Journal: Journal of Veterinary Internal Medicine

Article Title: Matrix metalloproteinase‐2, ‐7, and ‐9 activities in dogs with idiopathic pulmonary fibrosis compared to healthy dogs and dogs with other respiratory diseases

doi: 10.1111/jvim.15970

Figure Lengend Snippet: A, A representative image of a caseinolytic zymogram of serum samples. White bands of a West Highland white terrier (WHWT) with canine idiopathic pulmonary fibrosis (CIPF) and a healthy WHWT indicate pro‐matrix metalloproteinase (MMP)‐7 activity. No activity is detected in a sample from a healthy dog of other breed. B, A representative image of a gelatinolytic zymogram of bronchoalveolar lavage fluid. Both pro‐MMP‐2 and pro‐MMP‐9 activity are detected in the samples of a dog with chronic bronchitis (CB) and a WHWT with CIPF. Active MMP‐9 activity is present only in the sample of the CIPF WHWT

Article Snippet: To identify the caseinolytic band as pro‐MMP‐7, a Western blot assay was performed using MMP‐7 polyclonal antibody (Bioss Antibodies, dilution 1:300 and 1:1000) and horseradish peroxidase‐conjugated goat anti‐rabbit immunoglobulin (Dako, dilution 1:2000) for 2 serum samples.

Techniques: Activity Assay

Journal: Journal of Veterinary Internal Medicine

Article Title: Matrix metalloproteinase‐2, ‐7, and ‐9 activities in dogs with idiopathic pulmonary fibrosis compared to healthy dogs and dogs with other respiratory diseases

doi: 10.1111/jvim.15970

Figure Lengend Snippet: Scatter plot (median and interquartile range) of serum pro‐matrix metalloproteinase‐7 activities in West Highland white terriers with canine idiopathic pulmonary fibrosis compared with other groups. BP, bacterial pneumonia; CB, chronic bronchitis; CIPF, canine idiopathic pulmonary fibrosis, EBP, eosinophilic bronchopneumopathy; WHWT, West Highland white terrier

Article Snippet: To identify the caseinolytic band as pro‐MMP‐7, a Western blot assay was performed using MMP‐7 polyclonal antibody (Bioss Antibodies, dilution 1:300 and 1:1000) and horseradish peroxidase‐conjugated goat anti‐rabbit immunoglobulin (Dako, dilution 1:2000) for 2 serum samples.

Techniques:

Journal: International Journal of Nanomedicine

Article Title: NanoVelcro-captured CTC number concomitant with enhanced serum levels of MMP7 and MMP9 enables accurate prediction of metastasis and poor prognosis in patients with lung adenocarcinoma

doi: 10.2147/IJN.S144033

Figure Lengend Snippet: Correlation between CTC counts and clinicopathologic characteristics of LADC

Article Snippet: Serum concentrations of MMP7 and MMP9 were quantified by ELISA using the MMP7, MMP9 ELISA kit (Boster, People’s Republic of China), respectively.

Techniques: Expressing

Journal: International Journal of Nanomedicine

Article Title: NanoVelcro-captured CTC number concomitant with enhanced serum levels of MMP7 and MMP9 enables accurate prediction of metastasis and poor prognosis in patients with lung adenocarcinoma

doi: 10.2147/IJN.S144033

Figure Lengend Snippet: MMP7 and MMP9 correlate with CTC count and poor clinical prognosis. Notes: ( A ) Serum levels of MMP7 and MMP9 were significantly increased in LADC patients with metastasis compared to those without metastasis (** p <0.01). ( B , C ) Remarkable associations were observed between pTNM stage and MMP7 and MMP9 expression levels (** p <0.01). ( D , E ) CTC counts were positively correlated with serum MMP7 ( r =0.892; p <0.01) and MMP9 ( r =0.891; p <0.01) expression levels. Abbreviations: CTC, circulating tumor cell; MMP7/9, matrix metalloproteinase 7/9; MOD, mean optical density.

Article Snippet: Serum concentrations of MMP7 and MMP9 were quantified by ELISA using the MMP7, MMP9 ELISA kit (Boster, People’s Republic of China), respectively.

