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  • 93
    Durect Corporation osmotic minipumps
    T cell survival is mediated by DC-secreted factors and IL-6. (A) Expression of GM-CSFR-α, GM-CSFR-β (AIC2B), and IL-2Rβ was determined by quantitative RT-PCR using cDNA of cells indicated. Naive CD62L + CD4 + T cells (lane 1) and splenic CD11c + DC (lane 5) were purified by flow cytometry (purity > 98%). Th1 (lane 2), Th2 (lane 3), and Th17 (lane 4) polarized CD4 + T cells. (B and C) DC of GM-CSF −/− or GM-CSF +/+ mice were cultured with purified CD4 + T cells from DO11.10/GM-CSF −/− or DO11.10/GM-CSF +/+ mice, respectively, in the presence of titrating amounts of OVA 323-339. (B) IL-6 levels in the culture supernatant at day 3 were determined by ELISA. (C) Proliferation measured by 3 H-Thymidine incorporation in the absence and in the presence of 20 ng/ml rIL-6. (D) Splenocytes of DO11.10/GM-CSF +/+ or GM-CSF −/− mice were cultured with 1 μM OVA 323-339 in the absence and presence of 20ng/ml rIL-6 for 3 d before staining of cells with KJ1-26 + mAb, Annexin V (AV), and PI and analysis by flow cytometry. Dot plot gated on KJ1-26 + cells shows early apoptotic cells (PI − AV + ), late apoptotic (PI + AV + ), and dead cells (PI + AV − ). (E) GM-CSF–deficient cultures were supplemented with rIL-1, rIL-2, or rIL-6. Proliferation was measured by 3 H-Thymidine incorporation after 3 d of culture. Proliferation index was calculated as described in Materials and methods. (F) CD4 + T cells purified from DO11.10/GM-CSF +/+ or DO11.10/GM-CSF −/− mice were injected i.v. into GM-CSF +/+ or GM-CSF −/− mice, respectively. 2 d later, groups of KO and WT mice were implanted with osmotic <t>minipumps</t> containing hIL-6 or were sham operated. Subsequently, mice were immunized with 200 μg OVA 323-339 peptide emulsified in CFA. After 7 d, draining LN and spleen cells were analyzed by flow cytometry. Symbols indicate total number of splenic KJ1-26 + cells of individual mice. Horizontal lines indicate averages of groups. Shown is one representative experiment of three performed. *,
    Osmotic Minipumps, supplied by Durect Corporation, used in various techniques. Bioz Stars score: 93/100, based on 1737 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ALZA osmotic minipumps
    Recovery of weight bearing after NgR1 treatment of chronically spinal cord contused rats A , Spontaneous improvement of open field locomotion (BBB score) as a function of time after thoracic spinal cord contusion injury for all rats prior to randomization to IgG or NgR1 treatment. Mean ± sem, n = 64. B , Schematic of experiment. Two weeks after the i.c.v. cannula implantation (12 weeks post-contusion injury), rats were assigned to one of two treatment groups. The PBS <t>minipumps</t> were replaced with new osmotic minipumps filled with 2.25 mg AA-NgR(310)ecto-Fc (0.29 mg/kg/day) or 2.25 mg rat IgG in 2 ml PBS. The duration of treatment was 12 weeks. A new osmotic mimipump filled with same amount of AA-NgR(310)ecto-Fc or rat IgG replaced each depleted pump every 4 weeks. C , Examples of a control rat without weight support at the end of the treatment period and two of the seven AA-NgR(310)ecto-Fc treated rats that regained weight support. D , The increase in the percentage of rats showing body weight support with at least one hindlimb as a function of time during therapy is reported. P =0.022 by repeated measures ANOVA for the effect of NgR1 versus IgG treatment, and *, P
    Osmotic Minipumps, supplied by ALZA, used in various techniques. Bioz Stars score: 92/100, based on 81 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Durect Corporation 1004 osmotic minipumps
    Recovery of weight bearing after NgR1 treatment of chronically spinal cord contused rats A , Spontaneous improvement of open field locomotion (BBB score) as a function of time after thoracic spinal cord contusion injury for all rats prior to randomization to IgG or NgR1 treatment. Mean ± sem, n = 64. B , Schematic of experiment. Two weeks after the i.c.v. cannula implantation (12 weeks post-contusion injury), rats were assigned to one of two treatment groups. The PBS <t>minipumps</t> were replaced with new osmotic minipumps filled with 2.25 mg AA-NgR(310)ecto-Fc (0.29 mg/kg/day) or 2.25 mg rat IgG in 2 ml PBS. The duration of treatment was 12 weeks. A new osmotic mimipump filled with same amount of AA-NgR(310)ecto-Fc or rat IgG replaced each depleted pump every 4 weeks. C , Examples of a control rat without weight support at the end of the treatment period and two of the seven AA-NgR(310)ecto-Fc treated rats that regained weight support. D , The increase in the percentage of rats showing body weight support with at least one hindlimb as a function of time during therapy is reported. P =0.022 by repeated measures ANOVA for the effect of NgR1 versus IgG treatment, and *, P
    1004 Osmotic Minipumps, supplied by Durect Corporation, used in various techniques. Bioz Stars score: 90/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Durect Corporation model 2004 osmotic minipumps
    Recovery of weight bearing after NgR1 treatment of chronically spinal cord contused rats A , Spontaneous improvement of open field locomotion (BBB score) as a function of time after thoracic spinal cord contusion injury for all rats prior to randomization to IgG or NgR1 treatment. Mean ± sem, n = 64. B , Schematic of experiment. Two weeks after the i.c.v. cannula implantation (12 weeks post-contusion injury), rats were assigned to one of two treatment groups. The PBS <t>minipumps</t> were replaced with new osmotic minipumps filled with 2.25 mg AA-NgR(310)ecto-Fc (0.29 mg/kg/day) or 2.25 mg rat IgG in 2 ml PBS. The duration of treatment was 12 weeks. A new osmotic mimipump filled with same amount of AA-NgR(310)ecto-Fc or rat IgG replaced each depleted pump every 4 weeks. C , Examples of a control rat without weight support at the end of the treatment period and two of the seven AA-NgR(310)ecto-Fc treated rats that regained weight support. D , The increase in the percentage of rats showing body weight support with at least one hindlimb as a function of time during therapy is reported. P =0.022 by repeated measures ANOVA for the effect of NgR1 versus IgG treatment, and *, P
    Model 2004 Osmotic Minipumps, supplied by Durect Corporation, used in various techniques. Bioz Stars score: 88/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Charles River Laboratories alzet osmotic minipumps
    Uncl-Sema3E inhibits angiogenesis and tumour growth in the RIP-Tag2 model <t>Alzet</t> <t>minipumps</t> were implanted in 12-week-old RT2 mice to release locally in the pancreas Uncl-Sema3E ( n = 14) or mock saline ( n = 12) over a period of 2 weeks; other mice were treated orally with sunitinib ( n = 8) for the same period (see Materials and Methods Section for details); two independent experiments were performed, yielding consistent results. Average global tumour burden per mice in different experimental conditions. Vessel area in treated tumours was estimated based on CD31 staining ( n = 6, per each experimental group). Vessel coverage with mural cells was assessed by double staining for NG2 and CD31 ( n = 6). Hypoxic areas in islet tumours were revealed by staining with pimonidazol (PIMO, see Materials and Methods Section; n = 6); the same sections were costained with the endothelial cell marker Endoglin-CD105, and with DAPI. Primary RT2 tumour invasiveness was assessed by morphometric analysis of H E stained sections, as previously reported (Pàez-Ribes et al, 2009 ). Representative images of the different groups are shown on the right. Uncl-Sema3E treated tumours often displayed a non-invasive phenotype: i.e. the tumour was ‘encapsulated’ and well separated from the surrounding exocrine tissue (delimited by a dotted line). In contrast, in sunitinib-treated mice the tumour invasive front was often extensively intercalated with the surrounding normal exocrine tissue. The graph on the left indicates the average fraction of encapsulated (IT), microinvasive (IC1) and highly invasive (IC2) carcinomas per mice in each experimental condition ( n = 8). We assessed the statistical significance of the different relative representation of each tumour histotype in drug-treated versus mock control mice; * p = 0.003; ** p = 0.007; § p = 6.1E−05; §§ p = 1.7E−06. Scale bars, throughout the entire figure: 100 µm (20 µm in magnified insets in panel C ).
