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  • 99
    Thermo Fisher 100 pmol random hexamer
    100 Pmol Random Hexamer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore k252a
    K252a, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1431 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Santa Cruz Biotechnology pmols
    Pmols, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 90 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Tocris ly341495
    Effect of an AMPA receptor antagonist on the antidepressant effect of <t>LY341495</t> or ketamine in the FST. Effect of systemic administration of NBQX. (a) LY341495 (1 mg/kg i.p.) or (b) ketamine (30 mg/kg i.p.) was administered 30 min
    Ly341495, supplied by Tocris, used in various techniques. Bioz Stars score: 98/100, based on 427 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    glp 1  (Abcam)
    99
    Abcam glp 1
    <t>GLP-1</t> does not acutely enter cerebral spinal fluid (CSF) from circulation but activates PKA activity in cerebral microvessels. Each rat received a systemic infusion of either saline or GLP-1 (30 pmol/kg/min) for 10 min, GLP-1 concentrations in CSF and plasma were determined and cerebral microvessels were isolated. (A) GLP-1 concentrations in CSF and plasma ( n = 4; * p
    Glp 1, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 201 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Tocris cgp57380
    Activation of MEK1/2, ERK1/2 or MNK1/2 in RVLM sustained central cardiovascular regulation associated with experimental brain stem death . Temporal changes in mean systemic arterial pressure (MSAP), hear rate (HR) or power density of low-frequency (LF) component of SAP signals in rats that received pretreatment by microinjection bilaterally into RVLM of vehicle (Veh; aCSF or 0.2% DMSO), ERK activation inhibitor peptide II (Peptide II; ERK2 inhibitor), U0126 (MEK1/2 inhibitor) or <t>CGP57380</t> (MNK1/2 inhibitor), 30 min before local application (at arrow) of aCSF or Mev (10 nmol) to the bilateral RVLM. Values are mean ± SEM, n = 5-7 animals per experimental group. * P
    Cgp57380, supplied by Tocris, used in various techniques. Bioz Stars score: 94/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Moravek Biochemicals cpm pmol
    Activation of MEK1/2, ERK1/2 or MNK1/2 in RVLM sustained central cardiovascular regulation associated with experimental brain stem death . Temporal changes in mean systemic arterial pressure (MSAP), hear rate (HR) or power density of low-frequency (LF) component of SAP signals in rats that received pretreatment by microinjection bilaterally into RVLM of vehicle (Veh; aCSF or 0.2% DMSO), ERK activation inhibitor peptide II (Peptide II; ERK2 inhibitor), U0126 (MEK1/2 inhibitor) or <t>CGP57380</t> (MNK1/2 inhibitor), 30 min before local application (at arrow) of aCSF or Mev (10 nmol) to the bilateral RVLM. Values are mean ± SEM, n = 5-7 animals per experimental group. * P
    Cpm Pmol, supplied by Moravek Biochemicals, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Waters Corporation pmol ul 1 massprep digestion standard
    Activation of MEK1/2, ERK1/2 or MNK1/2 in RVLM sustained central cardiovascular regulation associated with experimental brain stem death . Temporal changes in mean systemic arterial pressure (MSAP), hear rate (HR) or power density of low-frequency (LF) component of SAP signals in rats that received pretreatment by microinjection bilaterally into RVLM of vehicle (Veh; aCSF or 0.2% DMSO), ERK activation inhibitor peptide II (Peptide II; ERK2 inhibitor), U0126 (MEK1/2 inhibitor) or <t>CGP57380</t> (MNK1/2 inhibitor), 30 min before local application (at arrow) of aCSF or Mev (10 nmol) to the bilateral RVLM. Values are mean ± SEM, n = 5-7 animals per experimental group. * P
    Pmol Ul 1 Massprep Digestion Standard, supplied by Waters Corporation, used in various techniques. Bioz Stars score: 86/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher pmols heavy labeled peptide standards
    Activation of MEK1/2, ERK1/2 or MNK1/2 in RVLM sustained central cardiovascular regulation associated with experimental brain stem death . Temporal changes in mean systemic arterial pressure (MSAP), hear rate (HR) or power density of low-frequency (LF) component of SAP signals in rats that received pretreatment by microinjection bilaterally into RVLM of vehicle (Veh; aCSF or 0.2% DMSO), ERK activation inhibitor peptide II (Peptide II; ERK2 inhibitor), U0126 (MEK1/2 inhibitor) or <t>CGP57380</t> (MNK1/2 inhibitor), 30 min before local application (at arrow) of aCSF or Mev (10 nmol) to the bilateral RVLM. Values are mean ± SEM, n = 5-7 animals per experimental group. * P
    Pmols Heavy Labeled Peptide Standards, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Thermo Fisher pmols heavy labelled peptide standards
    Activation of MEK1/2, ERK1/2 or MNK1/2 in RVLM sustained central cardiovascular regulation associated with experimental brain stem death . Temporal changes in mean systemic arterial pressure (MSAP), hear rate (HR) or power density of low-frequency (LF) component of SAP signals in rats that received pretreatment by microinjection bilaterally into RVLM of vehicle (Veh; aCSF or 0.2% DMSO), ERK activation inhibitor peptide II (Peptide II; ERK2 inhibitor), U0126 (MEK1/2 inhibitor) or <t>CGP57380</t> (MNK1/2 inhibitor), 30 min before local application (at arrow) of aCSF or Mev (10 nmol) to the bilateral RVLM. Values are mean ± SEM, n = 5-7 animals per experimental group. * P
    Pmols Heavy Labelled Peptide Standards, supplied by Thermo Fisher, 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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    pmols  (ATUM)
    91
    ATUM pmols
    Activation of MEK1/2, ERK1/2 or MNK1/2 in RVLM sustained central cardiovascular regulation associated with experimental brain stem death . Temporal changes in mean systemic arterial pressure (MSAP), hear rate (HR) or power density of low-frequency (LF) component of SAP signals in rats that received pretreatment by microinjection bilaterally into RVLM of vehicle (Veh; aCSF or 0.2% DMSO), ERK activation inhibitor peptide II (Peptide II; ERK2 inhibitor), U0126 (MEK1/2 inhibitor) or <t>CGP57380</t> (MNK1/2 inhibitor), 30 min before local application (at arrow) of aCSF or Mev (10 nmol) to the bilateral RVLM. Values are mean ± SEM, n = 5-7 animals per experimental group. * P
    Pmols, supplied by ATUM, used in various techniques. Bioz Stars score: 91/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher pmols
    Activation of MEK1/2, ERK1/2 or MNK1/2 in RVLM sustained central cardiovascular regulation associated with experimental brain stem death . Temporal changes in mean systemic arterial pressure (MSAP), hear rate (HR) or power density of low-frequency (LF) component of SAP signals in rats that received pretreatment by microinjection bilaterally into RVLM of vehicle (Veh; aCSF or 0.2% DMSO), ERK activation inhibitor peptide II (Peptide II; ERK2 inhibitor), U0126 (MEK1/2 inhibitor) or <t>CGP57380</t> (MNK1/2 inhibitor), 30 min before local application (at arrow) of aCSF or Mev (10 nmol) to the bilateral RVLM. Values are mean ± SEM, n = 5-7 animals per experimental group. * P
    Pmols, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 525 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    ICN Biomedicals 1000 cpm pmol
    Activation of MEK1/2, ERK1/2 or MNK1/2 in RVLM sustained central cardiovascular regulation associated with experimental brain stem death . Temporal changes in mean systemic arterial pressure (MSAP), hear rate (HR) or power density of low-frequency (LF) component of SAP signals in rats that received pretreatment by microinjection bilaterally into RVLM of vehicle (Veh; aCSF or 0.2% DMSO), ERK activation inhibitor peptide II (Peptide II; ERK2 inhibitor), U0126 (MEK1/2 inhibitor) or <t>CGP57380</t> (MNK1/2 inhibitor), 30 min before local application (at arrow) of aCSF or Mev (10 nmol) to the bilateral RVLM. Values are mean ± SEM, n = 5-7 animals per experimental group. * P
    1000 Cpm Pmol, supplied by ICN Biomedicals, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    GE Healthcare 20 pmol primer
    Activation of MEK1/2, ERK1/2 or MNK1/2 in RVLM sustained central cardiovascular regulation associated with experimental brain stem death . Temporal changes in mean systemic arterial pressure (MSAP), hear rate (HR) or power density of low-frequency (LF) component of SAP signals in rats that received pretreatment by microinjection bilaterally into RVLM of vehicle (Veh; aCSF or 0.2% DMSO), ERK activation inhibitor peptide II (Peptide II; ERK2 inhibitor), U0126 (MEK1/2 inhibitor) or <t>CGP57380</t> (MNK1/2 inhibitor), 30 min before local application (at arrow) of aCSF or Mev (10 nmol) to the bilateral RVLM. Values are mean ± SEM, n = 5-7 animals per experimental group. * P
    20 Pmol Primer, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher 25 pmol
