Journal: Emerging Infectious Diseases

Article Title: Livestock Susceptibility to Infection with Middle East Respiratory Syndrome Coronavirus

doi: 10.3201/eid2302.161239

Figure Lengend Snippet: Presence of MERS-CoV receptor DPP4 (IHC) and of mucosubstances (PAS) in upper and lower respiratory tract tissues from sheep, pigs, llamas, and horses. A) In the nose, DPP4 (red cytoplasmic or membrane staining) was present on the lining epithelium of pigs, llamas, and horses but not sheep. PAS staining (magenta) demonstrated more mucous cells in the lining epithelium of sheep and horses and a layer of mucus on the lining epithelium of the horses. B) DPP4 (red cytoplasmic or membrane staining) was present on the lining epithelium of the trachea, bronchus/bronchioles, and alveoli in the pigs, llamas and horses but not in the sheep. Original magnification ×400 for all images. DPP4, dipeptidyl peptidase-4; IHC, immunohistochemistry; MERS-CoV, Middle East respiratory syndrome coronavirus; PAS, periodic acid–Schiff; term., terminal.

Article Snippet: In brief, we performed DPP4 IHC staining by using 5 μg/mL of polyclonal goat IgG anti-human DPP4 antibody (R&D Systems, Abingdon, UK) and peroxidase-labeled rabbit anti-goat IgG (1:200; DAKO; Agilent Technologies Company, Santa Clara, CA, USA) as a secondary antibody.

Techniques: Membrane, Staining, Immunohistochemistry

Journal: Journal of Pediatric Hematology/Oncology

Article Title: Chemotherapy-induced Alteration of SDF-1/CXCR4 Expression in Bone Marrow–derived Mesenchymal Stem Cells From Adolescents and Young Adults With Acute Lymphoblastic Leukemia

doi: 10.1097/mph.0000000000000220

Figure Lengend Snippet: FIGURE 3. Secretion and expression of SDF-1 in BM-MSCs. A, SDF-1 level was measured by enzyme-linked immunosorbent assay. (a) The bone marrow plasma level of SDF-1 in patients with ALL was significantly increased compared with that in healthy donors and ALL-c (*P < 0.05; n = 10). (b), The same difference was found when SDF-1 levels were measured in the supernatants of MSC cultures after incubation for 72 hours. B, Western blotting was performed to quantify total SDF-1 in the lysates of MSCs. SDF-1 expression in the MSCs of ALL was significantly higher than that in the MSCs of healthy donors and ALL-c. C, The difference was also found at the transcriptional level by real-time PCR (*P < 0.05, n = 10). D, Surface-CD26 expression on the MSCs from ALL patients measured by flow cytometry was significantly lower than that in healthy donors and ALL-c (*P < 0.05; n = 10). E, Total CD26 was measured by Western blot in the MSCs. The fold difference was shown among the 3 types of MSCs. F, The same difference was also found at the transcriptional level (*P < 0.05; n = 10). ALL indicates acute lymphoblastic leukemia; BM-MSC, bone marrow–derived mesenchymal stem cell; GADPH, glyceraldehyde- 3-phosphate dehydrogenase; PCR, polymerase chain reaction.

Article Snippet: Membranes were hybridized with the following primary antibodies: rabbit anti-human CXCR4 (1:1000; Abcam), goat antihuman SDF-1(1:500; R&D Systems), goat anti-human CD26 (1:1000; R&D Systems), and mouse antibody to glyceraldehyde-3-phosphate dehydrogenase (1:4000; Santa Cruz).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Incubation, Western Blot, Real-time Polymerase Chain Reaction, Flow Cytometry, Derivative Assay, Polymerase Chain Reaction

Journal: Hypertension

Article Title: Dipeptidyl Peptidase IV Regulates Proliferation of Preglomerular Vascular Smooth Muscle and Mesangial Cells

doi: 10.1161/hypertensionaha.112.196501

Figure Lengend Snippet: Figure 1. A, Dipeptidyl peptidase IV (DPPIV) mRNA levels in spontaneously hypertensive rat (SHR) and Wistar-Kyoto rat (WKY) preglomerular microvascular smooth muscle cells (PGVSMCs) and glomerular mesangial cells (GMCs). DPPIV mRNA expression was determined by quantitative real-time PCR and normalized to -actin mRNA. B, Western blotting for DPPIV protein using 2 different antibodies (Ab 1 and 2). A dis- tinct and single band was observed at 55 kDa for both WKY and SHR PGVSMCs and GMCs. C, DPPIV protein expression in SHR and WKY PGVSMCs and GMCs. DPPIV protein expression was quantified by densitometry analysis of Western blots and normalized to -actin protein. Letter above bars (a) indicates significantly different (Fisher least significant difference [LSD] test) from PGVSMCs.

Article Snippet: To ensure that the signal detected by Western blotting was indeed DPPIV, we performed Western blotting for DPPIV protein as described in Jackson et al.3 using two different primary antibodies: 1) rabbit polyclonal anti-DPPIV antibody (Proteintech Group, Inc.; Chicago, IL; catalog No. 10940-1-AP; 1:600); and 2) rabbit polyclonal anti-DPPIV antibody (Thermo Scientific, Rockford, IL; catalogue number PA1-8455; 1:2500).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: International Journal of Cardiology

Article Title: Cardiac inflammation and microvascular procoagulant changes are decreased in second wave compared to first wave deceased COVID-19 patients

doi: 10.1016/j.ijcard.2021.11.079

Figure Lengend Snippet: Comparison of coagulation regulating factors and CML in the cardiac microvasculature between first and second wave COVID-19 and control patients. The number of blood vessels positive for the procoagulant factors Tissue Factor (A; TF), Factor VII (B; FVII), Factor XII (C; FXII) and anticoagulant dipeptidyl peptidase 4 (D; DPP4) are shown per cm 2 of left ventricular heart tissue in control patients (Con; n = 18) and first wave (Wave 1; n = 15) and second wave (Wave 2; n = 10) COVID-19 patients. (E): An immunohistochemical example of the presence N(ε)-Carboxymethyllysine (CML) in the endothelium of intramyocardial blood vessels of a COVID-19 patient (arrow) and the immunohistochemical (IH) score for CML per cm 2 in control patients (Con) and first- and second wave COVID-19 patients. (F): The number of blood vessels with weak, moderate and strong CML staining (staining intensities 1, 2, 3 respectively) in control (Con) and first- and second wave COVID-19 patients. Each point in the graphs represents the value of one individual patient, the bars represent mean ± SD. The bars represent mean ± SD. For comparisons Kruskal-Wallis tests with Dunn's multiple comparison tests were used. *p < 0.05, **p < 0.01, *** p < 0.001, **** p < 0.0001 (exact p-values are given in the text).

Article Snippet: Primary antibodies were added for 1 h at room temperature (RT): mouse-anti-human CD45 (1:100, Dako Santa Clara, USA; M0701), rabbit-anti-human CD68 (1:400, Dako; M0814), rabbit-anti-human CD3 (1:100, Dako; A0452), rabbit-anti-human C3d (1:1000, Dako; A0063), mouse-anti-human CD31 (1:50, Dako; M0823), mouse-anti-human TF (1:250, Biorbyt Cambridge, UK; ORB100189), mouse-anti-human FVII (1:100, Sanquin Research, Amsterdam, The Netherlands), mouse-anti-human FXII (1:25, Sanquin), mouse-anti-human CML (1:500 [ ]) or mouse-anti-human CD26 (1:100, Bio-Rad, Lunteren, The Netherlands, MCA1317T).

Techniques: Comparison, Coagulation, Control, Immunohistochemical staining, Staining