    Alzet Osmotic Minipumps, supplied by Charles River Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    ALZA 2001 osmotic minipumps
    Uncl-Sema3E inhibits angiogenesis and tumour growth in the RIP-Tag2 model <t>Alzet</t> <t>minipumps</t> were implanted in 12-week-old RT2 mice to release locally in the pancreas Uncl-Sema3E ( n = 14) or mock saline ( n = 12) over a period of 2 weeks; other mice were treated orally with sunitinib ( n = 8) for the same period (see Materials and Methods Section for details); two independent experiments were performed, yielding consistent results. Average global tumour burden per mice in different experimental conditions. Vessel area in treated tumours was estimated based on CD31 staining ( n = 6, per each experimental group). Vessel coverage with mural cells was assessed by double staining for NG2 and CD31 ( n = 6). Hypoxic areas in islet tumours were revealed by staining with pimonidazol (PIMO, see Materials and Methods Section; n = 6); the same sections were costained with the endothelial cell marker Endoglin-CD105, and with DAPI. Primary RT2 tumour invasiveness was assessed by morphometric analysis of H E stained sections, as previously reported (Pàez-Ribes et al, 2009 ). Representative images of the different groups are shown on the right. Uncl-Sema3E treated tumours often displayed a non-invasive phenotype: i.e. the tumour was ‘encapsulated’ and well separated from the surrounding exocrine tissue (delimited by a dotted line). In contrast, in sunitinib-treated mice the tumour invasive front was often extensively intercalated with the surrounding normal exocrine tissue. The graph on the left indicates the average fraction of encapsulated (IT), microinvasive (IC1) and highly invasive (IC2) carcinomas per mice in each experimental condition ( n = 8). We assessed the statistical significance of the different relative representation of each tumour histotype in drug-treated versus mock control mice; * p = 0.003; ** p = 0.007; § p = 6.1E−05; §§ p = 1.7E−06. Scale bars, throughout the entire figure: 100 µm (20 µm in magnified insets in panel C ).
    2001 Osmotic Minipumps, supplied by ALZA, used in various techniques. Bioz Stars score: 88/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Durect Corporation 2001 osmotic minipumps
    Uncl-Sema3E inhibits angiogenesis and tumour growth in the RIP-Tag2 model <t>Alzet</t> <t>minipumps</t> were implanted in 12-week-old RT2 mice to release locally in the pancreas Uncl-Sema3E ( n = 14) or mock saline ( n = 12) over a period of 2 weeks; other mice were treated orally with sunitinib ( n = 8) for the same period (see Materials and Methods Section for details); two independent experiments were performed, yielding consistent results. Average global tumour burden per mice in different experimental conditions. Vessel area in treated tumours was estimated based on CD31 staining ( n = 6, per each experimental group). Vessel coverage with mural cells was assessed by double staining for NG2 and CD31 ( n = 6). Hypoxic areas in islet tumours were revealed by staining with pimonidazol (PIMO, see Materials and Methods Section; n = 6); the same sections were costained with the endothelial cell marker Endoglin-CD105, and with DAPI. Primary RT2 tumour invasiveness was assessed by morphometric analysis of H E stained sections, as previously reported (Pàez-Ribes et al, 2009 ). Representative images of the different groups are shown on the right. Uncl-Sema3E treated tumours often displayed a non-invasive phenotype: i.e. the tumour was ‘encapsulated’ and well separated from the surrounding exocrine tissue (delimited by a dotted line). In contrast, in sunitinib-treated mice the tumour invasive front was often extensively intercalated with the surrounding normal exocrine tissue. The graph on the left indicates the average fraction of encapsulated (IT), microinvasive (IC1) and highly invasive (IC2) carcinomas per mice in each experimental condition ( n = 8). We assessed the statistical significance of the different relative representation of each tumour histotype in drug-treated versus mock control mice; * p = 0.003; ** p = 0.007; § p = 6.1E−05; §§ p = 1.7E−06. Scale bars, throughout the entire figure: 100 µm (20 µm in magnified insets in panel C ).
    2001 Osmotic Minipumps, supplied by Durect Corporation, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Durect Corporation 2004 osmotic minipumps
    Uncl-Sema3E inhibits angiogenesis and tumour growth in the RIP-Tag2 model <t>Alzet</t> <t>minipumps</t> were implanted in 12-week-old RT2 mice to release locally in the pancreas Uncl-Sema3E ( n = 14) or mock saline ( n = 12) over a period of 2 weeks; other mice were treated orally with sunitinib ( n = 8) for the same period (see Materials and Methods Section for details); two independent experiments were performed, yielding consistent results. Average global tumour burden per mice in different experimental conditions. Vessel area in treated tumours was estimated based on CD31 staining ( n = 6, per each experimental group). Vessel coverage with mural cells was assessed by double staining for NG2 and CD31 ( n = 6). Hypoxic areas in islet tumours were revealed by staining with pimonidazol (PIMO, see Materials and Methods Section; n = 6); the same sections were costained with the endothelial cell marker Endoglin-CD105, and with DAPI. Primary RT2 tumour invasiveness was assessed by morphometric analysis of H E stained sections, as previously reported (Pàez-Ribes et al, 2009 ). Representative images of the different groups are shown on the right. Uncl-Sema3E treated tumours often displayed a non-invasive phenotype: i.e. the tumour was ‘encapsulated’ and well separated from the surrounding exocrine tissue (delimited by a dotted line). In contrast, in sunitinib-treated mice the tumour invasive front was often extensively intercalated with the surrounding normal exocrine tissue. The graph on the left indicates the average fraction of encapsulated (IT), microinvasive (IC1) and highly invasive (IC2) carcinomas per mice in each experimental condition ( n = 8). We assessed the statistical significance of the different relative representation of each tumour histotype in drug-treated versus mock control mice; * p = 0.003; ** p = 0.007; § p = 6.1E−05; §§ p = 1.7E−06. Scale bars, throughout the entire figure: 100 µm (20 µm in magnified insets in panel C ).
    2004 Osmotic Minipumps, supplied by Durect Corporation, used in various techniques. Bioz Stars score: 86/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Durect Corporation 2ml2 osmotic minipumps
    Uncl-Sema3E inhibits angiogenesis and tumour growth in the RIP-Tag2 model <t>Alzet</t> <t>minipumps</t> were implanted in 12-week-old RT2 mice to release locally in the pancreas Uncl-Sema3E ( n = 14) or mock saline ( n = 12) over a period of 2 weeks; other mice were treated orally with sunitinib ( n = 8) for the same period (see Materials and Methods Section for details); two independent experiments were performed, yielding consistent results. Average global tumour burden per mice in different experimental conditions. Vessel area in treated tumours was estimated based on CD31 staining ( n = 6, per each experimental group). Vessel coverage with mural cells was assessed by double staining for NG2 and CD31 ( n = 6). Hypoxic areas in islet tumours were revealed by staining with pimonidazol (PIMO, see Materials and Methods Section; n = 6); the same sections were costained with the endothelial cell marker Endoglin-CD105, and with DAPI. Primary RT2 tumour invasiveness was assessed by morphometric analysis of H E stained sections, as previously reported (Pàez-Ribes et al, 2009 ). Representative images of the different groups are shown on the right. Uncl-Sema3E treated tumours often displayed a non-invasive phenotype: i.e. the tumour was ‘encapsulated’ and well separated from the surrounding exocrine tissue (delimited by a dotted line). In contrast, in sunitinib-treated mice the tumour invasive front was often extensively intercalated with the surrounding normal exocrine tissue. The graph on the left indicates the average fraction of encapsulated (IT), microinvasive (IC1) and highly invasive (IC2) carcinomas per mice in each experimental condition ( n = 8). We assessed the statistical significance of the different relative representation of each tumour histotype in drug-treated versus mock control mice; * p = 0.003; ** p = 0.007; § p = 6.1E−05; §§ p = 1.7E−06. Scale bars, throughout the entire figure: 100 µm (20 µm in magnified insets in panel C ).