Techniques: Expressing

Journal: International Journal of Nanomedicine

Article Title: NanoVelcro-captured CTC number concomitant with enhanced serum levels of MMP7 and MMP9 enables accurate prediction of metastasis and poor prognosis in patients with lung adenocarcinoma

doi: 10.2147/IJN.S144033

Figure Lengend Snippet: MMP7 and MMP9 inhibit adhesion and promote LADC cell migration and invasion in vitro. Notes: ( A ) Stable overexpression or knockdown of endogenous MMP7/MMP9 in LADC cell lines (NCI-H1975 and NCI-H2122) was confirmed by Western blotting. ( B ) MMP7 and MMP9 overexpression inhibited cell adhesion; MMP7 and MMP9 depletion increased cell adhesion. ( C , D ) Ectopic overexpression of MMP7 and MMP9 promoted cell invasion (** p <0.01); knockdown of endogenous MMP7 and MMP9 inhibited cell invasion (** p <0.01). * p <0.05. Abbreviations: LADC, lung adenocarcinoma; MMP7/9, matrix metalloproteinase 7/9.

Article Snippet: Serum concentrations of MMP7 and MMP9 were quantified by ELISA using the MMP7, MMP9 ELISA kit (Boster, People’s Republic of China), respectively.

Techniques: Migration, In Vitro, Over Expression, Knockdown, Western Blot

Journal: International Journal of Nanomedicine

Article Title: NanoVelcro-captured CTC number concomitant with enhanced serum levels of MMP7 and MMP9 enables accurate prediction of metastasis and poor prognosis in patients with lung adenocarcinoma

doi: 10.2147/IJN.S144033

Figure Lengend Snippet: MMP7 and MMP9 facilitate the liberation of LADC cells into the bloodstream in vivo. Notes: ( A ) Bioluminescence imaging of NOD/SCID mice 30 days after subcutaneous injection of different versions of NCI-H1975 cells and control cells. ( B ) MMP7 and MMP9 overexpression decreased CTC levels; MMP7 and MMP9 depletion increased CTC count. Abbreviations: CTC, circulating tumor cell; LADC, lung adenocarcinoma; MMP7/9, matrix metalloproteinase 7/9.

Article Snippet: Serum concentrations of MMP7 and MMP9 were quantified by ELISA using the MMP7, MMP9 ELISA kit (Boster, People’s Republic of China), respectively.

Techniques: In Vivo, Imaging, Injection, Control, Over Expression

Journal: International Journal of Nanomedicine

Article Title: NanoVelcro-captured CTC number concomitant with enhanced serum levels of MMP7 and MMP9 enables accurate prediction of metastasis and poor prognosis in patients with lung adenocarcinoma

doi: 10.2147/IJN.S144033

Figure Lengend Snippet: Metastatic significance of concomitant CTC count, serum MMP7 expression, and serum MMP9 expression. Notes: The cutoff values for CTC number, serum MMP7 level, and serum MMP9 level had great sensitivity and specificity for metastasis, with an area under the curve (AUC) of ( A ) 0.945 (95% CI: 0.900–0.990; p <0.01), ( B ) 0.949 (95% CI: 0.913–0.986; p <0.01), and ( C ) 0.930 (95% CI: 0.881–0.979; p <0.01), respectively. ( D ) The predicted cutoff value of the combination of CTC count, serum MMP7 level, and serum MMP9 level had an optimal AUC of 0.950 (95% CI: 0.909–0.992; p <0.01). Abbreviations: CTC, circulating tumor cell; MMP7/9, matrix metalloproteinase 7/9.

Article Snippet: Serum concentrations of MMP7 and MMP9 were quantified by ELISA using the MMP7, MMP9 ELISA kit (Boster, People’s Republic of China), respectively.

Techniques: Expressing

Journal: International Journal of Nanomedicine

Article Title: NanoVelcro-captured CTC number concomitant with enhanced serum levels of MMP7 and MMP9 enables accurate prediction of metastasis and poor prognosis in patients with lung adenocarcinoma

doi: 10.2147/IJN.S144033

Figure Lengend Snippet: Increased CTC count, serum MMP7 level, and serum MMP9 level determine the worst prognosis. Notes: ( A ) Kaplan–Meier curves revealed the shortest overall survival (OS). ( B ) The survival analysis revealed the shortest median survival and a lower 5-year survival rate. Abbreviations: CTC, circulating tumor cell; MMP7/9, matrix metalloproteinase 7/9.

Article Snippet: Serum concentrations of MMP7 and MMP9 were quantified by ELISA using the MMP7, MMP9 ELISA kit (Boster, People’s Republic of China), respectively.