    Activation of MEK1/2, ERK1/2 or MNK1/2 in RVLM sustained central cardiovascular regulation associated with experimental brain stem death . Temporal changes in mean systemic arterial pressure (MSAP), hear rate (HR) or power density of low-frequency (LF) component of SAP signals in rats that received pretreatment by microinjection bilaterally into RVLM of vehicle (Veh; aCSF or 0.2% DMSO), ERK activation inhibitor peptide II (Peptide II; ERK2 inhibitor), U0126 (MEK1/2 inhibitor) or <t>CGP57380</t> (MNK1/2 inhibitor), 30 min before local application (at arrow) of aCSF or Mev (10 nmol) to the bilateral RVLM. Values are mean ± SEM, n = 5-7 animals per experimental group. * P
    25 Pmol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    American Radiolabeled Chemicals Inc 400 dpm pmol american
    Activation of MEK1/2, ERK1/2 or MNK1/2 in RVLM sustained central cardiovascular regulation associated with experimental brain stem death . Temporal changes in mean systemic arterial pressure (MSAP), hear rate (HR) or power density of low-frequency (LF) component of SAP signals in rats that received pretreatment by microinjection bilaterally into RVLM of vehicle (Veh; aCSF or 0.2% DMSO), ERK activation inhibitor peptide II (Peptide II; ERK2 inhibitor), U0126 (MEK1/2 inhibitor) or <t>CGP57380</t> (MNK1/2 inhibitor), 30 min before local application (at arrow) of aCSF or Mev (10 nmol) to the bilateral RVLM. Values are mean ± SEM, n = 5-7 animals per experimental group. * P
    400 Dpm Pmol American, supplied by American Radiolabeled Chemicals Inc, used in various techniques. Bioz Stars score: 85/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    MWG-Biotech 50 pmol oligonucleotide primer solution
    Activation of MEK1/2, ERK1/2 or MNK1/2 in RVLM sustained central cardiovascular regulation associated with experimental brain stem death . Temporal changes in mean systemic arterial pressure (MSAP), hear rate (HR) or power density of low-frequency (LF) component of SAP signals in rats that received pretreatment by microinjection bilaterally into RVLM of vehicle (Veh; aCSF or 0.2% DMSO), ERK activation inhibitor peptide II (Peptide II; ERK2 inhibitor), U0126 (MEK1/2 inhibitor) or <t>CGP57380</t> (MNK1/2 inhibitor), 30 min before local application (at arrow) of aCSF or Mev (10 nmol) to the bilateral RVLM. Values are mean ± SEM, n = 5-7 animals per experimental group. * P
    50 Pmol Oligonucleotide Primer Solution, supplied by MWG-Biotech, used in various techniques. Bioz Stars score: 85/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher 7 5 pmol oligo
    Activation of MEK1/2, ERK1/2 or MNK1/2 in RVLM sustained central cardiovascular regulation associated with experimental brain stem death . Temporal changes in mean systemic arterial pressure (MSAP), hear rate (HR) or power density of low-frequency (LF) component of SAP signals in rats that received pretreatment by microinjection bilaterally into RVLM of vehicle (Veh; aCSF or 0.2% DMSO), ERK activation inhibitor peptide II (Peptide II; ERK2 inhibitor), U0126 (MEK1/2 inhibitor) or <t>CGP57380</t> (MNK1/2 inhibitor), 30 min before local application (at arrow) of aCSF or Mev (10 nmol) to the bilateral RVLM. Values are mean ± SEM, n = 5-7 animals per experimental group. * P
    7 5 Pmol Oligo, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher t4 polynucleotide kinase
    Activation of MEK1/2, ERK1/2 or MNK1/2 in RVLM sustained central cardiovascular regulation associated with experimental brain stem death . Temporal changes in mean systemic arterial pressure (MSAP), hear rate (HR) or power density of low-frequency (LF) component of SAP signals in rats that received pretreatment by microinjection bilaterally into RVLM of vehicle (Veh; aCSF or 0.2% DMSO), ERK activation inhibitor peptide II (Peptide II; ERK2 inhibitor), U0126 (MEK1/2 inhibitor) or <t>CGP57380</t> (MNK1/2 inhibitor), 30 min before local application (at arrow) of aCSF or Mev (10 nmol) to the bilateral RVLM. Values are mean ± SEM, n = 5-7 animals per experimental group. * P
    T4 Polynucleotide Kinase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 9937 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Tocris tertiapin q
    Inhibitory effect of citalopram on the firing rate of dorsal raphe (DR) neurons in wild-type (WT) and <t>tertiapin-Q</t> injected mice. (A-B) Representative firing rate histograms illustrate the inhibitory effect of citalopram (0.5–2.5mg/kg, i.p.) on DR basal activity in artificial cerebrospinal fluid (ACSF)-injected mice (WT control, i.c.v.) (A) and tertiapin-Q injected mice (WT TPN-Q, 100 pmol, i.c.v.) (B). (C) Dose-response curves for citalopram (0.5–3mg/kg, i.p.) on DR firing rate in WT, WT control, and WT TPN-Q. Each point represents the mean±SEM of n experiments (n=4–5 mice/group).
    Tertiapin Q, supplied by Tocris, used in various techniques. Bioz Stars score: 94/100, based on 126 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore cyp1a2
    Inhibitory effect of citalopram on the firing rate of dorsal raphe (DR) neurons in wild-type (WT) and <t>tertiapin-Q</t> injected mice. (A-B) Representative firing rate histograms illustrate the inhibitory effect of citalopram (0.5–2.5mg/kg, i.p.) on DR basal activity in artificial cerebrospinal fluid (ACSF)-injected mice (WT control, i.c.v.) (A) and tertiapin-Q injected mice (WT TPN-Q, 100 pmol, i.c.v.) (B). (C) Dose-response curves for citalopram (0.5–3mg/kg, i.p.) on DR firing rate in WT, WT control, and WT TPN-Q. Each point represents the mean±SEM of n experiments (n=4–5 mice/group).
    Cyp1a2, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 86 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Avanti Polar brain ceramide
    Effects of intracellular calcium chelation by BAPTA on C 2 <t>-ceramide-</t> vs. bradykinin-activated eNOS translocation. BAEC that had been incubated with or without BAPTA/AM (20 μM for 20 min) were then treated with bradykinin (BK; 1 μM for 5 min) or C 2 -ceramide (5 μM for 30 min), as indicated in the figure, and then processed and analyzed for eNOS membrane-positive immunostaining as described in the text. Each data point represents the mean ± SE derived from analysis of 100–120 cells per treatment in three independent culture preparations.
    Brain Ceramide, supplied by Avanti Polar, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    PeproTech human recombinant il 5 ril 5
    Effect of subcutaneous <t>rIL-5</t> administration on eosinophil recruitment into challenged areas of skin. (a) Guinea-pigs were challenged with several doses of DNCB on the dorsal skin. Thereafter, 10 pmol/kg rIL-5 was injected s.c. into the dorsal surface of the left ear lobe 0 or 12 h after challenge. At 24 h after challenge, lesional skin was excised and the number of eosinophils was enumerated. Recombinant <t>IL-5</t> administered 12 h after challenge induced a small but significant increase in the number of accumulating eosinophils in the dorsal skin challenged with 0·15% DNCB. (b) Guinea-pigs challenged on the right ear lobe with 0·15% DNCB were administered s.c. with several doses of rIL-5 into the left ear lobe 0 or 12 h after challenge. At 24 h after challenge, the right ear lobe was removed and the number of eosinophils was enumerated. Whereas rIL-5 (10 pmol/kg) injected 12 h after challenge remarkably enhanced eosinophil infiltration into the right ear lobe, tissue eosinophila was suppressed when rIL-5 was administered at the time of challenge (0 h). Each group consisted of at least four animals.
    Human Recombinant Il 5 Ril 5, supplied by PeproTech, used in various techniques. Bioz Stars score: 95/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Corning Life Sciences sirna 0 8 pmol lipid
    Effect of subcutaneous <t>rIL-5</t> administration on eosinophil recruitment into challenged areas of skin. (a) Guinea-pigs were challenged with several doses of DNCB on the dorsal skin. Thereafter, 10 pmol/kg rIL-5 was injected s.c. into the dorsal surface of the left ear lobe 0 or 12 h after challenge. At 24 h after challenge, lesional skin was excised and the number of eosinophils was enumerated. Recombinant <t>IL-5</t> administered 12 h after challenge induced a small but significant increase in the number of accumulating eosinophils in the dorsal skin challenged with 0·15% DNCB. (b) Guinea-pigs challenged on the right ear lobe with 0·15% DNCB were administered s.c. with several doses of rIL-5 into the left ear lobe 0 or 12 h after challenge. At 24 h after challenge, the right ear lobe was removed and the number of eosinophils was enumerated. Whereas rIL-5 (10 pmol/kg) injected 12 h after challenge remarkably enhanced eosinophil infiltration into the right ear lobe, tissue eosinophila was suppressed when rIL-5 was administered at the time of challenge (0 h). Each group consisted of at least four animals.
    Sirna 0 8 Pmol Lipid, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 86/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effect of an AMPA receptor antagonist on the antidepressant effect of LY341495 or ketamine in the FST. Effect of systemic administration of NBQX. (a) LY341495 (1 mg/kg i.p.) or (b) ketamine (30 mg/kg i.p.) was administered 30 min

