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1.
Figure 7

Figure 7. Ad5-TTP reduces osteoclastogenesis in experimental bone loss model. From: Targeting mRNA Stability Arrests Inflammatory Bone Loss.

Tartate-resistant acid phosphatase–positive (TRAP+) cells were enumerated from similar sagitally sectioned specimens and graphically presented as a scatter plot analysis. Horizontal bar indicates mean TRAP+ cell counts (n = 8; ***P < 0.001). Ad, adenovirus; GFP, green fluorescent protein; TTP, tristetraprolin.

Chetan S Patil, et al. Mol Ther. ;16(10):1657-1664.
2.
Figure 6

Figure 6. Ad5-TTP reduces inflammatory infiltrate induced by Aggregatibacter actinomycetemcomitans lipopolysaccharide (LPS) in the area adjacent to bone loss. From: Targeting mRNA Stability Arrests Inflammatory Bone Loss.

Histological appearance of representative phosphate-buffered saline (PBS)-injected control (a), PBS + Ad5-GFP (b), PBS + Ad5-TTP (c), LPS (d), LPS + Ad5-GFP (e), LPS + Ad5-TTP (f) at low magnification (bars indicate 1,000 µm). Inserts of boxed areas between M1 and M2 at higher magnification (bars = 40 µm). Ad, adenovirus; GFP, green fluorescent protein; TTP, tristetraprolin.

Chetan S Patil, et al. Mol Ther. ;16(10):1657-1664.
3.
Figure 4

Figure 4. Ad5-TTP is expressed in periodontal tissues throughout the experimental protocol. From: Targeting mRNA Stability Arrests Inflammatory Bone Loss.

Immunohistochemical analysis of tristetraprolin (TTP) expression in adenovirus-transduced periodontal tissues from (a) Ad5-GFP and (d) Ad5-TTP at low magnification. (b and e) Upper boxed region from a and d, respectively, at higher magnification showing Ad5-GFP-injected area between maxillary molars areas. Larger arrows indicate TTP expression in epithelial tissue and shorter arrows indicate connective tissue staining. (c and f) Lower boxed region from a and c, respectively, at higher magnification showing adjacent nontransduced periodontal tissues. Bars indicate scale. Ad, adenovirus; GFP, green fluorescent protein.

Chetan S Patil, et al. Mol Ther. ;16(10):1657-1664.
4.
Figure 5

Figure 5. Ad5-TTP protects bone in experimental periodontal disease model. From: Targeting mRNA Stability Arrests Inflammatory Bone Loss.

(a) Representative microcomputed tomography images of rat maxillae from indicated treatment groups. (b) Linear measurement from cementoenamel junction (CEJ) to alveolar bone crest (ABC) (n = 8/group; *P < 0.05, **P < 0.01). (c) Volumetric analysis of bone loss levels (n = 8/group; *P < 0.05, **P < 0.01). Ad, adenovirus; GFP, green fluorescent protein; PBS, phosphate-buffered saline; TTP, tristetraprolin.

Chetan S Patil, et al. Mol Ther. ;16(10):1657-1664.
5.
Figure 1

Figure 1. Ad5-TTP potently decreases secretion of inflammatory mediators in macrophages. From: Targeting mRNA Stability Arrests Inflammatory Bone Loss.

RAW264.7 macrophages were transduced with Ad5-GFP or Ad5-TTP and stimulated with Aggregatibacter actinomycetemcomitans lipopolysaccharide (LPS) (1 µg/ml). Cell culture supernatants were obtained 24 hours after stimulation and subjected to enzyme-linked immunosorbent assay. Ad5-TTP reduced LPS-induced (a) tumor necrosis factor-α (TNF-α) (***P < 0.001), (b) interleukin-6 (IL-6) (***P < 0.001), and (c) prostaglandin (PG)E2 (***P < 0.001; n = 3 for each). Ad, adenovirus; GFP, green fluorescent protein; TTP, tristetraprolin.

Chetan S Patil, et al. Mol Ther. ;16(10):1657-1664.
6.
Figure 3

Figure 3. Ad5-luciferase is locally expressed in rat maxillae for up to 21 days. From: Targeting mRNA Stability Arrests Inflammatory Bone Loss.

Rats were injected in the left maxillae with Ad5-luciferase reporter gene. Rats were the given luciferin substrate on days 1, 3, 7, 14, and 21 and in vivo bioluminescence was measured. (a) Representative images of rat maxillae at day 7 at indicated multiplicities of infection (MOIs) with Ad5-luciferase. (b) Graphical presentation of luciferase activity over 21 days. (n = 3/MOI). ROI, region of interest.

Chetan S Patil, et al. Mol Ther. ;16(10):1657-1664.
7.
Figure 2

Figure 2. Ad5-TTP specifically inhibits adenylate-uridylate-rich element (ARE)-containing transcripts. From: Targeting mRNA Stability Arrests Inflammatory Bone Loss.

(a) RAW264.7 macrophages were transduced with Ad5-GFP or Ad5-TTP and stimulated with Aggregatibacter actinomycetemcomitans lipopolysaccharide (LPS) (1 µg/ml). RNA was isolated 18 hours after stimulation and subjected to real-time reverse transcription-PCR measurement of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and cyclooxygenase-2 (COX-2) normalized to glyceraldehyde 3-phosphate dehydrogenase mRNA (n = 3 each). (b) HeLa cells were “adenofected” with luciferase reporter constructs and Ad5-TTP [75 multiplicity of infection (MOI)], Ad5-GFP (75 MOI), or no adenovirus (Ad). Luciferase reporters contained 3′-untranslated region (3′-UTR) of TNF-α, IL-6, and COX-2, or non-ARE control pGL3-control. Luciferase activity was measured and normalized to total protein (n = 3; *P < 0.05, **P < 0.01, ***P < 0.001). TTP, tristetraprolin.

Chetan S Patil, et al. Mol Ther. ;16(10):1657-1664.

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