Techniques:

Journal: International Journal of Nanomedicine

Article Title: NanoVelcro-captured CTC number concomitant with enhanced serum levels of MMP7 and MMP9 enables accurate prediction of metastasis and poor prognosis in patients with lung adenocarcinoma

doi: 10.2147/IJN.S144033

Figure Lengend Snippet: Univariate and multivariate analyses of overall survival in LADC patients

Article Snippet: Serum concentrations of MMP7 and MMP9 were quantified by ELISA using the MMP7, MMP9 ELISA kit (Boster, People’s Republic of China), respectively.

Techniques: Expressing

Journal: International Journal of Nanomedicine

Article Title: NanoVelcro-captured CTC number concomitant with enhanced serum levels of MMP7 and MMP9 enables accurate prediction of metastasis and poor prognosis in patients with lung adenocarcinoma

doi: 10.2147/IJN.S144033

Figure Lengend Snippet: siRNA sequences used in this study

Article Snippet: Serum concentrations of MMP7 and MMP9 were quantified by ELISA using the MMP7, MMP9 ELISA kit (Boster, People’s Republic of China), respectively.

Techniques:

Journal: International Journal of Nanomedicine

Article Title: NanoVelcro-captured CTC number concomitant with enhanced serum levels of MMP7 and MMP9 enables accurate prediction of metastasis and poor prognosis in patients with lung adenocarcinoma

doi: 10.2147/IJN.S144033

Figure Lengend Snippet: PCR primers used in this study

Article Snippet: Serum concentrations of MMP7 and MMP9 were quantified by ELISA using the MMP7, MMP9 ELISA kit (Boster, People’s Republic of China), respectively.

Techniques:

Journal: Advanced functional materials

Article Title: Matrix Metalloproteinase Responsive, Proximity-activated Polymeric Nanoparticles for siRNA Delivery

doi: 10.1002/adfm.201202215

Figure Lengend Snippet: Synthetic scheme of MMP-7-responsive, PEG-pep-pD-pDPB (forming PAT-SPN) polymer construct.

Article Snippet: Synthesis of PEG-peptide RAFT macro chain transfer agent The MMP-7-cleavable peptide (H- VPLS↓LYSG CG-OH; arrow indicates expected MMP-7 cleavage site) was synthesized through standard Fmoc-based solid phase techniques on a PS3 synthesizer (Protein Technologies, Tucson, AZ).

Techniques: Construct

Journal: Advanced functional materials

Article Title: Matrix Metalloproteinase Responsive, Proximity-activated Polymeric Nanoparticles for siRNA Delivery

doi: 10.1002/adfm.201202215

Figure Lengend Snippet: PEG-pep-pD-pDPB generated from PEG-peptide macro-CTA (dash-dot) after two successive RAFT polymerizations was confirmed by the shift in GPC elution time (dash-dot versus solid line traces). Elugrams of PEG-pep-pD-pDPB treated with MMP-7 (dash) was consistent with cleavage of the distal PEG segment as indicated by the shift of the primary elution peak (solid versus dashed traces at ∼23 min) and the appearance of a shoulder peak at ∼25 min that corresponds to the PEG-peptide macro-CTA (dash-dot).

Article Snippet: Synthesis of PEG-peptide RAFT macro chain transfer agent The MMP-7-cleavable peptide (H- VPLS↓LYSG CG-OH; arrow indicates expected MMP-7 cleavage site) was synthesized through standard Fmoc-based solid phase techniques on a PS3 synthesizer (Protein Technologies, Tucson, AZ).

Techniques: Generated

Journal: Advanced functional materials

Article Title: Matrix Metalloproteinase Responsive, Proximity-activated Polymeric Nanoparticles for siRNA Delivery

doi: 10.1002/adfm.201202215

Figure Lengend Snippet: Nanocarrier zeta potential increased from +5.8 to +14.4 mV over 6.5 h of treatment with a physiologically-relevant MMP-7 concentration (50 nM).

Article Snippet: Synthesis of PEG-peptide RAFT macro chain transfer agent The MMP-7-cleavable peptide (H- VPLS↓LYSG CG-OH; arrow indicates expected MMP-7 cleavage site) was synthesized through standard Fmoc-based solid phase techniques on a PS3 synthesizer (Protein Technologies, Tucson, AZ).