    2ml2 Osmotic Minipumps, supplied by Durect Corporation, used in various techniques. Bioz Stars score: 88/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Durect Corporation 2002 osmotic minipump
    Uncl-Sema3E inhibits angiogenesis and tumour growth in the RIP-Tag2 model <t>Alzet</t> <t>minipumps</t> were implanted in 12-week-old RT2 mice to release locally in the pancreas Uncl-Sema3E ( n = 14) or mock saline ( n = 12) over a period of 2 weeks; other mice were treated orally with sunitinib ( n = 8) for the same period (see Materials and Methods Section for details); two independent experiments were performed, yielding consistent results. Average global tumour burden per mice in different experimental conditions. Vessel area in treated tumours was estimated based on CD31 staining ( n = 6, per each experimental group). Vessel coverage with mural cells was assessed by double staining for NG2 and CD31 ( n = 6). Hypoxic areas in islet tumours were revealed by staining with pimonidazol (PIMO, see Materials and Methods Section; n = 6); the same sections were costained with the endothelial cell marker Endoglin-CD105, and with DAPI. Primary RT2 tumour invasiveness was assessed by morphometric analysis of H E stained sections, as previously reported (Pàez-Ribes et al, 2009 ). Representative images of the different groups are shown on the right. Uncl-Sema3E treated tumours often displayed a non-invasive phenotype: i.e. the tumour was ‘encapsulated’ and well separated from the surrounding exocrine tissue (delimited by a dotted line). In contrast, in sunitinib-treated mice the tumour invasive front was often extensively intercalated with the surrounding normal exocrine tissue. The graph on the left indicates the average fraction of encapsulated (IT), microinvasive (IC1) and highly invasive (IC2) carcinomas per mice in each experimental condition ( n = 8). We assessed the statistical significance of the different relative representation of each tumour histotype in drug-treated versus mock control mice; * p = 0.003; ** p = 0.007; § p = 6.1E−05; §§ p = 1.7E−06. Scale bars, throughout the entire figure: 100 µm (20 µm in magnified insets in panel C ).
    2002 Osmotic Minipump, supplied by Durect Corporation, used in various techniques. Bioz Stars score: 85/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Durect Corporation implantable alzet osmotic minipumps
    Blockade of αv integrins by a small molecule (CWHM 12) attenuates established skeletal muscle and cardiac fibrosis, and αv integrins represent a tractable therapeutic target in human muscle fibrosis. a Dosing regime in the therapeutic skeletal muscle fibrosis model. <t>Alzet</t> osmotic <t>minipumps</t> containing CWHM 12 or CWHM 96 (control) were inserted ten days after intramuscular CTX injection. Tissues were harvested at day 21 following CTX injection. b Representative images of picrosirius red stained sections from control- and CWHM 12-treated mice. Scale bar 25 μm. c Digital image analysis quantification of collagen (picrosirius red staining) ( n = 10). d Dosing regime in the therapeutic cardiac fibrosis model. Seven days following commencement of AngII treatment, Alzet osmotic minipumps containing CWHM 12 or CWHM 96 (control) were inserted. Tissues were harvested at day 14 after commencement of AngII treatment. e Representative images of picrosirius red stained sections from control- and CWHM 12-treated mice. Scale bars 1 mm in whole heart sections, 70 μm for magnified fields. f Digital image analysis quantification of collagen (picrosirius red staining) ( n = 11). g , h Flow cytometric analysis of PDGFRβ and αv integrin expression on PDGFRβ + cells from human skeletal muscle ( g ) and heart ( h ). i , j TGFβ activation by control- or CWHM 12-treated PDGRβ + cells isolated from human skeletal muscle ( i ) and cardiac muscle ( j ) ( n = 4). TGFβ activation was assessed alone, in the presence of TGFβ-blocking antibody (clone 1D11, 40 μg ml −1 ) (anti-TGFβ), and in the presence of recombinant human TGFβ1 (rhTGFβ) (300 pg ml −1 ). Data are expressed as mean ± SEM. * P
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    Durect Corporation model 1002 osmotic minipump
    The effects of RhoA/ROCK pathway suppression or AT1 receptor inhibition on VSMC migration in Ang II-infused mice. Ang II was subcutaneously infused at a rate of 1000 ng/kg/min using the Alzet <t>Model</t> 1002 osmotic minipump. The group infused with normal saline was used as the control. The right common carotid arteries of mice were injected and incubated with 1 ×10 10 pfu virus solution specific for RhoA knockdown for 20 min. The AT1 receptor inhibitor Irbesartan (50 mg/kg/d) and ROCK inhibitor fasudil (30 mg/kg/d) were added to drinking water for consumption by mice. ( A ) Masson’s trichrome staining of carotid arteries 14 days after Ang II infusion. Masson’s trichrome staining shows the collagen fibers (blue) and muscle fibers (red). We marked the location of the intima, media, and adventitia. Histogram shows the ratio of muscle fiber area in the intima. The scar bar is 50 μm. ** p
    Model 1002 Osmotic Minipump, supplied by Durect Corporation, used in various techniques. Bioz Stars score: 89/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    T cell survival is mediated by DC-secreted factors and IL-6. (A) Expression of GM-CSFR-α, GM-CSFR-β (AIC2B), and IL-2Rβ was determined by quantitative RT-PCR using cDNA of cells indicated. Naive CD62L + CD4 + T cells (lane 1) and splenic CD11c + DC (lane 5) were purified by flow cytometry (purity > 98%). Th1 (lane 2), Th2 (lane 3), and Th17 (lane 4) polarized CD4 + T cells. (B and C) DC of GM-CSF −/− or GM-CSF +/+ mice were cultured with purified CD4 + T cells from DO11.10/GM-CSF −/− or DO11.10/GM-CSF +/+ mice, respectively, in the presence of titrating amounts of OVA 323-339. (B) IL-6 levels in the culture supernatant at day 3 were determined by ELISA. (C) Proliferation measured by 3 H-Thymidine incorporation in the absence and in the presence of 20 ng/ml rIL-6. (D) Splenocytes of DO11.10/GM-CSF +/+ or GM-CSF −/− mice were cultured with 1 μM OVA 323-339 in the absence and presence of 20ng/ml rIL-6 for 3 d before staining of cells with KJ1-26 + mAb, Annexin V (AV), and PI and analysis by flow cytometry. Dot plot gated on KJ1-26 + cells shows early apoptotic cells (PI − AV + ), late apoptotic (PI + AV + ), and dead cells (PI + AV − ). (E) GM-CSF–deficient cultures were supplemented with rIL-1, rIL-2, or rIL-6. Proliferation was measured by 3 H-Thymidine incorporation after 3 d of culture. Proliferation index was calculated as described in Materials and methods. (F) CD4 + T cells purified from DO11.10/GM-CSF +/+ or DO11.10/GM-CSF −/− mice were injected i.v. into GM-CSF +/+ or GM-CSF −/− mice, respectively. 2 d later, groups of KO and WT mice were implanted with osmotic minipumps containing hIL-6 or were sham operated. Subsequently, mice were immunized with 200 μg OVA 323-339 peptide emulsified in CFA. After 7 d, draining LN and spleen cells were analyzed by flow cytometry. Symbols indicate total number of splenic KJ1-26 + cells of individual mice. Horizontal lines indicate averages of groups. Shown is one representative experiment of three performed. *,

