Single-cell RNA sequencing of pediatric Hodgkin lymphoma to study the inhibition of T cell subtypes.

Pediatric classic Hodgkin lymphoma (cHL) patients have a high survival rate but suffer from severe long-term side effects induced by chemo- and radiotherapy. cHL tumors are characterized by the low fraction (0.1%-10%) of malignant Hodgkin and Reed-Sternberg (HRS) cells in the tumor. The HRS cells depend on the surrounding immune cells for survival and growth. This dependence is leveraged by current treatments that target the PD-1/PD-L1 axis in cHL tumors. The development of more targeted therapies that are specific for the tumor and are therefore less toxic for healthy tissue compared with conventional chemotherapy could improve the quality of life of pediatric cHL survivors. Here, we applied single-cell RNA sequencing (scRNA-seq) on isolated HRS cells and the immune cells from the same cHL tumors. Besides TNFRSF8 (CD30), we identified other genes of cell surface proteins that are consistently overexpressed in HRS cells, such as NRXN3 and LRP8, which can potentially be used as alternative targets for antibody-drug conjugates or CAR T cells. Finally, we identified potential interactions by which HRS cells inhibit T cells, among which are the galectin-1/CD69 and HLA-II/LAG3 interactions. RNAscope was used to validate the enrichment of CD69 and LAG3 expression on T cells near HRS cells and indicated large variability of the interaction strength with the corresponding ligands between patients and between tumor tissue regions. In conclusion, this study identifies new potential therapeutic targets for cHL and highlights the importance of studying heterogeneity when identifying therapy targets, specifically those that target tumor-immune cell interactions.

Discharge with Home Health Care is Associated with Increased Complications, Readmission, and Mortality Following Total Shoulder Arthroplasty.

INTRODUCTION: Patients undergoing primary anatomic and reverse total shoulder arthroplasty (TSA) are often discharged with home health care (HHC) to provide access to at-home services and facilitate postoperative recovery and continued medical management. The purpose of this study is to evaluate the short-term postoperative outcomes of patients following primary TSA discharged with HHC, including medical and surgical complications, total cost of care, and total hospital length of stay (LOS). METHODS: The Nationwide Readmissions Database (NRD) was reviewed for patients who underwent elective primary TSA between 2016 to 2020 for a retrospective cohort analysis. Patients were stratified by discharge status following the inpatient admission, with 32,497 patients discharged with HHC and 116,402 patients discharged routinely with self-care. Patient demographics, preoperative medical comorbidities, postoperative medical and surgical complications within 180 days, cost of admission, and total hospital length of stay (LOS) were compared between the two discharge groups using Chi-squared analyses. Further multivariate analysis was conducted to control for independent prognosticators on the effect of HHC on postoperative outcomes. RESULTS: Discharge with HHC was correlated with significantly increased rates of all-cause medical complications (OR 1.6, p < 0.001), surgical site infection (SSI) (OR 2.8, p < 0.001), hospital readmission (OR 1.3, p < 0.001), and death (OR 2.1, p < 0.001) within 180 days of primary TSA. Multivariate analysis suggests these correlations are independent risk factors and not due to patient demographics or preoperative medical comorbidities. While discharge with HHC was found to be associated with increased hospital LOS (1.8 vs. 1.3 days, p < 0.001), there were no significant observed differences in cost of care. CONCLUSION: This study demonstrates that discharge with HHC compared to routine discharge while accounting for several preoperative comorbidities and demographic variables is associated with increased medical complications, SSI, readmission, and death within 180 days of TSA, but no increase in overall patient cost. These findings suggest HHC disposition status can serve as a prognosticator for postoperative complications and can help guide clinician decision making when determining appropriate surgical candidacy.

Single session of direction-specific training using auditory cues improves anticipatory postural adjustments to lateral perturbations.

