L-glutamate ([18F]FSPG) as a non-invasive method to monitor radiation response in HNSCC. We assessed temporal changes in cell death, antioxidant status, and [18F]FSPG retention following a single dose of 10 Gy irradiation in FaDU HNSCC cells. Next, using a fractionated course of radiotherapy, we assessed tumor volume changes and performed [18F]FSPG-PET imaging in FaDU-bearing mouse xenografts, followed by ex vivo response assessment. In cells, 10 Gy irradiation reduced [18F]FSPG retention, coinciding with the induction of apoptosis and the production of reactive oxygen species. In vivo, [18F]FSPG tumor retention was halved seven days after the start of treatment, which preceded radiotherapy-induced tumor shrinkage, thereby confirming [18F]FSPG-PET as an early and sensitive marker of radiation response.

Census tract-level social vulnerability and gonorrhea rates in central Ohio: results from the STD Surveillance Network, 2020-2021.

BACKGROUND: Gonorrhea remains a significant burden in central Ohio, particularly in socially disadvantaged communities. This study evaluates gonorrhea case rates and changes from 2020 to 2021, focusing on geographic patterns associated with social vulnerability. METHODS: We analyzed gonorrhea case data from the Columbus STD Surveillance Network (SSuN) for 2020 and 2021. Cases were linked to census tracts and categorized into low (0-0.66) or high (0.67-1.0) vulnerability groups based on the CDC/ATSDR social vulnerability index (SVI). We mapped gonorrhea case rates per 100,000 population, stratified by SVI group, and compared changes using Jenks natural breaks to determine cutpoints. High case rates were defined as ≥605 (2020) and ≥ 532 (2021). A change in case rates was defined as <154 (decrease/no change) or ≥ 154 (increase). Demographic factors were summarized for both years. RESULTS: In 2020 and 2021, nearly half of high SVI tracts had high gonorrhea rates, compared to about 4-7% of low SVI tracts. Median rates in high SVI tracts were approximately five times higher than in low SVI tracts, with rates 546 and 598 per 100,000 in 2020 and 2021, compared to 98 and 104 per 100,000, respectively. Gonorrhea rates increased in 23 (9%) of low SVI tracts and 37 (25%) of high SVI tracts. Demographic characteristics remained consistent between the two years. CONCLUSIONS: Gonorrhea rates increased in central Ohio between 2020 and 2021, with higher burdens in high SVI census tracts. Understanding the dynamics of social vulnerability at the community level is crucial for targeting limited STD resources effectively.

A Modular Strategy for the Synthesis of Macrocycles and Medium-Sized Rings via Cyclization/Ring Expansion Cascade Reactions.

Macrocycles and medium-sized rings are important in many scientific fields and technologies but are hard to make using current methods, especially on a large scale. Outlined herein is a strategy by which functionalized macrocycles and medium-sized rings can be prepared using cyclization/ring expansion (CRE) cascade reactions, without resorting to high dilution conditions. CRE cascade reactions are designed to operate exclusively via kinetically favorable 5-7-membered ring cyclization steps; this means that the problems typically associated with classical end-to-end macrocyclization reactions are avoided. A modular synthetic approach has been developed to facilitate the simple assembly of the requisite linear precursors, which can then be converted into an extremely broad range of functionalized macrocycles and medium-sized rings using one of nine CRE protocols.

Divergent Cascade Ring-Expansion Reactions of Acryloyl Imides.

Macrocyclic and medium-sized ring ketones, lactones and lactams can all be made from common acryloyl imide starting materials through divergent, one-pot cascade ring-expansion reactions. Following either conjugate addition with an amine or nitromethane, or osmium(VIII)-catalysed dihydoxylation, rearrangement through a four-atom ring expansion takes place spontaneously to form the ring expanded products. A second ring expansion can also be performed following a second iteration of imide formation and alkene functionalisation/ring expansion. In the dihydroxylation series, three- or four-atom ring expansion can be performed selectively, depending on whether the reaction is under kinetic or thermodynamic control.

DamID-seq: A Genome-Wide DNA Methylation Method that Captures Both Transient and Stable TF-DNA Interactions in Plant Cells.