    Journal: The Journal of Experimental Medicine

    Article Title: GM-CSF mediates autoimmunity by enhancing IL-6-dependent Th17 cell development and survival

    doi: 10.1084/jem.20071119

    Figure Lengend Snippet: T cell survival is mediated by DC-secreted factors and IL-6. (A) Expression of GM-CSFR-α, GM-CSFR-β (AIC2B), and IL-2Rβ was determined by quantitative RT-PCR using cDNA of cells indicated. Naive CD62L + CD4 + T cells (lane 1) and splenic CD11c + DC (lane 5) were purified by flow cytometry (purity > 98%). Th1 (lane 2), Th2 (lane 3), and Th17 (lane 4) polarized CD4 + T cells. (B and C) DC of GM-CSF −/− or GM-CSF +/+ mice were cultured with purified CD4 + T cells from DO11.10/GM-CSF −/− or DO11.10/GM-CSF +/+ mice, respectively, in the presence of titrating amounts of OVA 323-339. (B) IL-6 levels in the culture supernatant at day 3 were determined by ELISA. (C) Proliferation measured by 3 H-Thymidine incorporation in the absence and in the presence of 20 ng/ml rIL-6. (D) Splenocytes of DO11.10/GM-CSF +/+ or GM-CSF −/− mice were cultured with 1 μM OVA 323-339 in the absence and presence of 20ng/ml rIL-6 for 3 d before staining of cells with KJ1-26 + mAb, Annexin V (AV), and PI and analysis by flow cytometry. Dot plot gated on KJ1-26 + cells shows early apoptotic cells (PI − AV + ), late apoptotic (PI + AV + ), and dead cells (PI + AV − ). (E) GM-CSF–deficient cultures were supplemented with rIL-1, rIL-2, or rIL-6. Proliferation was measured by 3 H-Thymidine incorporation after 3 d of culture. Proliferation index was calculated as described in Materials and methods. (F) CD4 + T cells purified from DO11.10/GM-CSF +/+ or DO11.10/GM-CSF −/− mice were injected i.v. into GM-CSF +/+ or GM-CSF −/− mice, respectively. 2 d later, groups of KO and WT mice were implanted with osmotic minipumps containing hIL-6 or were sham operated. Subsequently, mice were immunized with 200 μg OVA 323-339 peptide emulsified in CFA. After 7 d, draining LN and spleen cells were analyzed by flow cytometry. Symbols indicate total number of splenic KJ1-26 + cells of individual mice. Horizontal lines indicate averages of groups. Shown is one representative experiment of three performed. *,

    Article Snippet: After 1–2 d, osmotic minipumps (Alzet 2001; DURECT Corporation) were filled with 26 μg (3 μg/d delivery) rec hIL-6 and s.c. implanted in mice.