When exposed to a predictable external perturbation, humans typically generate anticipatory postural adjustments (APAs) to minimize potential body disturbance. After a single session of training, individuals demonstrated the ability to rely solely on an auditory cue to elicit appropriate APAs in response to an external postural perturbation. However, whether the generation of APAs requires directional specific training remains unclear. The aim of this study was to assess whether directional-specific training with auditory cues is necessary for the generation of appropriate APA responses. Ten young adults were exposed to external perturbations targeting either their left or right shoulders, with or without an auditory cue prior to the physical impact. Electromyography (EMG) activities of sixteen trunk and leg muscles and center-of-pressure (COP) displacements were recorded and analyzed during the anticipatory and compensatory phases of postural control. Outcome measures included the latencies and integrals of muscle activities, COP displacements, and indices of co-contraction and reciprocal activation of muscles. The results revealed that, after training with right-side perturbations accompanied by an auditory cue, young adults exhibited earlier and more efficient APA responses to right-side perturbations relying only on the auditory cue. Additionally, they displayed earlier APA responses in some muscles to left-side perturbations, although these responses were less efficient. Our findings suggest that young adults could generate effective APAs to external perturbations relying on an auditory cue after a single training session; however, these responses were directional specific.

Iodine-Rich Supramolecular Complexes of Chlorobismuthates(III): Unusual Lone Pair Activity and Features of Halogen Bonding in Crystals.

The first chrorobismuthate(III) supramolecular hybrids with halogen-bond-linked I2, (Py(CH2)2Py){[BiCl5](I2)} (1) and (Py(CH2)3Py){[BiCl5](I2)0.5} (2), were synthesized via the reaction between Bi2O3, I2, and salts of the corresponding cations in HCl. The compounds featured one- and two-dimensional (2D) supramolecular motifs built by I···Cl contacts; in the case of 1, Bi(III) is pentacoordinated due to the lone pair activity, which also interacted with I2 pi-orbitals, as confirmed by density functional theory (DFT) calculations. Both complexes exhibited moderate thermal stabilities and relatively narrow optical band gaps.

Double Spin with a Twist: Synthesis and Characterization of a Neutral Mixed-Valence Organic Stable Diradical.

A convenient design strategy opens access to neutral open-shell mixed-valence species via the redox transformation of charged stable precursors, i.e., the spiro-fused borate anions. We have implemented this strategy for the synthesis of the first neutral mixed-valence diradical: two neutral mixed-valence radical fragments were assembled via a twisted biphenyl bridge. The diradical is a crystalline solid obtained in almost quantitative yield by using a facile synthetic procedure. It is stable at room temperature in the triplet ground state with a very small singlet/triplet gap. This metal-free diradical can reversibly form five redox states. The diradical exhibits an intense IVCT band in the NIR region and can be assigned as a Class 2 Robin-Day MV (mixed valence) system with weakly interacting redox centers. Computations suggest that this diradical finds itself in a unique tug-of-war between two electron delocalization patterns, Kekulé and non-Kekulé, which gives rise to two geometric isomers that are close in energy but drastically different in spin distribution and polarity. Such bistable spin-systems should be intrinsically switchable and promising for the design of functional spin devices. The scope and limitations of the new redox-strategy for the neutral MV radicals were also tested on other types of spiro-fused borates, revealing structural factors responsible for the evolution from transient to persistent and then to stable radicals.

Phosphine selenides: versatile NMR probes for analyzing hydrogen OH⋯Se and halogen I⋯Se bonds.

Nuclear magnetic resonance (NMR) spectroscopy is a powerful tool for studying the structure and dynamics of various non-covalent interactions. However, often spectral parameters that are applicable for estimation of parameters of one type of non-covalent interaction will be inapplicable for another. Therefore, researchers are compelled to use spectral parameters that are specifically tailored to the type of non-covalent interaction being studied. This complexity makes it difficult to compare different types of non-covalent interactions with each other and, consequently, to establish a strict unified classification for them. This pioneering study proposes to use phosphine selenides as universal probes for investigating hydrogen and halogen bonding in solution. The study was carried out using the example of triethylphosphine selenide Et3PSe complexes with hydrogen bonds of Se⋯HO type and R3PSe (where R: Me, Et, n-Bu, t-Bu and Ph) with halogen bonds of Se⋯X type (where X: I and Br) in solution. The presence of non-covalent interactions was confirmed experimentally by means of 1H, 31P and 77Se NMR, as well as by quantum chemical calculation methods (optimization: PW6B95-D3/def2-QZVP; NMR: B97-2/pcsSeg-2).

