Single-cell RNA-sequencing resolves self-antigen expression during mTEC development

The crucial capability of T cells for discrimination between self and non-self peptides is based on negative selection of developing thymocytes by medullary thymic epithelial cells (mTECs). The mTECs purge autoreactive T cells by expression of cell-type specific genes referred to as tissue-restricted antigens (TRAs). Although the autoimmune regulator (AIRE) protein is known to promote the expression of a subset of TRAs, its mechanism of action is still not fully understood. The expression of TRAs that are not under the control of AIRE also needs further characterization. Furthermore, expression patterns of TRA genes have been suggested to change over the course of mTEC development. Herein we have used single-cell RNA-sequencing to resolve patterns of TRA expression during mTEC development. Our data indicated that mTEC development consists of three distinct stages, correlating with previously described jTEC, mTEChi and mTEClo phenotypes. For each subpopulation, we have identified marker genes useful in future studies. Aire-induced TRAs were switched on during jTEC-mTEC transition and were expressed in genomic clusters, while otherwise the subsets expressed largely overlapping sets of TRAs. Moreover, population-level analysis of TRA expression frequencies suggested that such differences might not be necessary to achieve efficient thymocyte selection.RM is supported by a PhD Fellowship from the Fundação para a Ciência e Tecnologia, Portugal (SFRH/ BD/51950/2012). XZ is supported by an Advanced Postdoc Mobility Fellowship from the Swiss National Science Foundation (SNSF, grant number P300P2_151352). Part of the work was performed during XZ’s visit to the Simons Institute for the Theory of Computing. TL is supported by the Academy of Finland (Decision 311081). The authors would like to thank Bee Ling Ng and the staff of the Cytometry Core Facility, and Stephan Lorenz and the staff of the Single Cell Genomics Core Facility for their contribution. Mark Lynch is acknowledged for technical assistance with the Fluidigm C1 platform. Mike Stubbington and Kylie James are acknowledged for revising the language of the manuscript. We thank Sarah Teichmann for help and discussions regarding the manuscript.info:eu-repo/semantics/publishedVersio

Synthesis, Crystal structure, DFT calculations and antimicrobial activity of 4-(4-fluoro-phenyl)-2,6-dimethyl-1,4-dihydro-pyridine-3,5-dicarboxylic acid diethyl ester

The title compound was synthesized and confirmed by FT-IR, 1H, 13C NMR analysis. The molecular structure of the compound was precisely determined by Single Crystal X-ray Diffraction (SC-XRD) analysis. The crystalized compound shows P21/C & monoclinic crystal system with cell parameters a = 9.7768 (5), b = 7.4005(3) and c = 24.8099 (12), β=93.734(2)°.The structural and electronic properties of the compound were carried out by Density Functional Theory (DFT) calculations. The compound exhibited H-bonding between N1-H1A-O1 with bond distance 2.98(7) A°).The energy gap Egap 4.53eV and Egap= 4.34eV for crystal and DFT method respectively. The molecular orbitals energies were studied through Highest Unoccupied Molecular Orbital (HOMO) and Lowest Unoccupied Molecular Orbital (LUMO) analysis. The softness and hardness of the molecule was studied through Global Chemical Reactivity Descriptors (GCRD). The electrophilic and nucleophilic characters were studied through Molecular Electrostatic Potential (MEP) studies. The antimicrobial studies were carried out by in-vitro method against 6 microorganisms

The impact of direct‐acting antiviral agents on liver and kidney transplant costs and outcomes

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/146297/1/ajt14895_am.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/146297/2/ajt14895.pd