    Techniques: Expressing, Quantitative RT-PCR, Purification, Flow Cytometry, Cytometry, Mouse Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Staining, Injection

    Blockade of αv integrins by a novel small molecule (CWHM 12) attenuates liver and lung fibrosis ( a ) Dosing regime in the prophylactic liver fibrosis model (left panel). Alzet minipumps containing CWHM 12 or vehicle were inserted, followed by CCl 4 I.P. twice weekly for 6 weeks. Picrosirius red (upper) and αSMA immunohistochemistry (lower) of liver tissue from control and CWHM 12 treated mice ( n = 6 female mice per group) after chronic CCl 4 treatment. Scale bar, 200μm. ( b ) Digital image analysis of picrosirius red staining. ( c ) Hydroxyproline analysis. ( d ) Digital image analysis of αSMA staining. ( e ) Dosing regime in the therapeutic liver fibrosis model (left panel). Mice were given CCl 4 I.P. twice weekly for 3 weeks, then Alzet minipumps containing either CWHM 12 or vehicle were inserted, followed by a further 3 weeks of CCl 4 I.P. twice weekly. Picrosirius red (upper) and αSMA immunohistochemistry (lower) of liver tissue from control and CWHM 12 treated mice ( n =14 female mice per group) after chronic CCl 4 treatment. Scale bar, 200μm. ( f ) Digital image analysis of picrosirius red staining. ( g ) Hydroxyproline analysis. ( h ) Digital image analysis of αSMA staining. ( i ) Dosing regime in the therapeutic lung fibrosis model. Alzet minipumps containing either CWHM 12 or vehicle were inserted 14 days after treatment with bleomycin or saline, and lungs were harvested at 28 days. ( j ) Picrosirius red staining of lung tissue from control and CWHM 12 treated mice 28 days after bleomycin instillation ( n = 15 female mice per group). Scale bar, 100μm (left panel). Hydroxyproline analysis (right panel). Data are mean ± s.e.m. * P

    Journal: Nature medicine

    Article Title: Selective ?v integrin depletion identifies a core, targetable molecular pathway that regulates fibrosis across solid organs

    doi: 10.1038/nm.3282

    Figure Lengend Snippet: Blockade of αv integrins by a novel small molecule (CWHM 12) attenuates liver and lung fibrosis ( a ) Dosing regime in the prophylactic liver fibrosis model (left panel). Alzet minipumps containing CWHM 12 or vehicle were inserted, followed by CCl 4 I.P. twice weekly for 6 weeks. Picrosirius red (upper) and αSMA immunohistochemistry (lower) of liver tissue from control and CWHM 12 treated mice ( n = 6 female mice per group) after chronic CCl 4 treatment. Scale bar, 200μm. ( b ) Digital image analysis of picrosirius red staining. ( c ) Hydroxyproline analysis. ( d ) Digital image analysis of αSMA staining. ( e ) Dosing regime in the therapeutic liver fibrosis model (left panel). Mice were given CCl 4 I.P. twice weekly for 3 weeks, then Alzet minipumps containing either CWHM 12 or vehicle were inserted, followed by a further 3 weeks of CCl 4 I.P. twice weekly. Picrosirius red (upper) and αSMA immunohistochemistry (lower) of liver tissue from control and CWHM 12 treated mice ( n =14 female mice per group) after chronic CCl 4 treatment. Scale bar, 200μm. ( f ) Digital image analysis of picrosirius red staining. ( g ) Hydroxyproline analysis. ( h ) Digital image analysis of αSMA staining. ( i ) Dosing regime in the therapeutic lung fibrosis model. Alzet minipumps containing either CWHM 12 or vehicle were inserted 14 days after treatment with bleomycin or saline, and lungs were harvested at 28 days. ( j ) Picrosirius red staining of lung tissue from control and CWHM 12 treated mice 28 days after bleomycin instillation ( n = 15 female mice per group). Scale bar, 100μm (left panel). Hydroxyproline analysis (right panel). Data are mean ± s.e.m. * P

    Article Snippet: We initially examined the potential of CWHM 12 to prevent liver fibrosis by inserting Alzet osmotic minipumps containing either CWHM 12 or vehicle control into mice, followed by CCl4 injections twice weekly for 6 weeks ( ).

    Techniques: Immunohistochemistry, Mouse Assay, Staining

    HSVLatEnk-infected polyarthritic rats exhibited reduced thermal hyperalgesia and improved spontaneous locomotor activity. Response (paw withdrawal) latencies ( A ) of controls ( n = 9) and HSVLatEnk-infected ( n = 15) polyarthritic rats to radiant heating (intensity 7; Ugo Basile) were measured 3 weeks after infection. Rearings ( B ) and horizontal locomotor activity ( C ) of control (sham- or HSVLatβ-gal-infected) ( n = 12) and HSVLatEnk-infected ( n = 15) polyarthritic rats in a red-lighted open field were video monitored and assessed every minute during a 7 min period. Animals were then implanted subcutaneously for 3 d with an Alzet osmotic minipump delivering 3 mg · kg −1 · d −1 of either naloxone (○) or naloxone methiodide (▵) ( antago ), and thermal hyperalgesia and locomotor activity were assessed. Performances of normal healthy rats are indicated by the horizontal dashed line and gray band in the three behavioral tests (mean ± SEM; n = 6–7). * p

    Journal: The Journal of Neuroscience

    Article Title: Therapeutic Efficacy in Experimental Polyarthritis of Viral-Driven Enkephalin Overproduction in Sensory Neurons

    doi: 10.1523/JNEUROSCI.21-20-07881.2001

    Figure Lengend Snippet: HSVLatEnk-infected polyarthritic rats exhibited reduced thermal hyperalgesia and improved spontaneous locomotor activity. Response (paw withdrawal) latencies ( A ) of controls ( n = 9) and HSVLatEnk-infected ( n = 15) polyarthritic rats to radiant heating (intensity 7; Ugo Basile) were measured 3 weeks after infection. Rearings ( B ) and horizontal locomotor activity ( C ) of control (sham- or HSVLatβ-gal-infected) ( n = 12) and HSVLatEnk-infected ( n = 15) polyarthritic rats in a red-lighted open field were video monitored and assessed every minute during a 7 min period. Animals were then implanted subcutaneously for 3 d with an Alzet osmotic minipump delivering 3 mg · kg −1 · d −1 of either naloxone (○) or naloxone methiodide (▵) ( antago ), and thermal hyperalgesia and locomotor activity were assessed. Performances of normal healthy rats are indicated by the horizontal dashed line and gray band in the three behavioral tests (mean ± SEM; n = 6–7). * p

    Article Snippet: The skin was incised at the level of the scapula, an Alzet osmotic minipump (delivery rate of 1 μl/hr; 2001 model; AlzaScientific Products, Palo Alto, CA) was implanted subcutaneously, and the incision was then sutured.

