Click synthesis, antimicrobial, DNA photocleavage and computational studies of oxindole-tethered 1H-1,2,3-triazoles

Structural figures and data set of NMR studies</p

Evaluation of cellular stress responses in magnetomotive ultrasound Supplementary figure

Early and accurate diagnoses are important for successful cancer treatment. Lymph node involvement is often critical, and magnetomotive ultrasound (MMUS) has been proposed for its detection and characterization. MMUS relies on a magnetic contrast agent, for example, iron oxide nanoparticles, delivered to the tissue of interest, magnetically set in motion and detected using ultrasound. The magneto–mechanical interaction has not previously been evaluated on a cellular level. Here we demonstrate uptake and dose-dependent retention of magnetic nanoparticles in two human adenocarcinoma cell lines, with Further, the oxidative stress levels were not affected by magnetic particles or force. Thus, we found no evidence of adverse effects from the magneto–mechanical interactions under these conditions.</p

Long-term effects of COVID-19 on lungs and the clinical relevance: a 6-month prospective cohort study - Supplemental Figure 1

Background: We aimed to explore the prevalence of prolonged symptoms, pulmonary impairments and residual disease on chest tomography (CT) in COVID-19 patients at 6 months after acute illness. Methods: In this prospective, single-center study, hospitalized patients with radiologically and laboratory-confirmed COVID-19 were included. Results: A high proportion of the 116 patients reported persistent symptoms (n = 54; 46.6%). On follow-up CT, 33 patients (28.4%) demonstrated residual disease. Multivariate analyses revealed that only neutrophil-to-lymphocyte ratio was an independent predictor for residual disease. Conclusion: Hospitalized patients with mild/moderate COVID-19 still had persistent symptoms and were prone to develop long-term pulmonary sequelae on chest CT. However, it did not have a significant effect on long-term pulmonary functions.</p

Supplementary Figures and Tables: Isothermal amplification using sequence-specific fluorescence detection of SARS-CoV-2 and variants in nasal swabs

      Supplemental   Figure 1. FQ-LAMP LOD for WA1 - Reaction Tube Images WA1 Target Log Dilution in SPS. Culture   supernatants were log-serially diluted from 106 /ml to 10 pfu/ml   in the SPS buffer and analyzed by FQ-LAMP. B. WA1   Target Two-fold Dilution in SPS.  WA1-infected Vero cell culture supernatant two-fold   diluted in SPS buffer from 1,000 to 32 PFU/ml was processed with the SPS   protocol and analyzed by FQ-LAMP C. No   Template Control. FQ-LAMP assay   using identically processed samples, but lacking WA1 with deionized water   added to the SPS buffer served as the no-template control (NTC). Images were acquired using a mobile phone   camera with no filters and excitation using a handheld UV lamp (302nm). Supplemental Figure 2. FQ-LAMP   WA1 Target Multiples to Confirm LOD – Reaction Tube Images.  FQ-LAMP assay targeting WA1 from infected Vero   cells. Culture supernatants were spiked into pooled CoV2-negative nasal swab   samples at: A. 2,000; B. 1,000;   and C. 500 PFU/ml representing 2X,   1X, and 0.5X LOD target concentration, respectively. All samples were   processed with SPS buffer as described above using n=20 replicates for each   target dilution. D. Negative control assays (no spike), n=20   replicates used identically processed nasla swab sample but without WA1. Images acquired using a mobile phone camera   with no optical filters under excitation with a laboratory handheld UV lamp   (302nm). 1,000 PFU/ml WA1 target   concentration was the highest dilution displaying at least 19/20 replicates   with positive signal and was determined to be the assay LOD.  Supplemental Figure 3. FQ-LAMP   CoV2 Variants in Nasal Swab Sample (2,000 PFU/ml) Reaction Tube Images FQ-LAMP targeting CoV2   variant viruses:  A. Delta (B.1.617.2), B.  Alpha (B.1.1.7), and C. Omicron (B.1.1.529) from infected Vero cell cultures. Culture supernatants were spiked into   pooled CoV2-negative nasal swab sample at 2,000 PFU/ml (2X LOD for WA1) and   processed with SPS buffer as described above using n=20 replicates for each   target virus. Images acquired using a   mobile phone camera with no filters under excitation with a standard   laboratory handheld UV lamp (302nm). CoV2 variant viruses are easily detectable by FQ-LAMP in nasal swab   sample background at 2X LOD, and all the heterologous virus targets, even at   very high concentrations, failed to produce fluorescence signals over the   background in the FQ-LAMP assay (images not shown).  Supplemental Table 1. FQ-LAMP targeting WA1 from infected Vero cells.   Culture supernatants were spiked into pooled CoV2-negative nasal swab samples   at 2,000, 1,000; and 500 PFU/ml representing 2X, 1X, and 0.5X LOD target   concentration, respectively, and processed with SPS buffer as described using   n=20 replicates for each target dilution (three representative samples are   pictured).  1,000 PFU/ml WA1 target   concentration was the highest dilution displaying at least 19/20 replicates   with a positive signal in the LOD. FQ-LAMP assay targeting CoV2 variant viruses Delta (B.1.617.2), Alpha   (B.1.1.7), and Omicron (B.1.1.529) from infected Vero cell culture. Culture supernatants were spiked into   pooled CoV2-negative nasal swab sample at 2,000 PFU/ml (2X LOD for WA1) and   processed with SPS buffer as described above using n=20 replicates for each   target virus, and FQ-LAMP targeting three human coronavirus strains i.e. OC43   (GenBank: AY585228.1), 229E (GenBank: AF304460.1), and NL63 (GenBank:   AY567487.2) from infected Vero cells, human respiratory syncytial virus   strain A2, GenBank: KT992094.1) from infected Vero cells, and influenza A   virus A/Guangdong-Maonan/SWL1536/2019 (H1N1, GISAID: EPI_ISL_419003) from   infected embryonated hen eggs. Culture supernatants, or egg lysate for IAV,   were spiked into pooled SARS-CoV-2 negative nasal swab sample at 105 PFU/ml   and processed with SPS buffer as described using n=20 replicates for each   target virus, and negative control assays (no spike), n=20 replicates   consisting of identically processed nasal swab sample, but without CoV2   variant virus. CoV2 variant viruses are easily detectable by FQ-LAMP in nasal   swab sample background at 2X LOD, and all the heterologous virus targets,   even at very high concentrations, did not produce a fluorescence signal over the background in   the FQ-LAMP assay.   Supplemental Table 2. CoV2 N   gene-based FQ-LAMP primers and signal oligos were created using free Primer   Explorer V software. Numbering   according to CoV2 Wuhan-Hu-1 (GenBank: MN908947.3). </p