    Journal: Neuropsychopharmacology

    Article Title: The Antidepressant Effects of an mGlu2/3 Receptor Antagonist and Ketamine Require AMPA Receptor Stimulation in the mPFC and Subsequent Activation of the 5-HT Neurons in the DRN

    doi: 10.1038/npp.2015.233

    Figure Lengend Snippet: Effect of an AMPA receptor antagonist on the antidepressant effect of LY341495 or ketamine in the FST. Effect of systemic administration of NBQX. (a) LY341495 (1 mg/kg i.p.) or (b) ketamine (30 mg/kg i.p.) was administered 30 min

    Article Snippet: The decrease in the immobility time induced by microinjection of LY341495 (0.03 pmol/0.1 μl/side) into the mPFC was blocked by pretreatment with PCPA (300 mg/kg i.p. twice daily for 3 days) (LY341495, F(1, 28)=18.11, p < 0.001; PCPA, F(1, 28)=16.02, p < 0.001; interaction, F(1, 28)=21.10, p < 0.001; ).

    Techniques:

    Effect of microinjection of LY341495 or ketamine on the c-Fos immunoreactivity colocalized with 5-HT neuron cells in the dorsal raphe nucleus. (a) Confocal images of c-Fos (red), 5-HT cells (green), and double (5-HT/c-Fos, colocalization) immunoreactivities

    Journal: Neuropsychopharmacology

    Article Title: The Antidepressant Effects of an mGlu2/3 Receptor Antagonist and Ketamine Require AMPA Receptor Stimulation in the mPFC and Subsequent Activation of the 5-HT Neurons in the DRN

    doi: 10.1038/npp.2015.233

    Figure Lengend Snippet: Effect of microinjection of LY341495 or ketamine on the c-Fos immunoreactivity colocalized with 5-HT neuron cells in the dorsal raphe nucleus. (a) Confocal images of c-Fos (red), 5-HT cells (green), and double (5-HT/c-Fos, colocalization) immunoreactivities

    Article Snippet: The decrease in the immobility time induced by microinjection of LY341495 (0.03 pmol/0.1 μl/side) into the mPFC was blocked by pretreatment with PCPA (300 mg/kg i.p. twice daily for 3 days) (LY341495, F(1, 28)=18.11, p < 0.001; PCPA, F(1, 28)=16.02, p < 0.001; interaction, F(1, 28)=21.10, p < 0.001; ).

    Techniques:

    Effect of microinjection of an AMPA receptor antagonist on LY341495- or ketamine-induced c-Fos immunoreactivity colocalized with 5-HT neuron cells in the dorsal raphe nucleus. (a, b) Confocal images of c-Fos (red), 5-HT cells (green), and double (5-HT/c-Fos,

    Journal: Neuropsychopharmacology

    Article Title: The Antidepressant Effects of an mGlu2/3 Receptor Antagonist and Ketamine Require AMPA Receptor Stimulation in the mPFC and Subsequent Activation of the 5-HT Neurons in the DRN

    doi: 10.1038/npp.2015.233

    Figure Lengend Snippet: Effect of microinjection of an AMPA receptor antagonist on LY341495- or ketamine-induced c-Fos immunoreactivity colocalized with 5-HT neuron cells in the dorsal raphe nucleus. (a, b) Confocal images of c-Fos (red), 5-HT cells (green), and double (5-HT/c-Fos,

    Article Snippet: The decrease in the immobility time induced by microinjection of LY341495 (0.03 pmol/0.1 μl/side) into the mPFC was blocked by pretreatment with PCPA (300 mg/kg i.p. twice daily for 3 days) (LY341495, F(1, 28)=18.11, p < 0.001; PCPA, F(1, 28)=16.02, p < 0.001; interaction, F(1, 28)=21.10, p < 0.001; ).