Synergistic cross-kingdom host cell damage between Candida albicans and Enterococcus faecalis.

The fungus Candida albicans and the Gram-positive bacterium Enterococcus faecalis share mucosal niches in the human body. As opportunistic pathogens, both are found to expand population size during dysbiosis, and can cause severe systemic infections in susceptible individuals. Here, we show that the presence of C. albicans results in increased host cell damage by E. faecalis . Furthermore, E. faecalis aggravates oropharyngeal candidiasis in mice. Increased damage is mediated by enterococcal cytolysin, and involves both physical interaction and altered glucose availability. Physical interaction promotes accumulation of bacteria on host cells, facilitating contact of cytolysin with host cells. Glucose depletion by the metabolic activity of the fungus sensitized host cells to cytolysin. This work illustrates how a complex interplay between fungi and bacteria can result in detrimental consequences for the host.

A Phenome-Wide Association Study (PheWAS) of Genetic Risk for C-Reactive Protein in Children of European Ancestry: Results From the ABCD Study.

BACKGROUND: C-reactive protein (CRP) is a moderately heritable marker of systemic inflammation that is associated with adverse physical and mental health outcomes. Identifying factors associated with genetic liability to elevated CRP in childhood may inform our understanding of variability in CRP that could be targeted to prevent and/or delay the onset of related health outcomes. METHODS: We conducted a phenome-wide association study (PheWAS) of genetic risk for elevated CRP (i.e. CRP polygenic risk score [PRS]) among children genetically similar to European ancestry reference populations (median analytic n = 5,509) from the Adolescent Brain and Cognitive Development℠ (ABCD) Study. Associations between CRP PRS and 2,377 psychosocial and neuroimaging phenotypes were estimated using independent mixed effects models. Post hoc analyses examined whether: (1) covarying for measured body mass index (BMI) or removing the shared genetic architecture between CRP and BMI altered phenotypic associations, (2) sex moderated CRP PRS associations, and (3) associations are unconfounded by assortative mating or passive gene-environment correlations (using a within-family analyses). Multiple testing was adjusted for using Bonferroni and false discovery rate (FDR) correction. RESULTS: Nine phenotypes were positively associated with CRP PRS after multiple testing correction: five weight- and eating-related phenotypes (e.g. BMI, overeating), three phenotypes related to caregiver somatic problems (e.g. caregiver somatic complaints), as well as weekday video watching (all ps = 1.2 × 10-7 - 2.5 × 10-4, all p FDR s = 0.0002 - 0.05). No neuroimaging phenotypes were associated with CRP PRS (all ps = 0.0003 - 0.998; all p FDR s = 0.08 - 0.998) after correction for multiple testing. Eating and weight-related phenotypes remained associated with CRP PRS in within-family analyses. Covarying for BMI resulted in largely consistent results, and sex did not moderate any CRP PRS associations. Removing the shared genetic variance between CRP and BMI attenuated all relationships; associations with weekday video watching, caregiver somatic problems and caregiver report that the child is overweight remained significant while associations with waist circumference, weight, and caregiver report that child overeats did not. DISCUSSION: Genetic liability to elevated CRP is associated with higher weight, eating, and weekday video watching during childhood as well as caregiver somatic problems. These associations were consistent with direct genetic effects (i.e., not solely due to confounding factors like passive gene-environment correlations) and were independent of measured BMI. The majority of associations with weight and eating phenotypes were attributable to shared genetic architecture between BMI and inflammation. The relationship between genetics and heightened inflammation in later life may be partially attributable to modifiable behaviors (e.g. weight and activity levels) that are expressed as early as childhood.

Engineering potent chimeric antigen receptor T cells by programming signaling during T-cell activation.