Capturing the dynamic and transient interactions of a transcription factor (TF) with its genome-wide targets whose regulation leads to plants' adaptation to their changing environment is a major technical challenge. This is a widespread problem with biochemical methods such as chromatin immunoprecipitation-sequencing (ChIP-seq) which are biased towards capturing stable TF-target gene interactions. Herein, we describe how DNA adenine methyltransferase identification and sequencing (DamID-seq) can be used to capture both transient and stable TF-target interactions by DNA methylation. The DamID technique uses a TF protein fused to a DNA adenine methyltransferase (Dam) from E. coli. When expressed in a plant cell, the Dam-TF fusion protein will methylate adenine (A) bases near the sites of TF-DNA interactions. In this way, DamID results in a permanent, stable DNA methylation mark on TF-target gene promoters, even if the target gene is only transiently "touched" by the Dam-TF fusion protein. Here we provide a step-by-step protocol to perform DamID-seq experiments in isolated plant cells for any Dam-TF fusion protein of interest. We also provide information that will enable researchers to analyze DamID-seq data to identify TF-binding sites in the genome. Our protocol includes instructions for vector cloning of the Dam-TF fusion proteins, plant cell protoplast transfections, DamID preps, library preparation, and sequencing data analysis. The protocol outlined in this chapter is performed in Arabidopsis thaliana, however, the DamID-seq workflow developed in this guide is broadly applicable to other plants and organisms.

Double DAP-seq uncovered synergistic DNA binding of interacting bZIP transcription factors.

Many eukaryotic transcription factors (TF) form homodimer or heterodimer complexes to regulate gene expression. Dimerization of BASIC LEUCINE ZIPPER (bZIP) TFs are critical for their functions, but the molecular mechanism underlying the DNA binding and functional specificity of homo- versus heterodimers remains elusive. To address this gap, we present the double DNA Affinity Purification-sequencing (dDAP-seq) technique that maps heterodimer binding sites on endogenous genomic DNA. Using dDAP-seq we profile twenty pairs of C/S1 bZIP heterodimers and S1 homodimers in Arabidopsis and show that heterodimerization significantly expands the DNA binding preferences of these TFs. Analysis of dDAP-seq binding sites reveals the function of bZIP9 in abscisic acid response and the role of bZIP53 heterodimer-specific binding in seed maturation. The C/S1 heterodimers show distinct preferences for the ACGT elements recognized by plant bZIPs and motifs resembling the yeast GCN4 cis-elements. This study demonstrates the potential of dDAP-seq in deciphering the DNA binding specificities of interacting TFs that are key for combinatorial gene regulation.

The chicken chorioallantoic membrane as a low-cost, high-throughput model for cancer imaging.

Mouse models are invaluable tools for radiotracer development and validation. They are, however, expensive, low throughput, and are constrained by animal welfare considerations. Here, we assessed the chicken chorioallantoic membrane (CAM) as an alternative to mice for preclinical cancer imaging studies. NCI-H460 FLuc cells grown in Matrigel on the CAM formed vascularized tumors of reproducible size without compromising embryo viability. By designing a simple method for vessel cannulation it was possible to perform dynamic PET imaging in ovo, producing high tumor-to-background signal for both 18F-2-fluoro-2-deoxy-D-glucose (18F-FDG) and (4S)-4-(3-18F-fluoropropyl)-L-glutamate (18F-FSPG). The pattern of 18F-FDG tumor uptake were similar in ovo and in vivo, although tumor-associated radioactivity was higher in the CAM-grown tumors over the 60 min imaging time course. Additionally, 18F-FSPG provided an early marker of both treatment response to external beam radiotherapy and target inhibition in ovo. Overall, the CAM provided a low-cost alternative to tumor xenograft mouse models which may broaden access to PET and SPECT imaging and have utility across multiple applications.

Imaging the master regulator of the antioxidant response in non-small cell lung cancer with positron emission tomography.