    Techniques:

    Effect of 5-HT depletion on the antidepressant effect of LY341495, ketamine, or paroxetine in the FST. Effect of LY341495 (a), ketamine (b), or paroxetine (c). LY341495 (0.1, 0.3, and 1 mg/kg i.p.), ketamine (3, 10, and 30 mg/kg i.p.),

    Journal: Neuropsychopharmacology

    Article Title: The Antidepressant Effects of an mGlu2/3 Receptor Antagonist and Ketamine Require AMPA Receptor Stimulation in the mPFC and Subsequent Activation of the 5-HT Neurons in the DRN

    doi: 10.1038/npp.2015.233

    Figure Lengend Snippet: Effect of 5-HT depletion on the antidepressant effect of LY341495, ketamine, or paroxetine in the FST. Effect of LY341495 (a), ketamine (b), or paroxetine (c). LY341495 (0.1, 0.3, and 1 mg/kg i.p.), ketamine (3, 10, and 30 mg/kg i.p.),

    Article Snippet: The decrease in the immobility time induced by microinjection of LY341495 (0.03 pmol/0.1 μl/side) into the mPFC was blocked by pretreatment with PCPA (300 mg/kg i.p. twice daily for 3 days) (LY341495, F(1, 28)=18.11, p < 0.001; PCPA, F(1, 28)=16.02, p < 0.001; interaction, F(1, 28)=21.10, p < 0.001; ).

    Techniques:

    Effect of microinjection of LY341495 or ketamine into the mPFC in the FST. Effect of microinjection of LY341495 (a) or ketamine (b). LY341495 (0.003, and 0.03 pmol/side) or ketamine (0.3 and 3 nmol/side) was administered into the mPFC

    Journal: Neuropsychopharmacology

    Article Title: The Antidepressant Effects of an mGlu2/3 Receptor Antagonist and Ketamine Require AMPA Receptor Stimulation in the mPFC and Subsequent Activation of the 5-HT Neurons in the DRN

    doi: 10.1038/npp.2015.233

    Figure Lengend Snippet: Effect of microinjection of LY341495 or ketamine into the mPFC in the FST. Effect of microinjection of LY341495 (a) or ketamine (b). LY341495 (0.003, and 0.03 pmol/side) or ketamine (0.3 and 3 nmol/side) was administered into the mPFC

    Article Snippet: The decrease in the immobility time induced by microinjection of LY341495 (0.03 pmol/0.1 μl/side) into the mPFC was blocked by pretreatment with PCPA (300 mg/kg i.p. twice daily for 3 days) (LY341495, F(1, 28)=18.11, p < 0.001; PCPA, F(1, 28)=16.02, p < 0.001; interaction, F(1, 28)=21.10, p < 0.001; ).

    Techniques:

    GLP-1 does not acutely enter cerebral spinal fluid (CSF) from circulation but activates PKA activity in cerebral microvessels. Each rat received a systemic infusion of either saline or GLP-1 (30 pmol/kg/min) for 10 min, GLP-1 concentrations in CSF and plasma were determined and cerebral microvessels were isolated. (A) GLP-1 concentrations in CSF and plasma ( n = 4; * p

    Journal: Frontiers in Physiology

    Article Title: Brain Endothelial Cells Regulate Glucagon-Like Peptide 1 Entry Into the Brain via a Receptor-Mediated Process

    doi: 10.3389/fphys.2020.00555

    Figure Lengend Snippet: GLP-1 does not acutely enter cerebral spinal fluid (CSF) from circulation but activates PKA activity in cerebral microvessels. Each rat received a systemic infusion of either saline or GLP-1 (30 pmol/kg/min) for 10 min, GLP-1 concentrations in CSF and plasma were determined and cerebral microvessels were isolated. (A) GLP-1 concentrations in CSF and plasma ( n = 4; * p

    Article Snippet: After 30 min of stabilization, the rat received a continuous i.v. infusion of GLP-1-FAM (30 pmol/kg/min), GLP-1(30 pmol/kg/min) or saline or an i.v. injection of liraglutide (30 pmol/kg) along with a bolus i.v. injection of glucose (0.5 g/kg).

    Techniques: Activity Assay, Isolation

    GLP-1 infusion increases muscle but not hypothalamic microvascular perfusion. Each rat received a systemic infusion of saline or GLP-1 at 30 pmol/kg/min for 120 min and MBV, MFV and MBF were determined using contrast-enhanced ultrasound in both hypothalamus (A,B,C) and muscle (D,E,F) ( n = 6, * p

    Journal: Frontiers in Physiology

    Article Title: Brain Endothelial Cells Regulate Glucagon-Like Peptide 1 Entry Into the Brain via a Receptor-Mediated Process

    doi: 10.3389/fphys.2020.00555

    Figure Lengend Snippet: GLP-1 infusion increases muscle but not hypothalamic microvascular perfusion. Each rat received a systemic infusion of saline or GLP-1 at 30 pmol/kg/min for 120 min and MBV, MFV and MBF were determined using contrast-enhanced ultrasound in both hypothalamus (A,B,C) and muscle (D,E,F) ( n = 6, * p

    Article Snippet: After 30 min of stabilization, the rat received a continuous i.v. infusion of GLP-1-FAM (30 pmol/kg/min), GLP-1(30 pmol/kg/min) or saline or an i.v. injection of liraglutide (30 pmol/kg) along with a bolus i.v. injection of glucose (0.5 g/kg).

    Techniques:

    GLP-1 infusion increases PKA activity in various brain regions in both normal and vagotomized rats. Each rat received either saline or GLP-1 (30 pmol/kg/min) infusion via a carotid artery catheter for 10 min. PKA activities in various brain regions were determined in normal rats (A,B) and vagotomized rats (D,E) . (C) Fluorogold stain of dorsal motor vagal nucleus. Lack of staining in vagotomized rats confirms surgical completeness of vagotomy. (A,D) , PKA activity. (B,E) , Quantitative analysis ( B , n = 7; * p

    Journal: Frontiers in Physiology

    Article Title: Brain Endothelial Cells Regulate Glucagon-Like Peptide 1 Entry Into the Brain via a Receptor-Mediated Process

    doi: 10.3389/fphys.2020.00555

    Figure Lengend Snippet: GLP-1 infusion increases PKA activity in various brain regions in both normal and vagotomized rats. Each rat received either saline or GLP-1 (30 pmol/kg/min) infusion via a carotid artery catheter for 10 min. PKA activities in various brain regions were determined in normal rats (A,B) and vagotomized rats (D,E) . (C) Fluorogold stain of dorsal motor vagal nucleus. Lack of staining in vagotomized rats confirms surgical completeness of vagotomy. (A,D) , PKA activity. (B,E) , Quantitative analysis ( B , n = 7; * p

    Article Snippet: After 30 min of stabilization, the rat received a continuous i.v. infusion of GLP-1-FAM (30 pmol/kg/min), GLP-1(30 pmol/kg/min) or saline or an i.v. injection of liraglutide (30 pmol/kg) along with a bolus i.v. injection of glucose (0.5 g/kg).