    Techniques: Infection, Activity Assay

    EC-SOD KO mice are sensitized to angiotensin II–induced proteinuric renal injury. Mice undergo unilateral nephrectomy and are then treated with 1.5 mg/kg per day of angiotensin II via an osmotic minipump and euthanized after 4 weeks. (A–G)

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: Extracellular Superoxide Dismutase Protects against Proteinuric Kidney Disease

    doi: 10.1681/ASN.2014060613

    Figure Lengend Snippet: EC-SOD KO mice are sensitized to angiotensin II–induced proteinuric renal injury. Mice undergo unilateral nephrectomy and are then treated with 1.5 mg/kg per day of angiotensin II via an osmotic minipump and euthanized after 4 weeks. (A–G)

    Article Snippet: On day 0, a subcutaneous osmotic minipump (model 2004; Alzet, Cupertino, CA) was implanted to deliver a constant infusion of angiotensin II (1.5 mg/kg per day; Sigma-Aldrich).

    Techniques: Mouse Assay

    Angiotensin II-induced hypertension is attenuated in Pkd 2 smKO mice. ( A ) Telemetric blood pressure time course showing the development of angiotensin II-induced hypertension in Pkd2 fl/fl (n = 6) and Pkd 2 smKO mice (n = 9). Osmotic minipumps containing either saline or angiotensin II were implanted one day prior to day 0. * indicates p

    Journal: eLife

    Article Title: Arterial smooth muscle cell PKD2 (TRPP1) channels regulate systemic blood pressure

    doi: 10.7554/eLife.42628

    Figure Lengend Snippet: Angiotensin II-induced hypertension is attenuated in Pkd 2 smKO mice. ( A ) Telemetric blood pressure time course showing the development of angiotensin II-induced hypertension in Pkd2 fl/fl (n = 6) and Pkd 2 smKO mice (n = 9). Osmotic minipumps containing either saline or angiotensin II were implanted one day prior to day 0. * indicates p

    Article Snippet: Angiotensin II (1.5 ng/g/min) and saline (0.9 NaCl) were infused in mice using subcutaneous osmotic minipumps (Alzet).

    Techniques: Mouse Assay

    Nicotine fails to enhance LLC tumor growth in vivo. C57BL/6J mice were injected s.c. with 1.5 × 10 6 LLC cells in both flanks. Once tumors formed, s.c. osmotic minipumps were implanted on day 0 to release 24 mg/kg nicotine daily for a total of 7 days. The left and right flank tumor volumes (length × width × height) were compared with the respective baseline tumor volumes to calculate fold change; the fold change values were averaged for each mouse. A linear mixed-model analysis revealed a significant effect of time [ F (4,39) = 25.747, P

    Journal: The Journal of Pharmacology and Experimental Therapeutics

    Article Title:

    doi: 10.1124/jpet.117.243972

    Figure Lengend Snippet: Nicotine fails to enhance LLC tumor growth in vivo. C57BL/6J mice were injected s.c. with 1.5 × 10 6 LLC cells in both flanks. Once tumors formed, s.c. osmotic minipumps were implanted on day 0 to release 24 mg/kg nicotine daily for a total of 7 days. The left and right flank tumor volumes (length × width × height) were compared with the respective baseline tumor volumes to calculate fold change; the fold change values were averaged for each mouse. A linear mixed-model analysis revealed a significant effect of time [ F (4,39) = 25.747, P

    Article Snippet: Nicotine at doses of 6, 12, or 24 mg/kg per day was also administered chronically via 7-day s.c. osmotic minipumps (model 1007D; Alzet, Cupertino, CA), which were implanted 2 days prior to paclitaxel treatment ( ).

    Techniques: In Vivo, Mouse Assay, Injection

    Effects of AT 1 R antagonist on blood pressure, sympathetic vasomotor activity, ROS level and expressions of AT 1 R, gp91phox or SOD2 protein in RVLM of young offspring exposed to maternal HFD or ND. a tissue ROS level; b SBP; c sympathetic vasomotor activity; and representative gels and densitometric analysis of results from Western blot showing changes in protein expression of ( d ) AT 1 R, ( e ) gp91 phox , or ( f ) SOD2 in RVLM of ND or HFD offspring, alone or with additional treatment with an AT1R antagonist, losartan (3 μg·μL − 1 ·h − 1 ), microinfused via a minipump into the cistern magna at age of 8 weeks for 4 weeks. Analysis was performed on tissues collected bilaterally from individual RVLM at age of 12 weeks. Data on protein expression were normalized to the average ND control value, which is set to 1.0, and are presented as mean ± SEM ( n = 6 in all groups). * P

    Journal: Journal of Biomedical Science

    Article Title: Anomalous AMPK-regulated angiotensin AT1R expression and SIRT1-mediated mitochondrial biogenesis at RVLM in hypertension programming of offspring to maternal high fructose exposure

    doi: 10.1186/s12929-020-00660-z

    Figure Lengend Snippet: Effects of AT 1 R antagonist on blood pressure, sympathetic vasomotor activity, ROS level and expressions of AT 1 R, gp91phox or SOD2 protein in RVLM of young offspring exposed to maternal HFD or ND. a tissue ROS level; b SBP; c sympathetic vasomotor activity; and representative gels and densitometric analysis of results from Western blot showing changes in protein expression of ( d ) AT 1 R, ( e ) gp91 phox , or ( f ) SOD2 in RVLM of ND or HFD offspring, alone or with additional treatment with an AT1R antagonist, losartan (3 μg·μL − 1 ·h − 1 ), microinfused via a minipump into the cistern magna at age of 8 weeks for 4 weeks. Analysis was performed on tissues collected bilaterally from individual RVLM at age of 12 weeks. Data on protein expression were normalized to the average ND control value, which is set to 1.0, and are presented as mean ± SEM ( n = 6 in all groups). * P

    Article Snippet: The outer end of the catheter was connected to a micro-osmotic minipump (Alzet 1007D; Durect Co., Cupertino, CA), which was placed under the skin in the neck region.

