Journal: Bioengineering & Translational Medicine

Article Title: Neuron‐targeted 2‐deoxyglucose‐dendrimer‐rosiglitazone nanotherapy mitigates neuroinflammation and cognitive deficits in pediatric traumatic brain injury

doi: 10.1002/btm2.70053

Figure Lengend Snippet: FIGURE 8 mRNA expressions of pro- and anti-inflammatory markers and cell death markers from sham (n = 12, 6 M/6 F), traumatic brain injury (TBI) + vehicle (n = 12, 6 M/6 F), TBI + rosiglitazone (Rosi) (n = 12, 6 M/6 F), and TBI + 2-deoxyglucose (2DG-D)-Rosi (n = 12, 6 M/6 F) groups. Neurons were isolated from the injured brain regions (or the matching area from the sham mice) at 24-h post-treatment for gene expression evaluations. Data were presented as mean ± SEM. Treatment groups were presented as sham (gray circles), TBI + vehicle (blue circles), TBI + Rosi (magenta circles), and TBI + 2DG-D-Rosi (green circles). (a–d) The expression of pro-inflammatory makers, tumor necrosis factor-alpha (TNF-α) (a), interleukin-6 beta (IL-1β) (b), Toll-like receptor 4 (TLR4) (c), and NLR Family Pyrin Domain Containing 3 (NLRP3) (d). (e) The expression of anti-inflammatory maker IL-13. (f) The expression of cell death marker Fas. *p < 0.05; **p < 0.01; ***p < 0.001. (g) The co- localization of neurodegenerative markers marker Fluoro-Jade C (FJC) (green) and neuronal marker (NeuN, red) was evaluated in sham (n = 8, 4 M/4 F), TBI + vehicle (n = 8, 4 M/4 F), TBI + Rosi (n = 8, 4 M/4 F), and TBI + 2DG-D-Rosi (n = 8, 4 M/4 F) groups at 24-h post-injury. Images (40, 5 images/animal) were randomly acquired from the cortex area (mainly primary motor cortex and primary somatosensory cortex) in the injured brain regions (approximately between bregma +1 mm and bregma 0.5 mm) using the Nikon Eclipse TS2R fluorescent microscope (Nikon, NY, USA). The representative images from male treatment groups (upper panels) and female treatment groups (lower panels). 40,6-Diamidino-2-phenylindole (DAPI) (blue) was used for nucleus stains. Scale bars: 100 μm. (h) The quantification of FJC+NeuN+ cells among treatment groups. Data were presented as mean ± SEM. Treatment groups were presented as sham (gray circles), TBI + vehicle (blue circles), TBI + Rosi (magenta circles), and TBI + 2DG-D-Rosi (green circles). *p < 0.05; **p < 0.01; ****p < 0.0001.

Article Snippet: Brain sections were washed 3 times in PBS for 5 min each, incubated in 0.001% Fluoro-Jade C (FJC) (Cat# TR-160-FJC, Biosensis, CA, USA) for 10 min, washed six times in PBS for 15 min each, and incubated with rabbit anti-NeuN (1:250; Cat# ab177487; Abcam, MA, USA).

Techniques: Isolation, Gene Expression, Expressing, Marker, Microscopy

Journal: Cell

Article Title: SARS-CoV-2 infection triggers profibrotic macrophage responses and lung fibrosis

doi: 10.1016/j.cell.2021.11.033

Figure Lengend Snippet: CD163 + macrophages accumulate in the lung in severe COVID-19 (A) Overview of study design and analyses. CT, computed tomography; BAL, bronchoalveolar lavage; scRNA-seq, single-cell RNA sequencing; snRNA-seq, single-nucleus RNA sequencing; IHC, immunohistochemistry; IF, immunofluorescence microscopy; MELC, multi-epitope ligand cartography; EM, electron microscopy; VCin, inspiratory vital capacity; PBMC, peripheral blood mononuclear cells; IAV, Influenza A virus. (B) Postmortem analysis of consecutive histological sections of non-COVID-19 (left) and COVID-19 autopsy lung samples (right) by hematoxylin and eosin (H&E; top) and CD68 IHC (bottom). Scale bar, 100 μm. (C) IF of CD68 (green) and CD163 (red) in lung tissue autopsy samples of COVID-19 patients and non-COVID-19 controls. Arrows indicate CD68 + CD163 – macrophages, and arrowheads indicate CD68 + CD163 + macrophages. Scale bar, 20 μm. (D) Quantification of CD68 + macrophage density (left) and the proportion of CD163 + macrophages (right) in lung autopsy samples from fifteen donors (as in C). Mann-Whitney test; ∗ p < 0.05. (E) Representative images of consecutive histological sections of lung autopsy samples. H&E (left), CD68 IHC (middle), and SARS-CoV-2 RNA-FISH (right). Arrowheads indicate SARS-CoV-2 RNA-positive macrophages. Scale bars, 50 μm, 25 μm. RNA-FISH, RNA-fluorescence in situ hybridization. (F) Lung autopsy samples of 9 COVID-19 patients were analyzed by MELC with a panel of 22 markers on 19 fields of view (FOVs). Two-dimensional embedding computed by UMAP on 9,684 computationally identified CD45 positive cells (T cells, CD3 + ; B cells, CD20 + ; NK cells, CD56 + ; neutrophils, MRP14 + /CD66b + ; monocytes, MRP14 + /CCR2 + ; macrophages, MRP14 + /HLA-DR + ). (G) Relative proportion (of total CD45 + cells) of cell types in all 19 FOVs (left), and average cell numbers (summary, right).