    Techniques: Activity Assay, Staining

    GLP-1 uptake by RBMECs is GLP-1 receptor and PKA dependent. RBMECs were pretreated with vehicle (PSS buffer, A,E,I ), Exendin 9-39 (10 nM, B,F,J ) or H89 (10 μM, C,G,K ) for 30 min. Cells were then incubated with GLP-1-FAM at 10, 100 pM or 1 nM for 3 min. DAPI was used to stain the nucleus. Images were captured using confocal microscope. Fluorescence density was analyzed using ImageJ. (D,H,L) Quantitative analysis ( n = 3, * p

    Journal: Frontiers in Physiology

    Article Title: Brain Endothelial Cells Regulate Glucagon-Like Peptide 1 Entry Into the Brain via a Receptor-Mediated Process

    doi: 10.3389/fphys.2020.00555

    Figure Lengend Snippet: GLP-1 uptake by RBMECs is GLP-1 receptor and PKA dependent. RBMECs were pretreated with vehicle (PSS buffer, A,E,I ), Exendin 9-39 (10 nM, B,F,J ) or H89 (10 μM, C,G,K ) for 30 min. Cells were then incubated with GLP-1-FAM at 10, 100 pM or 1 nM for 3 min. DAPI was used to stain the nucleus. Images were captured using confocal microscope. Fluorescence density was analyzed using ImageJ. (D,H,L) Quantitative analysis ( n = 3, * p

    Article Snippet: After 30 min of stabilization, the rat received a continuous i.v. infusion of GLP-1-FAM (30 pmol/kg/min), GLP-1(30 pmol/kg/min) or saline or an i.v. injection of liraglutide (30 pmol/kg) along with a bolus i.v. injection of glucose (0.5 g/kg).

    Techniques: Incubation, Staining, Microscopy, Fluorescence

    Expression of GLP-1 receptor in various blood vessels, RBMECs, and astrocytes. (A) Quantitative analysis of Western blot determination of GLP-1 receptor expression in cultured cells and isolated vessels ( n = 5–7, * p

    Journal: Frontiers in Physiology

    Article Title: Brain Endothelial Cells Regulate Glucagon-Like Peptide 1 Entry Into the Brain via a Receptor-Mediated Process

    doi: 10.3389/fphys.2020.00555

    Figure Lengend Snippet: Expression of GLP-1 receptor in various blood vessels, RBMECs, and astrocytes. (A) Quantitative analysis of Western blot determination of GLP-1 receptor expression in cultured cells and isolated vessels ( n = 5–7, * p

    Article Snippet: After 30 min of stabilization, the rat received a continuous i.v. infusion of GLP-1-FAM (30 pmol/kg/min), GLP-1(30 pmol/kg/min) or saline or an i.v. injection of liraglutide (30 pmol/kg) along with a bolus i.v. injection of glucose (0.5 g/kg).

    Techniques: Expressing, Western Blot, Cell Culture, Isolation

    cAMP agonist increases GLP-1 uptake by RBMECs. (A) RBMECs were incubated with GLP-1-FAM at 10 pM for 3 min with or without 10 μM forskolin. DAPI was used to stain the nucleus. Images was captured using confocal microscope. Fluorescence density was analyzed using ImageJ. (B) quantitative analysis of the images ( n = 6, * p

    Journal: Frontiers in Physiology

    Article Title: Brain Endothelial Cells Regulate Glucagon-Like Peptide 1 Entry Into the Brain via a Receptor-Mediated Process

    doi: 10.3389/fphys.2020.00555

    Figure Lengend Snippet: cAMP agonist increases GLP-1 uptake by RBMECs. (A) RBMECs were incubated with GLP-1-FAM at 10 pM for 3 min with or without 10 μM forskolin. DAPI was used to stain the nucleus. Images was captured using confocal microscope. Fluorescence density was analyzed using ImageJ. (B) quantitative analysis of the images ( n = 6, * p

    Article Snippet: After 30 min of stabilization, the rat received a continuous i.v. infusion of GLP-1-FAM (30 pmol/kg/min), GLP-1(30 pmol/kg/min) or saline or an i.v. injection of liraglutide (30 pmol/kg) along with a bolus i.v. injection of glucose (0.5 g/kg).

    Techniques: Incubation, Staining, Microscopy, Fluorescence

    GLP-1 and GLP-1 analog exendin-4 enter various brain regions via GLP-1 receptor-mediated process. (A) Protocol. Each rat received saline or exendin-(9-39) (30 nmol/kg/min) for 10 min and then dextran-TRITC (30 pmol/kg/min) + GLP-1-FAM (30 pmol/kg/min) or exendin-4-FAM (30 pmol/kg/min) for 10 min via a carotid artery catheter. Brain tissues were harvested after systemic flush with saline. (B) GLP-1-FAM uptake by the brain ( n = 7, * p

    Journal: Frontiers in Physiology

    Article Title: Brain Endothelial Cells Regulate Glucagon-Like Peptide 1 Entry Into the Brain via a Receptor-Mediated Process

    doi: 10.3389/fphys.2020.00555

    Figure Lengend Snippet: GLP-1 and GLP-1 analog exendin-4 enter various brain regions via GLP-1 receptor-mediated process. (A) Protocol. Each rat received saline or exendin-(9-39) (30 nmol/kg/min) for 10 min and then dextran-TRITC (30 pmol/kg/min) + GLP-1-FAM (30 pmol/kg/min) or exendin-4-FAM (30 pmol/kg/min) for 10 min via a carotid artery catheter. Brain tissues were harvested after systemic flush with saline. (B) GLP-1-FAM uptake by the brain ( n = 7, * p

    Article Snippet: After 30 min of stabilization, the rat received a continuous i.v. infusion of GLP-1-FAM (30 pmol/kg/min), GLP-1(30 pmol/kg/min) or saline or an i.v. injection of liraglutide (30 pmol/kg) along with a bolus i.v. injection of glucose (0.5 g/kg).

    Techniques:

    Trans-endothelial transport of GLP-1 is a GLP-1 receptor-mediated process. (A) Illustration of trans-well apparatus. (B) Time course of GLP-1-FAM transport across the RBMEC and astrocyte barrier, without ( n = 7, * p

    Journal: Frontiers in Physiology

    Article Title: Brain Endothelial Cells Regulate Glucagon-Like Peptide 1 Entry Into the Brain via a Receptor-Mediated Process

    doi: 10.3389/fphys.2020.00555

    Figure Lengend Snippet: Trans-endothelial transport of GLP-1 is a GLP-1 receptor-mediated process. (A) Illustration of trans-well apparatus. (B) Time course of GLP-1-FAM transport across the RBMEC and astrocyte barrier, without ( n = 7, * p

    Article Snippet: After 30 min of stabilization, the rat received a continuous i.v. infusion of GLP-1-FAM (30 pmol/kg/min), GLP-1(30 pmol/kg/min) or saline or an i.v. injection of liraglutide (30 pmol/kg) along with a bolus i.v. injection of glucose (0.5 g/kg).