Mutations in the NRF2-KEAP1 pathway are common in non-small cell lung cancer (NSCLC) and confer broad-spectrum therapeutic resistance, leading to poor outcomes. The cystine/glutamate antiporter, system xc-, is one of the >200 cytoprotective proteins controlled by NRF2, which can be non-invasively imaged by (S)-4-(3-18F-fluoropropyl)-l-glutamate ([18F]FSPG) positron emission tomography (PET). Through genetic and pharmacologic manipulation, we show that [18F]FSPG provides a sensitive and specific marker of NRF2 activation in advanced preclinical models of NSCLC. We validate imaging readouts with metabolomic measurements of system xc- activity and their coupling to intracellular glutathione concentration. A redox gene signature was measured in patients from the TRACERx 421 cohort, suggesting an opportunity for patient stratification prior to imaging. Furthermore, we reveal that system xc- is a metabolic vulnerability that can be therapeutically targeted for sustained tumour growth suppression in aggressive NSCLC. Our results establish [18F]FSPG as predictive marker of therapy resistance in NSCLC and provide the basis for the clinical evaluation of both imaging and therapeutic agents that target this important antioxidant pathway.

Alone with my phone? Examining beliefs about solitude and technology use in adolescence.

In this study, we examined how technology impacts adolescents' perceptions of, and affective responses to solitude, as well as how adolescents' own motivations for solitude (shyness, affinity for aloneness) were related to these reactions. Participants were N = 437 adolescents (297 girls; M age = 16.15 years, standard deviation (SD) = .50) who were presented with a series of hypothetical vignettes asking them to imagine themselves in the context of pure solitude (alone in their room with the door closed), as well as being physically alone but engaged in increasing levels of virtual social engagement, including passive (e.g., watching videos, scrolling, but no direct social engagement), active (e.g., texting), and audio-visual (e.g., Facetime) technology use. Following each vignette, participants reported their perceptions of being alone and positive/negative affective responses. We also measured general motivations for solitude (shyness, affinity for aloneness). Among the results, adolescents perceived themselves as less alone in vignettes depicting increasing virtual social engagement. Affective benefits of increased virtual engagement were also found (e.g., less loneliness/boredom/sadness, greater social connection/contentment). However, these effects were moderated by solitude motivations, with different patterns evident as a function of participant shyness and affinity for aloneness. Findings highlight the importance of considering the nature of adolescents' technology use when alone, as well as motivations for solitude, when considering links between solitude and well-being.

Crystal structures of three zinc(II) halide coordination complexes with quinoline N-oxide.

The reaction of one equivalent of zinc(II) halide with two equivalents of quinoline N-oxide (QNO) in methanol yields compounds as ZnX 2(QNO)2, where X = Cl (I), Br (II) and I (III), namely, di-chlorido-bis-(quinoline N-oxide-κO)zinc(II), [ZnCl2(C9H7NO)2], di-bromido-bis-(quinoline N-oxide-κO)zinc(II), [ZnBr2(C9H7NO)2], and di-iodido-bis-(quinoline N-oxide-κO)zinc(II) [ZnI2(C9H7NO)2]. In all three complexes, Zn cations are coordinated by two QNO ligands bound through the oxygen atoms and two halide atoms, with X-Zn-X bond angles ca 20° wider than the O-Zn-O, giving rise to a distorted tetra-hedral geometry. Crystals of (II) and (III) are isostructural and both show pairwise π-stacking of QNO ligands and weak C-H⋯X hydrogen bonds, while (I) packs differently, with a shorter C-H⋯Cl bond and without π-stacking.

Comparative Analysis of Transcriptomes of Ophiostoma novo-ulmi ssp. americana Colonizing Resistant or Sensitive Genotypes of American Elm.