    Techniques: Activity Assay, Western Blot, Expressing

    Recovery of weight bearing after NgR1 treatment of chronically spinal cord contused rats A , Spontaneous improvement of open field locomotion (BBB score) as a function of time after thoracic spinal cord contusion injury for all rats prior to randomization to IgG or NgR1 treatment. Mean ± sem, n = 64. B , Schematic of experiment. Two weeks after the i.c.v. cannula implantation (12 weeks post-contusion injury), rats were assigned to one of two treatment groups. The PBS minipumps were replaced with new osmotic minipumps filled with 2.25 mg AA-NgR(310)ecto-Fc (0.29 mg/kg/day) or 2.25 mg rat IgG in 2 ml PBS. The duration of treatment was 12 weeks. A new osmotic mimipump filled with same amount of AA-NgR(310)ecto-Fc or rat IgG replaced each depleted pump every 4 weeks. C , Examples of a control rat without weight support at the end of the treatment period and two of the seven AA-NgR(310)ecto-Fc treated rats that regained weight support. D , The increase in the percentage of rats showing body weight support with at least one hindlimb as a function of time during therapy is reported. P =0.022 by repeated measures ANOVA for the effect of NgR1 versus IgG treatment, and *, P

    Journal: Annals of neurology

    Article Title: Recovery from Chronic Spinal Cord Contusion after Nogo Receptor Intervention

    doi: 10.1002/ana.22527

    Figure Lengend Snippet: Recovery of weight bearing after NgR1 treatment of chronically spinal cord contused rats A , Spontaneous improvement of open field locomotion (BBB score) as a function of time after thoracic spinal cord contusion injury for all rats prior to randomization to IgG or NgR1 treatment. Mean ± sem, n = 64. B , Schematic of experiment. Two weeks after the i.c.v. cannula implantation (12 weeks post-contusion injury), rats were assigned to one of two treatment groups. The PBS minipumps were replaced with new osmotic minipumps filled with 2.25 mg AA-NgR(310)ecto-Fc (0.29 mg/kg/day) or 2.25 mg rat IgG in 2 ml PBS. The duration of treatment was 12 weeks. A new osmotic mimipump filled with same amount of AA-NgR(310)ecto-Fc or rat IgG replaced each depleted pump every 4 weeks. C , Examples of a control rat without weight support at the end of the treatment period and two of the seven AA-NgR(310)ecto-Fc treated rats that regained weight support. D , The increase in the percentage of rats showing body weight support with at least one hindlimb as a function of time during therapy is reported. P =0.022 by repeated measures ANOVA for the effect of NgR1 versus IgG treatment, and *, P

    Article Snippet: Two weeks after cannula implantation (12 weeks post-contusion injury), the rats were reanesthetized and the minipumps were replaced with new osmotic minipumps (Alza Scientific Products) connected to the same cannula.

    Techniques:

    Uncl-Sema3E inhibits angiogenesis and tumour growth in the RIP-Tag2 model Alzet minipumps were implanted in 12-week-old RT2 mice to release locally in the pancreas Uncl-Sema3E ( n = 14) or mock saline ( n = 12) over a period of 2 weeks; other mice were treated orally with sunitinib ( n = 8) for the same period (see Materials and Methods Section for details); two independent experiments were performed, yielding consistent results. Average global tumour burden per mice in different experimental conditions. Vessel area in treated tumours was estimated based on CD31 staining ( n = 6, per each experimental group). Vessel coverage with mural cells was assessed by double staining for NG2 and CD31 ( n = 6). Hypoxic areas in islet tumours were revealed by staining with pimonidazol (PIMO, see Materials and Methods Section; n = 6); the same sections were costained with the endothelial cell marker Endoglin-CD105, and with DAPI. Primary RT2 tumour invasiveness was assessed by morphometric analysis of H E stained sections, as previously reported (Pàez-Ribes et al, 2009 ). Representative images of the different groups are shown on the right. Uncl-Sema3E treated tumours often displayed a non-invasive phenotype: i.e. the tumour was ‘encapsulated’ and well separated from the surrounding exocrine tissue (delimited by a dotted line). In contrast, in sunitinib-treated mice the tumour invasive front was often extensively intercalated with the surrounding normal exocrine tissue. The graph on the left indicates the average fraction of encapsulated (IT), microinvasive (IC1) and highly invasive (IC2) carcinomas per mice in each experimental condition ( n = 8). We assessed the statistical significance of the different relative representation of each tumour histotype in drug-treated versus mock control mice; * p = 0.003; ** p = 0.007; § p = 6.1E−05; §§ p = 1.7E−06. Scale bars, throughout the entire figure: 100 µm (20 µm in magnified insets in panel C ).

    Journal: EMBO Molecular Medicine

    Article Title: Tumour growth inhibition and anti-metastatic activity of a mutated furin-resistant Semaphorin 3E isoform

    doi: 10.1002/emmm.201100205

    Figure Lengend Snippet: Uncl-Sema3E inhibits angiogenesis and tumour growth in the RIP-Tag2 model Alzet minipumps were implanted in 12-week-old RT2 mice to release locally in the pancreas Uncl-Sema3E ( n = 14) or mock saline ( n = 12) over a period of 2 weeks; other mice were treated orally with sunitinib ( n = 8) for the same period (see Materials and Methods Section for details); two independent experiments were performed, yielding consistent results. Average global tumour burden per mice in different experimental conditions. Vessel area in treated tumours was estimated based on CD31 staining ( n = 6, per each experimental group). Vessel coverage with mural cells was assessed by double staining for NG2 and CD31 ( n = 6). Hypoxic areas in islet tumours were revealed by staining with pimonidazol (PIMO, see Materials and Methods Section; n = 6); the same sections were costained with the endothelial cell marker Endoglin-CD105, and with DAPI. Primary RT2 tumour invasiveness was assessed by morphometric analysis of H E stained sections, as previously reported (Pàez-Ribes et al, 2009 ). Representative images of the different groups are shown on the right. Uncl-Sema3E treated tumours often displayed a non-invasive phenotype: i.e. the tumour was ‘encapsulated’ and well separated from the surrounding exocrine tissue (delimited by a dotted line). In contrast, in sunitinib-treated mice the tumour invasive front was often extensively intercalated with the surrounding normal exocrine tissue. The graph on the left indicates the average fraction of encapsulated (IT), microinvasive (IC1) and highly invasive (IC2) carcinomas per mice in each experimental condition ( n = 8). We assessed the statistical significance of the different relative representation of each tumour histotype in drug-treated versus mock control mice; * p = 0.003; ** p = 0.007; § p = 6.1E−05; §§ p = 1.7E−06. Scale bars, throughout the entire figure: 100 µm (20 µm in magnified insets in panel C ).

    Article Snippet: Alzet osmotic minipumps (2002 model, Charles River Laboratories) were used to accomplish local delivery of Uncl-Sema3E in the pancreas of experimental mice between 12 and 14 weeks of age (see Supporting Information Methods for details).