    Techniques:

    Activation of MEK1/2, ERK1/2 or MNK1/2 in RVLM sustained central cardiovascular regulation associated with experimental brain stem death . Temporal changes in mean systemic arterial pressure (MSAP), hear rate (HR) or power density of low-frequency (LF) component of SAP signals in rats that received pretreatment by microinjection bilaterally into RVLM of vehicle (Veh; aCSF or 0.2% DMSO), ERK activation inhibitor peptide II (Peptide II; ERK2 inhibitor), U0126 (MEK1/2 inhibitor) or CGP57380 (MNK1/2 inhibitor), 30 min before local application (at arrow) of aCSF or Mev (10 nmol) to the bilateral RVLM. Values are mean ± SEM, n = 5-7 animals per experimental group. * P

    Journal: Journal of Biomedical Science

    Article Title: Extracellular signal-regulated kinase 1/2 plays a pro-life role in experimental brain stem death via MAPK signal-interacting kinase at rostral ventrolateral medulla

    doi: 10.1186/1423-0127-17-17

    Figure Lengend Snippet: Activation of MEK1/2, ERK1/2 or MNK1/2 in RVLM sustained central cardiovascular regulation associated with experimental brain stem death . Temporal changes in mean systemic arterial pressure (MSAP), hear rate (HR) or power density of low-frequency (LF) component of SAP signals in rats that received pretreatment by microinjection bilaterally into RVLM of vehicle (Veh; aCSF or 0.2% DMSO), ERK activation inhibitor peptide II (Peptide II; ERK2 inhibitor), U0126 (MEK1/2 inhibitor) or CGP57380 (MNK1/2 inhibitor), 30 min before local application (at arrow) of aCSF or Mev (10 nmol) to the bilateral RVLM. Values are mean ± SEM, n = 5-7 animals per experimental group. * P

    Article Snippet: Intriguingly, comparable results were also obtained on pretreatment with microinjection bilaterally into RVLM of CGP57380 (5 pmol), a specific cell-permeable MNK1/2 inhibitor [ , ] (fig. ); although a dose of 1 pmol was ineffective against the cardiovascular responses during the pro-life phase (maximal MSAP: 112.5 ± 5.2 versus 113.0 ± 5.8 mmHg; maximal HR: 356.8 ± 20.1 versus 354.6 ± 16.6 bpm; maximal LF power: 73.6 ± 6.7 versus 75.2 ± 7.5 mmHg2 when compared to 0.2% DMSO pretreatment; mean ± SEM, n = 4 animals).

    Techniques: Activation Assay

    Activation of MEK1/2, ERK1/2 or MNK1/2 leads to phasic upregulation of NOS I/PKG cascade in RVLM during experimental brain stem death . Illustrative gels or summary of fold changes against aCSF controls in ratio of nitric oxide synthase I (NOS I) or protein kinase G (PKG) relative to β-actin protein detected in ventrolateral medulla of rats that received ERK activation inhibitor peptide II, U0126 or CGP57380 into bilateral RVLM, 30 min before induction of experimental brain stem death. Values are mean ± SEM of triplicate analyses on samples pooled from 5-7 animals per experimental group. * P

    Journal: Journal of Biomedical Science

    Article Title: Extracellular signal-regulated kinase 1/2 plays a pro-life role in experimental brain stem death via MAPK signal-interacting kinase at rostral ventrolateral medulla

    doi: 10.1186/1423-0127-17-17

    Figure Lengend Snippet: Activation of MEK1/2, ERK1/2 or MNK1/2 leads to phasic upregulation of NOS I/PKG cascade in RVLM during experimental brain stem death . Illustrative gels or summary of fold changes against aCSF controls in ratio of nitric oxide synthase I (NOS I) or protein kinase G (PKG) relative to β-actin protein detected in ventrolateral medulla of rats that received ERK activation inhibitor peptide II, U0126 or CGP57380 into bilateral RVLM, 30 min before induction of experimental brain stem death. Values are mean ± SEM of triplicate analyses on samples pooled from 5-7 animals per experimental group. * P

    Article Snippet: Intriguingly, comparable results were also obtained on pretreatment with microinjection bilaterally into RVLM of CGP57380 (5 pmol), a specific cell-permeable MNK1/2 inhibitor [ , ] (fig. ); although a dose of 1 pmol was ineffective against the cardiovascular responses during the pro-life phase (maximal MSAP: 112.5 ± 5.2 versus 113.0 ± 5.8 mmHg; maximal HR: 356.8 ± 20.1 versus 354.6 ± 16.6 bpm; maximal LF power: 73.6 ± 6.7 versus 75.2 ± 7.5 mmHg2 when compared to 0.2% DMSO pretreatment; mean ± SEM, n = 4 animals).

    Techniques: Activation Assay

    Activation of MEK1/2, ERK1/2 or MNK1/2 did not affect NOS II/peroxynitrite signaling in RVLM during experimental brain stem death . Illustrative gels or summary of fold changes against aCSF controls in ratio of NOS II or nitrotyrosine (NT; marker for peroxynitrite) relative to β-actin protein detected in ventrolateral medulla of rats that received ERK activation inhibitor peptide II, U0126 or CGP57380 into bilateral RVLM, 30 min before induction of brain stem death. Note that NT is presented as % relative to β-actin because it is below detection limit (ND) in aCSF controls. Values are mean ± SEM of triplicate analyses on samples pooled from 5-7 animals per experimental group. * P

    Journal: Journal of Biomedical Science

    Article Title: Extracellular signal-regulated kinase 1/2 plays a pro-life role in experimental brain stem death via MAPK signal-interacting kinase at rostral ventrolateral medulla

    doi: 10.1186/1423-0127-17-17

    Figure Lengend Snippet: Activation of MEK1/2, ERK1/2 or MNK1/2 did not affect NOS II/peroxynitrite signaling in RVLM during experimental brain stem death . Illustrative gels or summary of fold changes against aCSF controls in ratio of NOS II or nitrotyrosine (NT; marker for peroxynitrite) relative to β-actin protein detected in ventrolateral medulla of rats that received ERK activation inhibitor peptide II, U0126 or CGP57380 into bilateral RVLM, 30 min before induction of brain stem death. Note that NT is presented as % relative to β-actin because it is below detection limit (ND) in aCSF controls. Values are mean ± SEM of triplicate analyses on samples pooled from 5-7 animals per experimental group. * P

    Article Snippet: Intriguingly, comparable results were also obtained on pretreatment with microinjection bilaterally into RVLM of CGP57380 (5 pmol), a specific cell-permeable MNK1/2 inhibitor [ , ] (fig. ); although a dose of 1 pmol was ineffective against the cardiovascular responses during the pro-life phase (maximal MSAP: 112.5 ± 5.2 versus 113.0 ± 5.8 mmHg; maximal HR: 356.8 ± 20.1 versus 354.6 ± 16.6 bpm; maximal LF power: 73.6 ± 6.7 versus 75.2 ± 7.5 mmHg2 when compared to 0.2% DMSO pretreatment; mean ± SEM, n = 4 animals).