The Ascomycete Ophiostoma novo-ulmi threatens elm populations worldwide. The molecular mechanisms underlying its pathogenicity and virulence are still largely uncharacterized. As part of a collaborative study of the O. novo-ulmi-elm interactome, we analyzed the O. novo-ulmi ssp. americana transcriptomes obtained by deep sequencing of messenger RNAs recovered from Ulmus americana saplings from one resistant (Valley Forge, VF) and one susceptible (S) elm genotypes at 0 and 96 h post-inoculation (hpi). Transcripts were identified for 6424 of the 8640 protein-coding genes annotated in the O. novo-ulmi nuclear genome. A total of 1439 genes expressed in planta had orthologs in the PHI-base curated database of genes involved in host-pathogen interactions, whereas 472 genes were considered differentially expressed (DEG) in S elms (370 genes) and VF elms (102 genes) at 96 hpi. Gene ontology (GO) terms for processes and activities associated with transport and transmembrane transport accounted for half (27/55) of GO terms that were significantly enriched in fungal genes upregulated in S elms, whereas the 22 GO terms enriched in genes overexpressed in VF elms included nine GO terms associated with metabolism, catabolism and transport of carbohydrates. Weighted gene co-expression network analysis identified three modules that were significantly associated with higher gene expression in S elms. The three modules accounted for 727 genes expressed in planta and included 103 DEGs upregulated in S elms. Knockdown- and knockout mutants were obtained for eight O. novo-ulmi genes. Although mutants remained virulent towards U. americana saplings, we identified a large repertoire of additional candidate O. novo-ulmi pathogenicity genes for functional validation by loss-of-function approaches.

Normative Beliefs about Social Withdrawal in Adolescence.

The goal of this study is to explore normative beliefs about social withdrawal during adolescence. Participants were N = 419 adolescents (Mage = 16.13 years), who completed measures of normative beliefs about social withdrawal and their own social withdrawal (shyness, unsociability). Among the results, adolescents reported greater overall acceptance of unsociability compared to shyness, however, some gender differences also emerged. Specifically, adolescents were more accepting of unsociability when depicted by hypothetical females, and more accepting of shyness when depicted by hypothetical males. Participant social withdrawal was associated with greater acceptance of socially withdrawn behaviors in others. These findings provide a first look at normative beliefs about social withdrawal during a developmentally critical age period for this construct.

Emotion dynamics in movie dialogues.

Emotion dynamics is a framework for measuring how an individual's emotions change over time. It is a powerful tool for understanding how we behave and interact with the world. In this paper, we introduce a framework to track emotion dynamics through one's utterances. Specifically we introduce a number of utterance emotion dynamics (UED) metrics inspired by work in Psychology. We use this approach to trace emotional arcs of movie characters. We analyze thousands of such character arcs to test hypotheses that inform our broader understanding of stories. Notably, we show that there is a tendency for characters to use increasingly more negative words and become increasingly emotionally discordant with each other until about 90% of the narrative length. UED also has applications in behavior studies, social sciences, and public health.

Feasibility of a syndrome-informed micro-intervention for infants with Down syndrome.

BACKGROUND: Infants with Down syndrome (DS) are at risk for a range of phenotypic outcomes, including delays in the onset of reaching behaviour, a critical skill that facilitates early learning. This parallel-group feasibility and pilot study presents findings from a parent-mediated micro-intervention that aimed to support the development of reaching behaviour in a sample of infants with DS. METHODS: Participants were 73 infants with DS and their caregivers. Infants who qualified for the home-based intervention (based on manual skill performance on Bayley Scales of Infant and Toddler Development, Third Edition items) were randomly assigned individually or by geographical region to a treatment or an alternative treatment condition that involved toy-based interactions with caregivers. Infants in the treatment condition experienced facilitated reaching during the toy-based interactions through the use of Velcro-affixed mittens and toys. RESULTS: Forty-two infants met criteria to participate in the intervention, and 37 participated in both baseline and post-treatment visits. At post-treatment, infants in the treatment condition demonstrated shorter latencies to make contact with objects and showed higher frequencies of reach attempts and swats at objects than infants in the alternative treatment group. These findings were more pronounced when examining a chronological age-restricted subgroup of infants 5 to 10 months. CONCLUSIONS: Findings suggest that a syndrome-informed approach to targeted intervention may be a promising application of phenotyping science in DS and other neurogenetic conditions associated with intellectual disability.

Social Withdrawal and Aloneliness in Adolescence: Examining the Implications of Too Much and Not Enough Solitude.