Article Snippet: CD66b-PE , Miltenyi Biotec , Cat# 130-122-922, N/A.

Techniques: Computed Tomography, RNA Sequencing Assay, Immunohistochemistry, Immunofluorescence, Microscopy, Electron Microscopy, MANN-WHITNEY, Fluorescence, In Situ Hybridization

Journal: Cell

Article Title: SARS-CoV-2 infection triggers profibrotic macrophage responses and lung fibrosis

doi: 10.1016/j.cell.2021.11.033

Figure Lengend Snippet:

Article Snippet: CD66b-PE , Miltenyi Biotec , Cat# 130-122-922, N/A.

Techniques: Immunohistochemistry, Plasmid Preparation, Recombinant, Staining, Lysis, Protease Inhibitor, Mass Spectrometry, Sequencing, Modification, Bicinchoninic Acid Protein Assay, Enzyme-linked Immunosorbent Assay, Software

Journal:

Article Title: Involvement of Toll-Like Receptor 4 in Innate Immunity to Respiratory Syncytial Virus

doi: 10.1128/JVI.75.22.10730-10737.2001

Figure Lengend Snippet: Impaired NK cell activation in TLR4null mice after RSV infection. BAL samples from TLR4null and TLR4wt mice were harvested 7 days post-RSV or -influenza virus (FLU) infection. NK lytic activity against YAC-1 target cells was determined. Results are expressed as the mean of three independent experiments ± the standard error of the mean. Asterisks indicate a significant difference (P < 0.05) between TLR4null and TLR4wt mice. E:T Ratio, effector-to-target ratio.

Article Snippet: YAC-1 cells (ATCC TIB 160) were used as target cells.

Techniques: Activation Assay, Infection, Activity Assay

Journal:

Article Title: Involvement of Toll-Like Receptor 4 in Innate Immunity to Respiratory Syncytial Virus

doi: 10.1128/JVI.75.22.10730-10737.2001

Figure Lengend Snippet: RSV-induced NK cytotoxicity is perforin dependent. Seven days p.i., BAL samples from TLR4null (open bar) and TLR4wt (closed bar) mice were examined following RSV infection. Samples were purified for NK cells (DX5+) (80 to 90% enrichment) by positive selection using the MACS separation system. NK lytic activity against YAC-1 target cells was assessed in the presence or absence of EGTA-MgCl2+ at effector-to-target ratios of 40:1 (presented), 20:1, 10:1, and 5:1 (data not shown). The asterisk indicates a significant difference (P < 0.05) between TLR4null and TLR4wt mice. The results are representative of two independent experiments.

Article Snippet: YAC-1 cells (ATCC TIB 160) were used as target cells.

Techniques: Infection, Purification, Selection, Activity Assay

Journal:

Article Title: Involvement of Toll-Like Receptor 4 in Innate Immunity to Respiratory Syncytial Virus

doi: 10.1128/JVI.75.22.10730-10737.2001

Figure Lengend Snippet: Addition of IL-12 enhances NK-mediated cytotoxicity in RSV-infected TLR4null mice. BAL samples from TLR4null and TLR4wt mice were examined for cytotoxicity 7 days post-RSV infection. Effector BAL cells were cultured in the presence of 2 ng of IL-12/ml for 24 h. NK lytic activities against YAC-1 target cells at effector-to-target ratios of 40:1 (presented), 20:1, 10:1, and 5:1 (data not shown) were analyzed. A representative experiment is shown.

Article Snippet: YAC-1 cells (ATCC TIB 160) were used as target cells.

Techniques: Infection, Cell Culture