    Techniques: Activation Assay, Marker

    Inhibitory effect of citalopram on the firing rate of dorsal raphe (DR) neurons in wild-type (WT) and tertiapin-Q injected mice. (A-B) Representative firing rate histograms illustrate the inhibitory effect of citalopram (0.5–2.5mg/kg, i.p.) on DR basal activity in artificial cerebrospinal fluid (ACSF)-injected mice (WT control, i.c.v.) (A) and tertiapin-Q injected mice (WT TPN-Q, 100 pmol, i.c.v.) (B). (C) Dose-response curves for citalopram (0.5–3mg/kg, i.p.) on DR firing rate in WT, WT control, and WT TPN-Q. Each point represents the mean±SEM of n experiments (n=4–5 mice/group).

    Journal: International Journal of Neuropsychopharmacology

    Article Title: Deletion of GIRK2 Subunit of GIRK Channels Alters the 5-HT1A Receptor-Mediated Signaling and Results in a Depression-Resistant Behavior

    doi: 10.1093/ijnp/pyv051

    Figure Lengend Snippet: Inhibitory effect of citalopram on the firing rate of dorsal raphe (DR) neurons in wild-type (WT) and tertiapin-Q injected mice. (A-B) Representative firing rate histograms illustrate the inhibitory effect of citalopram (0.5–2.5mg/kg, i.p.) on DR basal activity in artificial cerebrospinal fluid (ACSF)-injected mice (WT control, i.c.v.) (A) and tertiapin-Q injected mice (WT TPN-Q, 100 pmol, i.c.v.) (B). (C) Dose-response curves for citalopram (0.5–3mg/kg, i.p.) on DR firing rate in WT, WT control, and WT TPN-Q. Each point represents the mean±SEM of n experiments (n=4–5 mice/group).

    Article Snippet: Next, the effect of the high-affinity GIRK channel blocker, tertiapin-Q (100 pmol, i.c.v.) ( ; ), on the electrophysiological properties of DR neurons was assessed.

    Techniques: Injection, Mouse Assay, Activity Assay

    Effects of intracellular calcium chelation by BAPTA on C 2 -ceramide- vs. bradykinin-activated eNOS translocation. BAEC that had been incubated with or without BAPTA/AM (20 μM for 20 min) were then treated with bradykinin (BK; 1 μM for 5 min) or C 2 -ceramide (5 μM for 30 min), as indicated in the figure, and then processed and analyzed for eNOS membrane-positive immunostaining as described in the text. Each data point represents the mean ± SE derived from analysis of 100–120 cells per treatment in three independent culture preparations.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Calcium-independent activation of endothelial nitric oxide synthase by ceramide

    doi:

    Figure Lengend Snippet: Effects of intracellular calcium chelation by BAPTA on C 2 -ceramide- vs. bradykinin-activated eNOS translocation. BAEC that had been incubated with or without BAPTA/AM (20 μM for 20 min) were then treated with bradykinin (BK; 1 μM for 5 min) or C 2 -ceramide (5 μM for 30 min), as indicated in the figure, and then processed and analyzed for eNOS membrane-positive immunostaining as described in the text. Each data point represents the mean ± SE derived from analysis of 100–120 cells per treatment in three independent culture preparations.

    Article Snippet: Ceramide content was quantified within each experiment by using a standard curve established by assaying samples of authentic brain ceramide (50–400 pmol) processed and analyzed in parallel; the reference ceramide yielded an R f value of 0.25, as reported ( ).

    Techniques: Translocation Assay, Incubation, Immunostaining, Derivative Assay

    Ceramide generation by bradykinin in BAEC. BAEC were treated with bradykinin, then processed for quantitation of cellular ceramide content. ( A ) Time course of ceramide generation by bradykinin. After addition of bradykinin (10 μM), endothelial cells grown in six-well dishes were harvested at the indicated times, and cellular ceramide content was quantified as described in Methods . The relative increase over basal cellular ceramide content (which ranged from 150 to 300 pmol per well) was calculated at each time point; each data point represents the mean ± SE derived from six independent culture preparations, each performed in duplicate. ( B ) Dose dependence of ceramide generation by bradykinin. Endothelial cells were treated with the indicated concentrations of bradykinin for 3 min, then harvested and processed for ceramide content. The cellular ceramide content at each dose was normalized relative to the basal value determined for vehicle-treated cells within each experiment; each data point represents the mean ± SE derived from five independent culture preparations, each analyzed in duplicate. The single data point with the open circle shows the relative ceramide content in cells treated with bradykinin (1 μM) plus the bradykinin B 2 -receptor antagonist HOE140 (3 μM).

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Calcium-independent activation of endothelial nitric oxide synthase by ceramide

    doi:

    Figure Lengend Snippet: Ceramide generation by bradykinin in BAEC. BAEC were treated with bradykinin, then processed for quantitation of cellular ceramide content. ( A ) Time course of ceramide generation by bradykinin. After addition of bradykinin (10 μM), endothelial cells grown in six-well dishes were harvested at the indicated times, and cellular ceramide content was quantified as described in Methods . The relative increase over basal cellular ceramide content (which ranged from 150 to 300 pmol per well) was calculated at each time point; each data point represents the mean ± SE derived from six independent culture preparations, each performed in duplicate. ( B ) Dose dependence of ceramide generation by bradykinin. Endothelial cells were treated with the indicated concentrations of bradykinin for 3 min, then harvested and processed for ceramide content. The cellular ceramide content at each dose was normalized relative to the basal value determined for vehicle-treated cells within each experiment; each data point represents the mean ± SE derived from five independent culture preparations, each analyzed in duplicate. The single data point with the open circle shows the relative ceramide content in cells treated with bradykinin (1 μM) plus the bradykinin B 2 -receptor antagonist HOE140 (3 μM).

    Article Snippet: Ceramide content was quantified within each experiment by using a standard curve established by assaying samples of authentic brain ceramide (50–400 pmol) processed and analyzed in parallel; the reference ceramide yielded an R f value of 0.25, as reported ( ).

    Techniques: Quantitation Assay, Derivative Assay

    Translocation of eNOS in endothelial cells in response to C 2 -ceramide. Shown are typical photomicrographs of BAEC processed for immunofluorescence as described in the text. ( A ) A representative image of eNOS-immunolabeled endothelial cells studied under basal conditions. ( B ) A representative image after treatment of endothelial cells with C 2 -ceramide (5 μM for 30 min). Both images are shown at 250× magnification and printed with identical brightness and contrast settings.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Calcium-independent activation of endothelial nitric oxide synthase by ceramide

    doi:

    Figure Lengend Snippet: Translocation of eNOS in endothelial cells in response to C 2 -ceramide. Shown are typical photomicrographs of BAEC processed for immunofluorescence as described in the text. ( A ) A representative image of eNOS-immunolabeled endothelial cells studied under basal conditions. ( B ) A representative image after treatment of endothelial cells with C 2 -ceramide (5 μM for 30 min). Both images are shown at 250× magnification and printed with identical brightness and contrast settings.

    Article Snippet: Ceramide content was quantified within each experiment by using a standard curve established by assaying samples of authentic brain ceramide (50–400 pmol) processed and analyzed in parallel; the reference ceramide yielded an R f value of 0.25, as reported ( ).