Management and outcomes of unilateral group d tumors in retinoblastoma

Purpose: Retinoblastoma presents most commonly as advanced unilateral disease, particularly in developing countries for which primary enucleation has been the preferred method of treatment. However, with the evolution of newer treatment modalities including intravitreal chemotherapy, intra-arterial chemotherapy and newer chemotherapeutic combinations, a trend towards more conservative approaches is being observed. Our aim is to evaluate outcomes of group D eyes following conservative and non-conservative treatment options. Patients and Methods: The ocular oncology database was used to identify eyes with unilateral retinoblastoma that fulfilled the International Intraocular Retinoblastoma Classification (IIRC) group D criteria from August 2010 to August 2018 and these were retrospectively reviewed. Overall, 39 eyes were identified. Results: Nineteen (49%) eyes underwent primary enucleation and 20 (51%) received eyeconserving treatment. Eye salvage was possible in 15 (75%) eyes in the attempted salvage group. None of the patient revealed signs of metastasis. All eyes received conventional chemotherapy (carboplatin, vincristine, etoposide) and focal laser therapy. Additional treatment modalities offered included intravitreal chemotherapy, intra-arterial chemotherapy and topotecan. Three (11%) eyes in the primary enucleation group showed high-risk features on histopathology and none developed metastasis. Conclusion: The results of the study seem promising and conservative measures can be adopted in selected unilateral group D eyes

Delayed intravitreal anti-vegf therapy for patients during the covid-19 lockdown: An ethical endeavor

Purpose: To assess the impact of Jordanian’s Corona Virus Disease (COVID-19) lockdown on visual acuity and macular thickness in patients with macular edema receiving intravitreal injections, and to assess the ethical endeavor of lockdown among serious sight threatening conditions. Patients and Methods: This retrospective observational study included patients planned for intravitreal injections who did not complete the planned course before the lockdown (ie, before 20th of March 2020). Data included demographics, indication for the intravitreal injection, corrected distance visual acuity (CDVA), and central macular thickness on Optical Coherence Tomography (OCT) before and after the lockdown. Results: One-hundred and sixty-six eyes of 125 patients were studied, 68 (54.4%) patients were males, and the mean (± standard deviation, SD) age was 64.79 (±9.41) years. Mean (±SD) duration of delay in the planned injection was 60.97 (±24.35) days. The change in visual acuity was statistically significant for patients with diabetic macular edema (p= 0.045 improvement), patients with central retinal vein thrombosis (CRVO) (p= 0.05 deterioration), and patients with age-related macular degeneration (AMD) (p= 0.005 deterioration). Of interest, delay of more than 2 months and the previous need for 3 or more injections were significant poor prognostic factors for visual outcome for patients with diabetic macular edema (p=0.027 and 0.045). Conclusion: The impact of delay in the scheduled intravitreal injections resulted in variable outcomes depending on the indication. Triaging the urgency of patients should be based on the indication to support the equity principle of bioethics, where those in need are prioritized against others, depending on potential adverse outcome

The Effects of Age and Sex of Megakaryocyte Secreted Factors on Endothelial Cell Growth and Function

With an aging population, the risk of fractures and compromised healing increases. Angiogenesis and vasculogenesis are impaired with aging. Vascularization at the callus plays a significant role in bone healing, and we have previously shown the important role ofmegakaryocytes (MKs) in regulating bone healing. Notably, MK-derived conditioned media (CM) contains factors known to promote angiogenesis. Whether differences exist with aging and/or based on sex was the primary focus of this study. Here, we examined the effects of CM secreted from MKs derived from young (3-4-month-old) and aged (22-24-month-old) male and female C57BL/6J mice on bone marrow endothelial cell (BMEC) growth and function. Specifically, BMEC proliferation, vessel-like formation, wound/transwell migration, and RNA expression were examined. Both young and aged female MK CM saw a >65% increase in BMEC proliferation(p<0.001 and p<0.05, respectively). In addition, female MK CM, regardless of age, improved all four parameters of vessel-like formation by >115% (p<0.05). Likewise, young male MK CM increased vessel-like formation in all parameters by more than 160% (p<0.001). Although aged male MK CM resulted in higher vessel-like formation parameters, including significant >150% increases in the formation of nodes and meshes, 62% fewer vessel-like structures formed compared to that observed with young male MK CM treatment (p<0.05). Additionally, aged MK CM, irrespective of sex, improved transwell migration by over 2500% (p<0.01). On the other hand, aged female and male MK CM inhibited wound closure by 46% and 17%, respectively (p<0.05). RNA analysis found MK CM yielded significantly different levels of expression in CXCR4, CXCR2, CD36, CD74, PDGFRB, and TGFBRB not only relative to controls, but also between sexes and ages. Further testing to identify the mechanisms responsible for these age-associated differences may allow for novel treatment strategies to improve MK-mediated angiogenesis, vasculogenesis, and bone healing, particularly within the aging population