Programming cell signaling during T-cell activation represents a simple strategy for improving the potency of therapeutic T-cell products. Stim-R technology (Lyell Immunopharma) is a customizable, degradable synthetic cell biomimetic that emulates physiologic, cell-like presentation of signal molecules to control T-cell activation. A breadth of Stim-R formulations with different anti-CD3/anti-CD28 (αCD3/αCD28) antibody densities and stoichiometries were screened for their effects on multiple metrics of T-cell function. We identified an optimized formulation that produced receptor tyrosine kinase-like orphan receptor 1 (ROR1)-targeted chimeric antigen receptor (CAR) T cells with enhanced persistence and polyfunctionality in vitro, as assessed in repeat-stimulation assays, compared with a benchmark product generated using a conventional T-cell-activating reagent. In transcriptomic analyses, CAR T cells activated with Stim-R technology showed downregulation of exhaustion-associated gene sets and retained a unique subset of stem-like cells with effector-associated gene signatures following repeated exposure to tumor cells. Compared with the benchmark product, CAR T cells activated using the optimized Stim-R technology formulation exhibited higher peak expansion, prolonged persistence, and improved tumor control in a solid tumor xenograft model. Enhancing T-cell products with Stim-R technology during T-cell activation may help improve therapeutic efficacy against solid tumors.

A Multi-Omics Study of Epigenetic Changes in Type II Alveolar Cells of A/J Mice Exposed to Environmental Tobacco Smoke.

Lung cancer remains a major contributor to cancer fatalities, with cigarette smoking known to be responsible for up to 80% of cases. Based on the ability of cigarette smoke to induce inflammation in the lungs and increased lung cancer incidence in smokers with inflammatory conditions such as COPD, we hypothesized that inflammation plays an important role in the carcinogenicity of cigarette smoke. To test this hypothesis, we performed multi-omic analyses of Type II pneumocytes of A/J mice exposed to cigarette smoke for various time periods. We found that cigarette smoke exposure resulted in significant changes in DNA methylation and hydroxymethylation, gene expression patterns, and protein abundance that were partially reversible and contributed to an inflammatory and potentially oncogenic phenotype.

Modulation of blood-tumor barrier transcriptional programs improves intra-tumoral drug delivery and potentiates chemotherapy in GBM.

Glioblastoma (GBM) is the most common malignant primary brain tumor. GBM has an extremely poor prognosis and new treatments are badly needed. Efficient drug delivery to GBM is a major obstacle as the blood-brain barrier (BBB) prevents passage of the majority of cancer drugs into the brain. It is also recognized that the blood-brain tumor barrier (BTB) in the growing tumor represents a challenge. The BTB is heterogeneous and poorly characterized, but similar to the BBB it can prevent therapeutics from reaching effective intra-tumoral doses, dramatically hindering their potential. Here, we identified a 12-gene signature associated with the BTB, with functions related to vasculature development, morphogenesis and cell migration. We identified CDH5 as a core molecule in this set and confirmed its over-expression in GBM vasculature using spatial transcriptomics of GBM patient specimens. We found that the indirubin-derivative, 6-bromoindirubin acetoxime (BIA), could downregulate CDH5 and other BTB signature genes, causing endothelial barrier disruption in endothelial monolayers and BBB 3D spheroids in vitro. Treatment of tumor-bearing mice with BIA enabled increased intra-tumoral accumulation of the BBB non-penetrant chemotherapeutic drug cisplatin and potentiated cisplatin-mediated DNA damage by targeting DNA repair pathways. Finally, using an injectable BIA nanoparticle formulation, PPRX-1701, we significantly improved the efficacy of cisplatin in patient-derived GBM xenograms and prolonged their survival. Overall, our work reveals potential targets at the BTB for improved chemotherapy delivery and the bifunctional properties of BIA as a BTB modulator and potentiator of chemotherapy, supporting its further development.

Accounting for extinction dynamics unifies the geological and biological histories of Indo-Australian Archipelago.

Biogeographical reconstructions of the Indo-Australian Archipelago (IAA) have suggested a recent spread across the Sunda and Sahul shelves of lineages with diverse origins, which appears to be congruent with a geological history of recent tectonic uplift in the region. However, this scenario is challenged by new geological evidence suggesting that the Sunda shelf was never submerged prior to the Pliocene, casting doubt on the interpretation of recent uplift and the correspondence of evidence from biogeography and geology. A mismatch between geological and biogeographical data may occur if analyses ignore the dynamics of extinct lineages, because this may add uncertainty to the timing and origin of clades in biogeographical reconstructions. We revisit the historical biogeography of multiple IAA taxa and explicitly allow for the possibility of lineage extinction. In contrast to models assuming zero extinction, we find that all of these clades, including plants, invertebrates and vertebrates, have a common and widespread geographic origin, and each has spread and colonized the region much earlier than previously thought. The results for the eight clades re-examined in this article suggest that they diversified and spread during the early Eocene, which helps to unify the geological and biological histories of IAA.