    Techniques: Mouse Assay, Staining, Double Staining, Marker

    Blockade of αv integrins by a small molecule (CWHM 12) attenuates established skeletal muscle and cardiac fibrosis, and αv integrins represent a tractable therapeutic target in human muscle fibrosis. a Dosing regime in the therapeutic skeletal muscle fibrosis model. Alzet osmotic minipumps containing CWHM 12 or CWHM 96 (control) were inserted ten days after intramuscular CTX injection. Tissues were harvested at day 21 following CTX injection. b Representative images of picrosirius red stained sections from control- and CWHM 12-treated mice. Scale bar 25 μm. c Digital image analysis quantification of collagen (picrosirius red staining) ( n = 10). d Dosing regime in the therapeutic cardiac fibrosis model. Seven days following commencement of AngII treatment, Alzet osmotic minipumps containing CWHM 12 or CWHM 96 (control) were inserted. Tissues were harvested at day 14 after commencement of AngII treatment. e Representative images of picrosirius red stained sections from control- and CWHM 12-treated mice. Scale bars 1 mm in whole heart sections, 70 μm for magnified fields. f Digital image analysis quantification of collagen (picrosirius red staining) ( n = 11). g , h Flow cytometric analysis of PDGFRβ and αv integrin expression on PDGFRβ + cells from human skeletal muscle ( g ) and heart ( h ). i , j TGFβ activation by control- or CWHM 12-treated PDGRβ + cells isolated from human skeletal muscle ( i ) and cardiac muscle ( j ) ( n = 4). TGFβ activation was assessed alone, in the presence of TGFβ-blocking antibody (clone 1D11, 40 μg ml −1 ) (anti-TGFβ), and in the presence of recombinant human TGFβ1 (rhTGFβ) (300 pg ml −1 ). Data are expressed as mean ± SEM. * P

    Journal: Nature Communications

    Article Title: αv integrins on mesenchymal cells regulate skeletal and cardiac muscle fibrosis

    doi: 10.1038/s41467-017-01097-z

    Figure Lengend Snippet: Blockade of αv integrins by a small molecule (CWHM 12) attenuates established skeletal muscle and cardiac fibrosis, and αv integrins represent a tractable therapeutic target in human muscle fibrosis. a Dosing regime in the therapeutic skeletal muscle fibrosis model. Alzet osmotic minipumps containing CWHM 12 or CWHM 96 (control) were inserted ten days after intramuscular CTX injection. Tissues were harvested at day 21 following CTX injection. b Representative images of picrosirius red stained sections from control- and CWHM 12-treated mice. Scale bar 25 μm. c Digital image analysis quantification of collagen (picrosirius red staining) ( n = 10). d Dosing regime in the therapeutic cardiac fibrosis model. Seven days following commencement of AngII treatment, Alzet osmotic minipumps containing CWHM 12 or CWHM 96 (control) were inserted. Tissues were harvested at day 14 after commencement of AngII treatment. e Representative images of picrosirius red stained sections from control- and CWHM 12-treated mice. Scale bars 1 mm in whole heart sections, 70 μm for magnified fields. f Digital image analysis quantification of collagen (picrosirius red staining) ( n = 11). g , h Flow cytometric analysis of PDGFRβ and αv integrin expression on PDGFRβ + cells from human skeletal muscle ( g ) and heart ( h ). i , j TGFβ activation by control- or CWHM 12-treated PDGRβ + cells isolated from human skeletal muscle ( i ) and cardiac muscle ( j ) ( n = 4). TGFβ activation was assessed alone, in the presence of TGFβ-blocking antibody (clone 1D11, 40 μg ml −1 ) (anti-TGFβ), and in the presence of recombinant human TGFβ1 (rhTGFβ) (300 pg ml −1 ). Data are expressed as mean ± SEM. * P

    Article Snippet: Drug or vehicle (50% DMSO) were delivered by implantable Alzet osmotic minipumps (Durect).

    Techniques: Injection, Staining, Mouse Assay, Flow Cytometry, Expressing, Activation Assay, Isolation, Blocking Assay, Recombinant

    The effects of RhoA/ROCK pathway suppression or AT1 receptor inhibition on VSMC migration in Ang II-infused mice. Ang II was subcutaneously infused at a rate of 1000 ng/kg/min using the Alzet Model 1002 osmotic minipump. The group infused with normal saline was used as the control. The right common carotid arteries of mice were injected and incubated with 1 ×10 10 pfu virus solution specific for RhoA knockdown for 20 min. The AT1 receptor inhibitor Irbesartan (50 mg/kg/d) and ROCK inhibitor fasudil (30 mg/kg/d) were added to drinking water for consumption by mice. ( A ) Masson’s trichrome staining of carotid arteries 14 days after Ang II infusion. Masson’s trichrome staining shows the collagen fibers (blue) and muscle fibers (red). We marked the location of the intima, media, and adventitia. Histogram shows the ratio of muscle fiber area in the intima. The scar bar is 50 μm. ** p

    Journal: International Journal of Molecular Sciences

    Article Title: RhoA/ROCK Pathway Activation is Regulated by AT1 Receptor and Participates in Smooth Muscle Migration and Dedifferentiation via Promoting Actin Cytoskeleton Polymerization

    doi: 10.3390/ijms21155398

    Figure Lengend Snippet: The effects of RhoA/ROCK pathway suppression or AT1 receptor inhibition on VSMC migration in Ang II-infused mice. Ang II was subcutaneously infused at a rate of 1000 ng/kg/min using the Alzet Model 1002 osmotic minipump. The group infused with normal saline was used as the control. The right common carotid arteries of mice were injected and incubated with 1 ×10 10 pfu virus solution specific for RhoA knockdown for 20 min. The AT1 receptor inhibitor Irbesartan (50 mg/kg/d) and ROCK inhibitor fasudil (30 mg/kg/d) were added to drinking water for consumption by mice. ( A ) Masson’s trichrome staining of carotid arteries 14 days after Ang II infusion. Masson’s trichrome staining shows the collagen fibers (blue) and muscle fibers (red). We marked the location of the intima, media, and adventitia. Histogram shows the ratio of muscle fiber area in the intima. The scar bar is 50 μm. ** p

    Article Snippet: For Ang II infusion, mice were implanted with an Alzet Model 1002 osmotic minipump (Alzet Corp) for subcutaneous infusion of Ang II at a rate of 1000 ng/kg/min [ ].

    Techniques: Inhibition, Migration, Mouse Assay, Injection, Incubation, Staining