Techniques: Concentration Assay

Journal: Advanced functional materials

Article Title: Matrix Metalloproteinase Responsive, Proximity-activated Polymeric Nanoparticles for siRNA Delivery

doi: 10.1002/adfm.201202215

Figure Lengend Snippet: (a-b) Flow cytometry analysis of DNAFAM delivered to MDA-MB-231 breast cancer cells using intact PAT-SPNs (PAT-SPN/DNAFAM, blue), MMP-7-pretreated PAT-SPNs (MMP7 pretreated PAT-SPN/DNAFAM, yellow), or no treatment (NT) (red). Mean FAM fluorescence was significantly higher in cells treated with PAT-SPN/DNAFAM proximity activated by MMP-7 compared to all other treatment groups. Data presented as mean +/- standard error with n=3. *p < 0.05 compared with NT and #p < 0.05 compared with PAT-SPN/FAM-DNA. (c) Confocal microscopy images of MDA-MB-231 breast cancer cells treated with PAT-SPN/DNACy5 nanocarriers (at 50 and 100 nM DNA concentration) with or without MMP-7 treatment correlated with quantitative flow cytometry data. PAT-SPN/DNA complexes of N(+)/P(-) = 6 were applied.

Article Snippet: Synthesis of PEG-peptide RAFT macro chain transfer agent The MMP-7-cleavable peptide (H- VPLS↓LYSG CG-OH; arrow indicates expected MMP-7 cleavage site) was synthesized through standard Fmoc-based solid phase techniques on a PS3 synthesizer (Protein Technologies, Tucson, AZ).

Techniques: Flow Cytometry, Fluorescence, Confocal Microscopy, Concentration Assay

Journal: Advanced functional materials

Article Title: Matrix Metalloproteinase Responsive, Proximity-activated Polymeric Nanoparticles for siRNA Delivery

doi: 10.1002/adfm.201202215

Figure Lengend Snippet: (a) pH-dependent hemolysis of erythrocytes was determined for intact PAT-SPN and MMP-7 pretreated PAT-SPN at 1, 5, 15, and 30 μg/mL relative to 1% Triton X-100 as a positive control (100% lysis). Hemolysis was pH-dependent, with approximately 0% at extracellular pH 7.4 to more than 90% at pH 5.6. Hemolysis mediated by PAT-SPN is essentialy unchanged following MMP-7 proteolytic activation. Error bars indicate standard deviation of n = 4. (b) Confocal imaging of MDA-MB-231 cells treated with MMP-7 pretreated PAT-SPN/DNACy5 and stained with LysoTracker. Control cells only stained by LysoTracker are shown as a reference (upper panel). Intracellular DNACy5 delivered by the MMP-7 pretreated PAT-SPNs is predominantly distributed outside the lysosomes (red) with minimal colocalization with the LysoTracker signal (yellow).

Article Snippet: Synthesis of PEG-peptide RAFT macro chain transfer agent The MMP-7-cleavable peptide (H- VPLS↓LYSG CG-OH; arrow indicates expected MMP-7 cleavage site) was synthesized through standard Fmoc-based solid phase techniques on a PS3 synthesizer (Protein Technologies, Tucson, AZ).

Techniques: Positive Control, Lysis, Activation Assay, Standard Deviation, Imaging, Staining

Journal: Advanced functional materials

Article Title: Matrix Metalloproteinase Responsive, Proximity-activated Polymeric Nanoparticles for siRNA Delivery

doi: 10.1002/adfm.201202215

Figure Lengend Snippet: (a) Protein level gene silencing readouts were acquired in R221A-Luc mammary tumor cells that constitutively express firefly luciferase, following treatment with luciferase-targeted siRNA complexed into PAT-SPNs at N(+)/P(-) charge ratios of 2, 4, and 6. Lipofectamine (Lipofec) was used as a positive control, and luciferase activity was reported relative to untreated cells (NT). (Data presented as mean +/- standard error with n=3). *p < 0.05 compared with NT, #p < 0.05 compared with Lipofec, and $p < 0.001 between the intact and the MMP-7 pretreated PAT-SPN groups. (b) Variance analysis by ANOVA(one way Tukey's range test) on knockdown results of PAT-SPN/siRNA versus no treatment (NT) (left) or versus Lipofectamine™/siRNA (Lipofec) (right). Formulations with variance > 0 are significantly different to NT (left) and to Lipofec (right)

Article Snippet: Synthesis of PEG-peptide RAFT macro chain transfer agent The MMP-7-cleavable peptide (H- VPLS↓LYSG CG-OH; arrow indicates expected MMP-7 cleavage site) was synthesized through standard Fmoc-based solid phase techniques on a PS3 synthesizer (Protein Technologies, Tucson, AZ).

Techniques: Luciferase, Positive Control, Activity Assay