Adolescence is a developmental period when spending time alone becomes particularly important. The aim of this study was to explore adolescents' experiences of solitude, with a focus on the implications of both spending too much time alone-as well as not enough time alone. Participants were N = 869 adolescents (15-19 years of age, M = 16.14, SD = 0.50), who completed a series of self report measures assessing time spent alone, activities while alone, social motivations (shyness, sociability), affect, and the recently introduced construct of aloneliness (i.e., negative feeling that arise from the perception that one is not getting enough time in solitude). Among the results, person-oriented analyses provided evidence of distinct subgroups of socially withdrawn adolescents (e.g., shy-withdrawn, unsociable, socially avoidant) as well as an alonely group (high aloneliness, low time alone). There was also preliminary support for the notion that solitary activity activities that are intrinsically motivated may be particularly effective at satisfying the need for solitude (and thereby reducing feelings of aloneliness). Taken together, these results highlight the importance of considering the implications of getting both too much-as well as not enough solitude for adolescents' well-being.

Gene expression changes occurring at bolting time are associated with leaf senescence in Arabidopsis.

In plants, the vegetative to reproductive phase transition (termed bolting in Arabidopsis) generally precedes age-dependent leaf senescence (LS). Many studies describe a temporal link between bolting time and LS, as plants that bolt early, senesce early, and plants that bolt late, senesce late. The molecular mechanisms underlying this relationship are unknown and are potentially agriculturally important, as they may allow for the development of crops that can overcome early LS caused by stress-related early-phase transition. We hypothesized that leaf gene expression changes occurring in synchrony with bolting were regulating LS. ARABIDOPSIS TRITHORAX (ATX) enzymes are general methyltransferases that regulate the adult vegetative to reproductive phase transition. We generated an atx1, atx3, and atx4 (atx1,3,4) triple T-DNA insertion mutant that displays both early bolting and early LS. This mutant was used in an RNA-seq time-series experiment to identify gene expression changes in rosette leaves that are likely associated with bolting. By comparing the early bolting mutant to vegetative WT plants of the same age, we were able to generate a list of differentially expressed genes (DEGs) that change expression with bolting as the plants age. We trimmed the list by intersection with publicly available WT datasets, which removed genes from our DEG list that were atx1,3,4 specific. The resulting 398 bolting-associated genes (BAGs) are differentially expressed in a mature rosette leaf at bolting. The BAG list contains many well-characterized LS regulators (ORE1, WRKY45, NAP, WRKY28), and GO analysis revealed enrichment for LS and LS-related processes. These bolting-associated LS regulators may contribute to the temporal coupling of bolting time to LS.

Does the Presence of Chondral Lesions Negatively Affect Patient-Determined Outcomes After Arthroscopic Rotator Cuff Repair?

BACKGROUND: There are limited data available to guide patients to their prognosis when glenohumeral chondral lesions are found during arthroscopic rotator cuff repair. HYPOTHESIS: The primary hypothesis was that patients with glenohumeral chondral lesions will have inferior outcomes after arthroscopic rotator cuff repair compared with patients without chondral lesions. The secondary hypothesis was that patients with concomitant chondral lesions will have more severe preoperative symptoms compared with those without chondral lesions. STUDY DESIGN: Cohort study; Level of evidence, 3. METHODS: A retrospective analysis was performed of patients who underwent arthroscopic rotator cuff repair between 2008 and 2012. We examined the effects of chondral lesions on patient-determined outcomes, which included the Western Ontario Rotator Cuff Index (WORC), American Shoulder and Elbow Surgeons (ASES) score, Simple Shoulder Test (SST), Single Assessment Numeric Evaluation (SANE), and the Shoulder Activity Level (SAL). Shoulders without chondral lesions were compared with shoulders with chondral lesions to determine whether differences in severity of preoperative symptoms as well as postoperative improvements were statistically significant. RESULTS: A total of 281 shoulders were included from 273 patients, with a mean follow-up of 3.7 years. In total, 90 shoulders (32%) had concomitant chondral lesions in the glenohumeral joint. The presence and degree of chondral damage were not associated with the severity of preoperative symptoms or the amount of improvement after arthroscopic rotator cuff repair, as determined by patient outcome scores. Shoulders with bipolar chondral lesions had less postoperative improvement in their outcome scores compared with shoulders with unipolar lesions, with significant differences found in the SST (P = .0005), the SANE (P = .005), and the SAL (P = .04). Regardless of this, the majority of shoulders with bipolar chondral lesions (80%-92%) had postoperative improvements that superseded the minimal clinically important difference of the ASES, WORC, and SANE. CONCLUSION: At a mean 3.7-year follow-up, the presence of chondral damage did not appear to negatively affect the improvement in patient-determined outcomes after arthroscopic rotator cuff repair. However, improvement in outcomes was negatively affected by the presence of bipolar chondral lesions.