    Techniques: Translocation Assay, Immunofluorescence, Immunolabeling

    Time course of C 2 -ceramide-induced eNOS translocation. BAEC were treated with C 2 -ceramide (5 μM) or with C 2 -dihydroceramide (5 μM) for the times indicated. The proportion of cells with eNOS immunoreactivity at the cell membrane was determined as described in Methods ); the basal value (at t = 0) was used to normalize the percentage of cells with eNOS membrane-positive staining at each subsequent time point. At each time point, 100–120 cells were scored; the data shown represent the mean ± SE derived from three independent culture preparations.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Calcium-independent activation of endothelial nitric oxide synthase by ceramide

    doi:

    Figure Lengend Snippet: Time course of C 2 -ceramide-induced eNOS translocation. BAEC were treated with C 2 -ceramide (5 μM) or with C 2 -dihydroceramide (5 μM) for the times indicated. The proportion of cells with eNOS immunoreactivity at the cell membrane was determined as described in Methods ); the basal value (at t = 0) was used to normalize the percentage of cells with eNOS membrane-positive staining at each subsequent time point. At each time point, 100–120 cells were scored; the data shown represent the mean ± SE derived from three independent culture preparations.

    Article Snippet: Ceramide content was quantified within each experiment by using a standard curve established by assaying samples of authentic brain ceramide (50–400 pmol) processed and analyzed in parallel; the reference ceramide yielded an R f value of 0.25, as reported ( ).

    Techniques: Translocation Assay, Staining, Derivative Assay

    Effects of intracellular calcium chelation by BAPTA on C 2 -ceramide- vs. bradykinin-activated NO production. BAEC were incubated with the NO indicator dye DAF-2/DA and processed for intracellular NO generation as described in the text. The integrated fluorescence intensity was measured before and 15 min after addition of bradykinin (1 μM) or C 2 -ceramide (5 μM); data shown are normalized to the fluorescence intensity measured prior to drug addition. As indicated in the figure, some cultures were incubated in the presence of the intracellular calcium chelator BAPTA/AM (20 μM for 20 min) prior to addition of bradykinin (BK) or C 2 -ceramide. For each treatment, 20–25 cells in three independent culture preparations were examined; each data point represents the mean ± SE.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Calcium-independent activation of endothelial nitric oxide synthase by ceramide

    doi:

    Figure Lengend Snippet: Effects of intracellular calcium chelation by BAPTA on C 2 -ceramide- vs. bradykinin-activated NO production. BAEC were incubated with the NO indicator dye DAF-2/DA and processed for intracellular NO generation as described in the text. The integrated fluorescence intensity was measured before and 15 min after addition of bradykinin (1 μM) or C 2 -ceramide (5 μM); data shown are normalized to the fluorescence intensity measured prior to drug addition. As indicated in the figure, some cultures were incubated in the presence of the intracellular calcium chelator BAPTA/AM (20 μM for 20 min) prior to addition of bradykinin (BK) or C 2 -ceramide. For each treatment, 20–25 cells in three independent culture preparations were examined; each data point represents the mean ± SE.

    Article Snippet: Ceramide content was quantified within each experiment by using a standard curve established by assaying samples of authentic brain ceramide (50–400 pmol) processed and analyzed in parallel; the reference ceramide yielded an R f value of 0.25, as reported ( ).

    Techniques: Incubation, Fluorescence

    Effect of subcutaneous rIL-5 administration on eosinophil recruitment into challenged areas of skin. (a) Guinea-pigs were challenged with several doses of DNCB on the dorsal skin. Thereafter, 10 pmol/kg rIL-5 was injected s.c. into the dorsal surface of the left ear lobe 0 or 12 h after challenge. At 24 h after challenge, lesional skin was excised and the number of eosinophils was enumerated. Recombinant IL-5 administered 12 h after challenge induced a small but significant increase in the number of accumulating eosinophils in the dorsal skin challenged with 0·15% DNCB. (b) Guinea-pigs challenged on the right ear lobe with 0·15% DNCB were administered s.c. with several doses of rIL-5 into the left ear lobe 0 or 12 h after challenge. At 24 h after challenge, the right ear lobe was removed and the number of eosinophils was enumerated. Whereas rIL-5 (10 pmol/kg) injected 12 h after challenge remarkably enhanced eosinophil infiltration into the right ear lobe, tissue eosinophila was suppressed when rIL-5 was administered at the time of challenge (0 h). Each group consisted of at least four animals.

    Journal: Clinical and Experimental Immunology

    Article Title: Pathogenic roles of eosinophils in guinea-pig contact sensitivity: regulation of dermal eosinophilia with remotely administered IL-5

    doi: 10.1046/j.1365-2249.2000.01355.x

    Figure Lengend Snippet: Effect of subcutaneous rIL-5 administration on eosinophil recruitment into challenged areas of skin. (a) Guinea-pigs were challenged with several doses of DNCB on the dorsal skin. Thereafter, 10 pmol/kg rIL-5 was injected s.c. into the dorsal surface of the left ear lobe 0 or 12 h after challenge. At 24 h after challenge, lesional skin was excised and the number of eosinophils was enumerated. Recombinant IL-5 administered 12 h after challenge induced a small but significant increase in the number of accumulating eosinophils in the dorsal skin challenged with 0·15% DNCB. (b) Guinea-pigs challenged on the right ear lobe with 0·15% DNCB were administered s.c. with several doses of rIL-5 into the left ear lobe 0 or 12 h after challenge. At 24 h after challenge, the right ear lobe was removed and the number of eosinophils was enumerated. Whereas rIL-5 (10 pmol/kg) injected 12 h after challenge remarkably enhanced eosinophil infiltration into the right ear lobe, tissue eosinophila was suppressed when rIL-5 was administered at the time of challenge (0 h). Each group consisted of at least four animals.

    Article Snippet: Sensitized guinea-pigs were injected s.c. with several doses of human recombinant IL-5 (rIL-5) (0·1–10 pmol/kg) into the dorsal surface of the left ear lobe.

    Techniques: Injection, Recombinant

    Subcutaneous injection of rIL-5 induces transient peripheral blood eosinophilia.Recombinant IL-5 in 0·1 ml PBS was injected s.c. into the dorsal surface of the left ear lobe. Peripheral blood obtained from the retro-orbital plexus was serially sampled to determine the absolute eosinophil count. Each point is the mean of counts in four animals.

    Journal: Clinical and Experimental Immunology

    Article Title: Pathogenic roles of eosinophils in guinea-pig contact sensitivity: regulation of dermal eosinophilia with remotely administered IL-5

    doi: 10.1046/j.1365-2249.2000.01355.x

    Figure Lengend Snippet: Subcutaneous injection of rIL-5 induces transient peripheral blood eosinophilia.Recombinant IL-5 in 0·1 ml PBS was injected s.c. into the dorsal surface of the left ear lobe. Peripheral blood obtained from the retro-orbital plexus was serially sampled to determine the absolute eosinophil count. Each point is the mean of counts in four animals.

    Article Snippet: Sensitized guinea-pigs were injected s.c. with several doses of human recombinant IL-5 (rIL-5) (0·1–10 pmol/kg) into the dorsal surface of the left ear lobe.

    Techniques: Injection, Recombinant