Rhodium-Catalyzed Double Dearomatization of 1,2,3-Triazole-Isoxazole Dyads: Synthesis of Nonfused 1H-1,3-Diazepines.

A double dearomatization of dyads consisting of 1-sulfonyl-1,2,3-triazoles and 3-aryl-5-methoxyisoxazoles was applied for the efficient synthesis of nonfused 1H-1,3-diazepines. The plausible mechanism of the cascade reaction includes transformation of the 1,2,3-triazole to rhodium azavinyl carbene, the Z-selective hydride shift to form the 1-azabuta-1,3-diene moiety, rhodium-catalyzed ring contraction of the isoxazole to azirine, and pseudopericyclic four-atom ring expansion of the azirine. The synthetic utility and antiproliferative activity of the 1,3-diazepines obtained have been demonstrated.

Factors Defining Density of Retinal Macrophage-like Cells Displayed with Optical Coherence Tomography.

PURPOSE: To study the factors which define the density of MLC of the inner retinal surface in healthy eyes. METHODS: Healthy individuals, including candidates for LASIK surgery, and post-LASIK patients were included. MLC density was calculated using structural en face projections of OCT angiography scans. The status of the vitreoretinal interface was assessed as the distance from the inner limiting membrane to the posterior hyaloid membrane on cross-sectional scans and as the area of tight posterior vitreous adhesion on en face projections. The correlation between MLC density and various demographic and anatomical parameters, including the status of the vitreoretinal interface was calculated. RESULTS: Fifty-four healthy individuals, 30 post-LASIK patients all without posterior vitreous detachment (PVD) as well as 20 patients with partial PVD were included. MLC density showed a statistically significant correlation with axial length, refractive error, age, subfoveal choroidal thickness, and the status of the vitreoretinal interface (p<0.05) in eyes without PVD. In multiple regression analysis the axial length was the main parameter independently correlated with MLC density (p=0.025). The status of the vitreoretinal interface had a statistically significant correlation with the axial length (p<0.001). Partial PVD was associated with almost complete loss of MLC (p<0.001). CONCLUSION: The status of the vitreoretinal interface is a characteristic directly defining the density of retinal MLC in healthy eyes. However, axial length appears to be a key anatomical parameter which correlates with MLC density due to its effects on the adhesion of the posterior hyaloid membrane to the retinal surface.

Control Morphology and Biological Properties of HKUST-1 MOFs Using an Ultrasound-Assisted Approach.

The synthesis of bioinspired metal-organic frameworks (MOFs) performed in mild conditions with a high quality is greatly demanded. Moreover, the influence of the morphology and structure of bio-MOFs on the cell interaction and toxicity is important to determine. In this work, we developed an ultrasound (US)-assisted synthesis of HKUST-1 MOFs under mild conditions and investigated the influence of the parameters of synthesis on the morphology, structure, and biological properties of the developed MOFs. It was found that the US power, reaction time, temperature, and type of solvent composition would affect the morphology, size, and yield of the obtained crystals. Employing the optimal synthetic conditions, five types of HKUST-1 MOFs were prepared, achieving highest yields (67.8-96.2%) and different morphologies (octahedral, dodecahedral, icosahedral). The relationship between the morphological features and biological properties of developed bio-MOFs was evaluated and discussed. The cellular association and cytotoxicity of MOF@US and MOF@US-PARG were studied on various cell cultures, i.e. normal mouse embryonic fibroblasts (MEF NF2), chronic myeloid leukemia (K562), and mouse melanoma (B16-F10). The experimental results showed that MOF@US-PARG has a higher percentage of association compared to MOF@US. It has also been shown that the cytotoxicity depends on the concentration and surface modification of the developed MOFs.

Long Term Outcomes of Magnetic Sphincter Augmentation: A comparative study to Nissen Fundoplication.

