Spatial imaging of glycoRNA in single cells with ARPLA.

Little is known about the biological roles of glycosylated RNAs (glycoRNAs), a recently discovered class of glycosylated molecules, because of a lack of visualization methods. We report sialic acid aptamer and RNA in situ hybridization-mediated proximity ligation assay (ARPLA) to visualize glycoRNAs in single cells with high sensitivity and selectivity. The signal output of ARPLA occurs only when dual recognition of a glycan and an RNA triggers in situ ligation, followed by rolling circle amplification of a complementary DNA, which generates a fluorescent signal by binding fluorophore-labeled oligonucleotides. Using ARPLA, we detect spatial distributions of glycoRNAs on the cell surface and their colocalization with lipid rafts as well as the intracellular trafficking of glycoRNAs through SNARE protein-mediated secretory exocytosis. Studies in breast cell lines suggest that surface glycoRNA is inversely associated with tumor malignancy and metastasis. Investigation of the relationship between glycoRNAs and monocyte-endothelial cell interactions suggests that glycoRNAs may mediate cell-cell interactions during the immune response.

Crystalline Nanoparticles of Water-Soluble Covalent Basket Cages (CBCs) for Encapsulation of Anticancer Drugs.

We herein describe the preparation, assembly, recognition characteristics, and biocompatibility of novel covalent basket cage CBC-11, composed of four molecular baskets linked to four trivalent aromatic amines through amide groups. The cage is tetrahedral in shape and similar in size to small proteins (Mw =8637 g/mol) with a spacious nonpolar interior for accommodating multiple guests. While 24 carboxylates at the outer surface of CBC-11 render it soluble in aqueous phosphate buffer (PBS) at pH=7.0, the amphiphilic nature prompts its assembly into nanoparticles (d=250 nm, DLS). Cryo-TEM examination of nanoparticles revealed their crystalline nature with wafer-like shapes and hexagonally arranged cages. Nanoparticulate CBC-11 traps anticancer drugs irinotecan and doxorubicin, with each cage binding up to four drug molecules in a non-cooperative manner. The inclusion complexation resulted in nanoparticles growing in size and precipitating. In media containing mammalian cells (HCT 116, human colon carcinoma), the IC50 value of CBC-11 was above 100 µM. While this work presents the first example of a large covalent organic cage operating in water at the physiological pH and forming crystalline nanoparticles, it also demonstrates its biocompatibility and potential to act as a polyvalent binder of drugs for their sequestration or delivery.

Dissipative Formation of Covalent Basket Cages.

Living systems use chemical fuels to transiently assemble functional structures. As a step toward constructing abiotic mimics of such structures, we herein describe dissipative formation of covalent basket cage CBC 5 by reversible imine condensation of cup-shaped aldehyde 2 (i.e., basket) with trivalent aromatic amine 4. This nanosized [4+4] cage (V=5 nm3 , Mw =6150 Da) has shape of a truncated tetrahedron with four baskets at its vertices and four aromatic amines forming the faces. Importantly, tris-aldehyde basket 2 and aliphatic tris-amine 7 undergo condensation to give small [1+1] cage 6. The imine metathesis of 6 and aromatic tris-amine 4 into CBC 5 was optimized to bias the equilibrium favouring 6. Addition of tribromoacetic acid (TBA) as a chemical fuel perturbs this equilibrium to result in the transient formation of CBC 5, with subsequent consumption of TBA via decarboxylation driving the system back to the starting state.

Advocacy in Action: a Comprehensive Student-Led Proactive Outreach to Patients at Highest Risk.

INTRODUCTION: COVID-19 disrupted access to critical healthcare and resources for many, especially affecting patients at safety-net hospitals who rely on regular care for multiple complex conditions. Students realized they could support patients from the sidelines by helping navigate abrupt healthcare changes and proactively addressing needs at home. AIM: To comprehensively identify and meet the clinical and social needs of Atlanta, Georgia's patients at highest risk, left without their usual access to healthcare, through proactive telephonic outreach. SETTING AND PATIENTS: Medical and Physician's Assistant students from Emory and Morehouse Schools of Medicine partnered with Grady Health System, Atlanta's safety-net hospital. Artificial intelligence prioritized over 15,000 patients by risk of morbidity and mortality from COVID-19. PROGRAM DESCRIPTION: In this novel program, students performed telephonic outreach to thousands of patients at highest risk of poor outcomes from COVID-19. Students used a custom REDCap form that served as both a call script and data collection tool. It provided step-by-step guidance to (1) screen for COVID-19 and educate on prevention; (2) help patients navigate health system changes to fill gaps in care; and (3) identify and address social needs. Based on patients' responses, the form prompted tailored reminders for next steps and connections to medical and social resources. PROGRAM EVALUATION: In the program's first 16 months, students made 7,988 calls, of which 3,354 were answered. Over half (53%) of patients had at least one need requiring action: 48% health and 16% social. DISCUSSION: This proactive, novel initiative identified substantial clinical and social need among patients at highest risk for poor outcomes and filled a pressing health system gap exacerbated by COVID-19. Simultaneously, interprofessional students gained applied exposure to health systems sciences. This program can serve as a model for rapid, cost-effective, high-yield outreach to promote patient health at home both during and beyond the pandemic.

Remote Learning of COVID-19 Kinetic Analysis in a Physical Chemistry Laboratory Class.

The COVID-19 pandemic has affected many in-person laboratory courses across the world. The viral spreading model is complicated but parameters, such as its reproduction number, R t, can be estimated with the susceptible, infectious, or recovered model. COVID-19 data for many states and countries are widely available online. This provides an opportunity for the students to analyze its spreading kinetics remotely. Here, we reported a laboratory set up online during the third week of the spring semester of 2021 to minimize social contacts. Due to the wide interest in developing online physical chemistry and analytical laboratories during the pandemic, we would like to share this laboratory design. The method, technique, procedure, and grading are described in this report. The student participants were able to apply the kinetic techniques learned in physical chemistry to successfully analyze an ongoing real-world problem through a remote learning environment and prepare this report.

Sustained Clinical Performance During Surgical Rotations Predicts NBME Shelf Exam Outcomes.

OBJECTIVE: To determine if longitudinal, excellent clinical performance reflected in subjective evaluations during a surgery clerkship would be associated with a greater likelihood of National Board of Medical Examiners Surgery Shelf Exam ("shelf exam") success. DESIGN: We retrospectively reviewed medical students' surgical clerkship performance from 2014 to 2019. Clinical evaluations for each rotation were abstracted and students were stratified by performance: excellent performers and non-excellent performers. The rotation performance grades were then combined to classify overall clerkship performance: sustained excellent performers, improved performers, worsened performers, and sustained non-excellent performers. We compared the shelf exam scores between performer class for each clinical rotation and the overall clerkship. Using logistic regression, we also sought to determine if clinical performance predicted passing the shelf exam. SETTING: Emory University School of Medicine in Atlanta, Georgia. PARTICIPANTS: Third-year medical students (N = 674) who completed a surgery clerkship. RESULTS: Excellent performers scored higher than non-excellent performers on the shelf exam during both clinical rotations (all p < 0.01). Sustained excellent performers had the highest exam scores out of all the clerkship performance groups (p < 0.0001). Excellent performers for both rotations were associated with increased odds of passing the shelf exam. Sustained excellent performers had the greatest odds (OR 3, 95% confidence interval 1.5-6.3, p = 0.003) of passing the exam. CONCLUSIONS: Clinical performance during the surgical clerkship and individual rotations correlates with shelf exam scores. Students should be encouraged to excel on the wards to maximize the educational experience and improve their odds of passing the exam.