A Review on Non-Viral, Non-Bacterial Infectious Agents Toxicity Involved in Neurodegenerative Diseases - supplementary material

Supplementary table </p

Supplementary Materials: Validation of CRISPR targeting for proliferation and cytarabine resistance control genes in the AML cell line MOLM-13

Supplementary figure 1. Control genes’ mRNA expression relative to HPRT in MOLM-13 cells. GAPDH was used as reference gene for data normalization. Data represents mean ± SEM (n = 3). Supplementary figure 2. p53 depletion induces MOLM-13 cells’ proliferation. (A) Density plot of EdU incorporation analysis of p53 (p53-1, -2 and -3) depleted MOLM-13 cells. LentiCRISPRv2-GFP (EP) and three independent NS sgRNA lentiCRISPRv2-GFP (NS1-3) constructs transduced cells served as controls. EdU untreated MOLM-13 cell sample was used as control to gate EdU positive cells. (B) Bar graph shows relative quantitation of EdU incorporation. Data represents mean ± SEM (n = 6) and are from two independent triplicate experiments. (C) Colorimetric proliferation analysis of p53 depleted cells compared to control cells. EP and three independent NS sgRNA constructs transduced cells served as controls. Data represents mean ± SEM (n = 6) and are from two independent triplicate experiments. (D) Flow cytometric cell count analysis of control versus p53 depleted MOLM-13 cells. EP and NS sgRNA constructs transduced cells served as controls. Data represents mean ± SEM (n = 6) and are from two independent triplicate experiments. Significance testing was performed using student's t-test **p<0.01, ***p<0.001. EP = empty plasmid; NS= non-specific; EP = empty plasmid. Supplementary figure 3. DCK depletion induces MOLM-13 cells’ resistance to Ara-C. (A) Ara-C kill curve of MOLM-13 cells. Dashed lines’ intersection indicates half maximal inhibitory concentration (IC50) of Ara-C for MOLM-13 cells. (B) Colorimetric survival analysis of DCK depleted MOLM-13 cells treated with Ara-C. lentiCRISPRv2-GFP (EP) and NS sgRNA lentiCRISPRv2-GFP constructs transduced cells served as controls. Data represents mean ± SEM (n = 6) and are from two independent triplicate experiments. (C) Flow cytometric cell count analysis of control (NS) versus DCK depleted MOLM-13 cells subjected to Ara-C treatment. EP and NS sgRNA constructs transduced cells served as controls. Data represents mean ± SEM (n = 6) and are from two independent triplicate experiments. ***p<0.001 Ara-C treated DCK group was compared with Ara-C treated control group. EP = empty plasmid; NS= non-specific sgRNAs. Supplementary figure 4. MS2 expression in lentiMPHv2 stably transduced MOLM-13 cells. Shown is relative mRNA transcript level of MS2 relative to HPRT in a representative MOLM-13 sample transduced with lentiMPHv2. GAPDH was used as the reference gene for data normalization. Data represents mean ± SEM (n = 3). Supplementary figure 5. CDA upregulation induces MOLM-13 cells’ resistance to Ara-C. (A) CDA mRNA expression (q-PCR) in activation constructs transduced lentiMPHv2 MOLM-13 cells. LentiSAMv2 (EP) and three independent NSa sgRNA LentiSAMv2 constructs transduced cells served as controls. Data represents mean ± SEM (n = 6) and are from two independent triplicate experiments. (B) Colorimetric survival analysis of activation constructs transduced lentiMPHv2 MOLM-13 cells treated with Ara-C. EP and three NSa sgRNA constructs transduced cells served as controls. Data represents mean ± SEM (n = 6) and are from two independent triplicate experiments. (C) Flow cytometric cell count analysis of control versus CDA upregulated MOLM-13 cells subjected to Ara-C treatment. EP and NSa sgRNA constructs transduced cells served as controls. Data represents mean ± SEM (n = 6) and are from two independent triplicate experiments. ***p<0.0001 Ara-C treated CDA group was compared with Ara-C treated control group. Comparison testing was performed using student's t-test. EP = empty plasmid; NSa = non-specific sgRNAs. Supplementary Table 1. Supplementary Table 1. Results from experimental comparison testing. Statistics of experimental results and results from paired T-tests performed. Shown are experimental and control group means and standard deviation as well as results from comparative testing performed (t statistic and p-value from paired t-tests). Supplementary Table 2. Reagents, supplies and equipment table. Information on reagents, supplies and equipment used in described protocol with annotation to respective protocol step. Protocol for Validation of CRISPR targeting for proliferation and cytarabine resistance control genes in the AML cell line MOLM-13. </div