District Highway Maintenance Research-On-Call (ROC) 2023-09 Task #8: Improving the Process of Removing Totem Poles

37852Roadside vegetation management can be a time-consuming and potentially dangerous task, particularly on steep embankments typical of unglaciated regions in Ohio. Furthermore, tree clearing is usually undertaken during difficult weather conditions (i.e., snowy, icy) as regulations for agencies that receive federal funding limit clearing to protect habitat for the Indiana and northern long-earned bat species. The combination of steep and unstable terrain can slow progress and expose workers to difficult working conditions with higher risk for accidents

The Potential Tripartite Connection: Alzheimer\u27s Disease, Fracture Healing, and the Gut Microbiome

Alzheimer\u27s disease (AD), fracture healing, and the gut microbiome are interconnected aspects of health that have gained significant research interest. Recent studies suggest gut dysbiosis may play a role in AD pathogenesis, potentially through the gut-brain axis, a bidirectional communication system. Moreover, the gut microbiome\u27s role in bone health could link dysbiosis and fracture risk. Furthermore, research reports have revealed that the brain communicates with bone, termed the bone-brain axis. Despite these insights, the effect of the gut microbiome on fracture healing in AD remains largely unexplored. To uncover these connections, our study uses the AD mouse models, 3xTg and 5xFAD. We conducted osteotomies on these mice and analyzed fecal samples that were collected at different timepoints. Fecal samples are being examined via qPCR and 16s RNA analysis toidentify and quantify bacterial phyla. These findings will be linked to both AD progression, gauged through behavior and histological analyses, and fracture healing, quantified using X-ray mRUST scoring, microCT, and histology. We hypothesize that the progression of AD could alter the gut microbiome, potentially affecting fracture healing. This might occur through inflammation pathways triggered by specific gut bacteria. We may identify specific gut bacteria that play critical roles in both fracture healing and AD. We anticipate finding a shift towards pro-inflammatory bacterial phyla in the context of AD progression and during the fracture healing process. If this hypothesis is validated, it could unlock new therapeutic strategies aimed at targeting the gut microbiome to improve bone health, fracture healing, and AD progression in patients

Evaluation of metals that are potentially toxic to agricultural surface soils, using statistical analysis, in northwestern Saudi Arabia

© 2015, Springer-Verlag Berlin Heidelberg. Heavy metals in agricultural soils enter the food chain when taken up by plants. The main purpose of this work is to determine metal contamination in agricultural farms in northwestern Saudi Arabia. Fifty surface soil samples were collected from agricultural areas. The study focuses on the geochemical behavior of As, Cd, Co, Cr, Cu, Hg, Pb and Zn, and determines the enrichment factor and geoaccumulation index. Multivariate statistical analysis, including principle component analysis and cluster analysis, is also applied to the acquired data. The study shows considerable variation in the concentrations of the analyzed metals in the studied soil samples. This variation in concentration is attributed to the intensity of agricultural activities and, possibly, to nearby fossil fuel combustion activities, as well as to traffic flows from highways and local roads. Multivariate analysis suggests that As, Cd, Hg and Pb are associated with anthropogenic activities, whereas Co, Cr, Cu and Zn are mainly controlled by geogenic activities. Hg and Pb show the maximum concentration in the analyzed samples as compared to the background concentration