Do patients who smoke tobacco have success with primary arthroscopic rotator cuff repair? A comparison with nonsmokers.

BACKGROUND: It is recommended that patients cease smoking before rotator cuff repair. However, not all patients want to or are able to successfully cease smoking. This raises the question if these patients should be advised to pursue surgical intervention or if surgery should be contraindicated until patients successfully cease smoking. METHODS: A retrospective analysis of patients undergoing rotator cuff repair was performed to examine the effects of smoking tobacco on patient-determined outcomes (Western Ontario Rotator Cuff Index [WORC], American Shoulder and Elbow Surgeons score [ASES], Simple Shoulder Test [SST], and Single Assessment Numeric Evaluation [SANE]). Patients who smoked tobacco at the time of surgery were compared with patients who were not smoking to determine if differences in (1) severity of preoperative and postoperative symptoms and (2) the postoperative improvements were statistically significant. RESULTS: Thirty-one patients were smokers and 205 were nonsmokers. Preoperative scores were worse for smokers compared with nonsmokers: WORC (32 vs. 43; P = .0002), ASES (32 vs. 43; P = .001), SST (3.5 vs. 4.6; P = .04), and SANE (34 vs. 38; P = .35). Postoperative scores were worse for smokers compared with nonsmokers: WORC (79 vs. 89; P = .001), ASES (82 vs. 89; P = .04), SST (9.0 vs. 10.2; P = .02), and SANE (84 vs. 89; P = .09). There were no significant differences in change in scores over time or percentage of patients achieving the minimal clinically important difference of the score between groups. CONCLUSIONS: From examining the patients' subjective patient-determined outcome scores, it does not appear that rotator cuff repair should be strictly contraindicated in active smokers. Postoperative improvements in smokers were similar to nonsmokers. Smokers have lower baseline preoperative and postoperative outcome scores compared with nonsmokers.

Ethyl 1H-indole-2-carboxyl-ate.

Our work in the area of synthesis of tris indole compounds as a potential chelator led to the synthesis and crystallization of ethyl 1H-indole-2-carboxyl-ate, C11H11NO2, an indole that was synthesized by the thionyl chloride reaction of 1H-indole-2-carb-oxy-lic acid, followed by dissolution in ethanol. The mol-ecular packing exhibits a herringbone pattern with the zigzag running along the b-axis direction; the compound crystallizes as a hydrogen-bonded dimer resulting from O⋯H-N hydrogen bonds, between the indole N-H group and the keto oxygen atom, which build centrosymmetric R 2 2(10) ring motifs in the crystal.

Pyridine-4-carboxamidoxime N-oxide.

Our work in the area of synthesis of metal-organic frameworks (MOFs) based on organic N-oxides led to the crystallization of pyridine-4-carboxamidoxime N-oxide. Herein we report the first crystal structure of the title compound, C6H7N3O2 [systematic name: (Z)-4-(N'-hy-droxy-carbamimido-yl)pyridine N-oxide]. The hy-droxy-carbamimidoyl group is essentially coplanar with the aromatic ring, r.m.s.d. = 0.112 Å. The compound crystallizes in hydrogen-bonding layers built from the formation of strong O-H⋯O hydrogen bonds between the oxime oxygen atom and the oxygen atom of the N-oxide, and the formation of N-H⋯O hydrogen bonds between one amine nitro-gen atom and the N-oxide oxygen atom. These combined build R 3 4(24) ring motifs in the crystal. The crystal structure has no π-π inter-actions.