Design and generation of mRNAs encoding conserved regions of SARS-CoV-2 ORF1ab for T cell-mediated immune activation: supplementary materials

Aim: To generate mRNAs encoding conserved regions within SARS-CoV-2 ORF1ab which can induce strong T-cell responses to overcome the immune invasion of newly emergent variants. Methods:We selected two conserved regions with a high density of T-cell epitopes using immunoinformatics for mRNA synthesis. The ability of testing mRNAs to activate T cells for IFN-γ production was examined by an ELISpot assay and flow cytometry. Results: Two synthesized mRNAs were successfully translated in MDA-MB-231 cells and had comparable potency to the spike mRNA to induce CD4+ and CD8+ T-cell responses in PBMCs in 29 out of 34 participants. Conclusion: This study provides a proof-of-concept for the use of SARS-CoV-2 conserved regions to develop booster vaccines capable of eliciting T-cell-mediated immunity.</p

Fabrication, characterization and wound-healing properties of core–shell SF@chitosan/ZnO/Astragalus arbusculinus gum nanofibers

Percutaneous coronary intervention with implantation of second-generation drug-eluting stents (DES) has emerged as a mainstay for the treatment of obstructive coronary artery disease given its beneficial impact on clinical outcomes in these patients. Everolimus-eluting stents (EES) are one of the most frequently implanted second-generation DES; their use for the treatment of a wide range of patients including those with complex coronary lesions is supported by compelling evidence. Although newer stent platforms such as biodegradable polymer DES may lower local vessel inflammation, their efficacy and safety have not yet surpassed that of Xience stents. This article summarizes the properties of the Xience family of EES and the evidence supporting their use across diverse patient demographics and coronary lesion morphologies. Plain language summary: Patients with coronary artery disease (CAD) often require treatment for symptoms caused by blockages in coronary arteries. In addition to medical therapy, available procedure options include either coronary artery bypass grafting, a major heart surgery or percutaneous coronary intervention (PCI) with stenting. PCI is a minimally invasive procedure where ametallic stent (ameshmade up of fine metallic network in a tube shape used to keep vessels open) is advanced over a wire through an artery to open the coronary artery blockage. Over the past few decades, improvements in procedure technique and stent material have made PCI a highly safe and efficacious procedure. A newer generation of stents, known as drug-eluting stents (DES), have been developed in which metallic struts are covered with a highly biocompatible polymer (a thin material coating over the metallic mesh) that releases drugs at the blockage site to prevent local cell growth in the vessel wall. Among the second-generation DES, Xience everolimus-eluting stents (EES) have shown better outcomes compared with earlier generations of stents. Another version of DES with biodegradable polymer coating is emerging but their advantage over EES remains uncertain. Currently, Xience EES are one of the most commonly used stents to treat CAD. This manuscript covers an in-depth review of clinical evidence on the performance of Xience stents in a diverse range patient populations.</p

Efficacy and safety of mavacamten intreatment of hypertrophic cardiomyopathy:a systematic review and meta-analysis: supplementary data 1

Aim: This meta-analysis was performed to assess the efficacy and safety of mavacamten in patients withhypertrophic cardiomyopathy. Methods & materials: A search was conducted using PubMed, Cochrane,and Scopus up to August 2022 for randomized studies reporting our pre-specified outcomes. Results: It wasobserved that mavacamten significantly improved New York Heart Association class (p Summary Score of the Kansas City Cardiomyopathy Questionnaire (p = 0.02), post-exercise left ventricularoutflow tract gradient (p therapy rates (p events, ≥1 treatment-emergent adverse events, left ventricular volume index, left ventricular fillingpressure, left ventricular end-diastolic volume index, and peak oxygen uptake (pVO2). Conclusion: Futurelarge-scale trials are required to confirm our results and determine the long-term benefits and risks ofmavacamten use in these patients.</p

Supplementary Materials: Barriers and facilitators to the uptake of the Concussion Awareness Training Tool as continuing medical education in primary care.docx

   Supplementary Material: Physician Survey</p