BACKGROUND: Magnetic sphincter augmentation (MSA) demonstrates improvement in GERD across multiple short-term studies. Long-term, single arm studies show durable outcomes, but there is limited comparative data to Nissen (NF). METHODS: We performed a retrospective propensity matched cohort study of patients with GERD undergoing MSA or NF between 2012 and 2018. Patients were matched on age, gender, BMI, size of hiatal hernia, length of Barrett's and motility in a 1-to-1 fashion. A total of 523 patients (177 MSA, 346 NF) underwent surgery and after matching 177 MSA and 177 NF were analyzed. RESULTS: At 1 year, GERD quality of life scores improved (22 to 5 MSA vs 24 to 5 NF, p=0.593). PPI use was 14% vs 5% (p=0.010). pH testing demonstrated improved DeMeester scores (42 to 21 vs 46 to 7, p<0.001). At 5 years, GERD quality of life scores were stable (5 to 5 vs 5 to 4, p=0.208). PPI use was 31% vs 26% (p=0.474). The incidence of endoscopic dilation was similar between MSA and NF (7% vs 10%, p=0.347). Reoperation rates were higher for MSA (10% vs 4%, p=0.022) and recurrent hiatal hernias were found in 18% vs 7% (p=0.007). Compared to NF, MSA undergoing complete dissection showed no difference in dilation (5% MSA vs 7% NF, p=0.527), reoperation (8% MSA vs 6% NF, p=0.684) or hernia recurrence (10% MSA vs 6% NF, p=0.432). CONCLUSIONS: MSA achieves similar improvements in quality of life and freedom from medical therapy compared to NF especially with complete hiatal repair.

Early Optical Coherence Tomography Signs of Erdafitinib-Induced Retinopathy.

A 64-year-old male presented for a baseline ophthalmic exam before beginning erdafitinib, a fibroblast growth factor receptor (FGFR) inhibitor, for stage 4 papillary urothelial cancer. Baseline optical coherence tomography (OCT) and ophthalmic examination were unremarkable. After one month of treatment, his OCT demonstrated a significant thickening of the ellipsoid zone and prominence of the interdigitation zones along with a small amount of subretinal fluid. Two months after discontinuation of the medication, the OCT returned to baseline. Erdafitinib is a Food and Drug Administration (FDA)-approved treatment for unresectable or metastatic urothelial cancer with FGFR2 or FGFR3 mutations. However, retinal toxicity can ensue with the initiation of the drug and cause subjective vision changes and OCT abnormalities. The drug may exert toxic effects on retinal pigment epithelium, which may be seen through interval OCTs and visualization of the interdigitation zone. Therefore, pronunciation of the ellipsoid and interdigitation zone on OCT may allow for surveillance of early manifestations of erdafitinib-induced retinal toxicity.

Heterocycle-guided synthesis of m-hetarylanilines via three-component benzannulation.

A one-pot three-component synthesis of substituted meta-hetarylanilines from heterocycle-substituted 1,3-diketones has been developed. The electron-withdrawing power of the heterocyclic substituent (which can be estimated on the basis of calculated Hammett constants) in the 1,3-diketone plays a pivotal role in the studied reaction. The series of meta-hetarylanilines prepared (21-85% isolated yield) demonstrates the synthetic utility of the developed method.

An influence of electronic structure theory method, thermodynamic and implicit solvation corrections on the organic carbonates conformational and binding energies.

An impact of an electronic structure or force field method, gas-phase thermodynamic correction, and continuum solvation model on organic carbonate clusters (S)n conformational and binding energies is explored. None of the tested force field (GFN-FF, GAFF, MMFF94) and standard semiempirical methods (PM3, AM1, RM1, PM6, PM6-D3, PM6-D3H4, PM7) can reproduce reference RI-SCS-MP2 conformational energies. Tight-binding GFNn-xTB methods provide more realistic conformational energies which are accurate enough to discard the least stable conformers. The effect of thermodynamic correction is moderate and can be ignored if the gas phase conformational stability ranking is a goal. The influence of continuum solvation is stronger, especially if reinforced with the Gibbs free energy thermodynamic correction, and results in the reduced spread of conformational energies. The cluster formation binding energies strongly depend on a particular approach to vibrational thermochemistry with the difference between traditional harmonic and modified scaled rigid - harmonic oscillator approximations reaching 10 kcal mol-1.