The Experts below are selected from a list of 297 Experts worldwide ranked by ideXlab platform
Darlene A. Dartt - One of the best experts on this subject based on the ideXlab platform.
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Effect of overexpression of constitutively active PKCalpha on rat lacrimal gland Protein Secretion.
Investigative ophthalmology & visual science, 2004Co-Authors: Robin R. Hodges, Isabelle Raddassi, Driss Zoukhri, Alex Toker, Andrius Kazlauskas, Darlene A. DarttAbstract:PURPOSE. The lacrimal gland secretes water, electrolytes, and Protein into the tear film. Decreased Secretion from the lacrimal gland can lead to dry eye syndromes with deleterious effects on vision. Protein kinase C (PKC)- plays a major role in cholinergic- and 1-adrenergic‐induced Protein Secretion from the lacrimal gland. This study was undertaken to determine whether activation of PKC alone would induce lacrimal gland Protein Secretion by examining the effects of overexpression of constitutively active PKC. METHODS. Rat lacrimal gland acini were transduced with an adenovirus containing a gene for constitutively active PKC. Protein Secretion was measured in response to cholinergic and 1-adrenergic agonist stimulation. RESULTS. More than 84% of acinar cells were transduced, and PKC expression was increased 176-fold. Western blot analysis using an antibody to phosphorylated (activated) PKC indicated that the overexpressed PKC was active, and basal Secretion was increased. Cholinergic agonist‐stimulated Protein Secretion was not stimulated above basal Secretion, whereas 1-adrenergic-agonist‐stimulated Protein Secretion was increased in transduced acini. CONCLUSIONS. Basal lacrimal gland Protein Secretion can be stimulated by bypassing the release of neurotransmitters and activating PKC, possibly leading to the development of new treatments for dry eye syndromes. (Invest Ophthalmol Vis Sci. 2004;45:3974‐3981) DOI:10.1167/iovs.04-0508
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Lacrimal gland PKC isoforms are differentially involved in agonist-induced Protein Secretion
American Journal of Physiology-Cell Physiology, 1997Co-Authors: Driss Zoukhri, Robin R. Hodges, Alex Toker, Christian Sergheraert, Darlene A. DarttAbstract:In the present study, we have synthesized and N-myristoylated peptides derived from the pseudosubstrate sequences of Protein kinase C (PKC)-alpha, -delta, and -epsilon [Myr-PKC-alpha-(15-28), Myr-PKC-delta-(142-153), and Myr-PKC-epsilon-(149-164)], three isoforms present in rat lacrimal gland, and a peptide derived from the sequence of the endogenous inhibitor of Protein kinase A [Myr-PKI-(17-25)]. Lacrimal gland acini were preincubated for 60 min with the myristoylated peptides (10(-10) to 3 x 10(-7) M), then Protein Secretion was stimulated with a phorbol ester, phorbol 12,13-dibutyrate (10(-6) M); vasoactive intestinal peptide (10(-8) M); a cholinergic agonist, carbachol (10(-5) M); or an alpha 1-adrenergic agonist, phenylephrine (10(-4) M), for 20 min. In intact lacrimal gland acini, Myr-PKC-alpha-(15-28) inhibited phorbol 12,13-dibutyrate-induced Protein Secretion. This effect was not reproduced by the acetylated peptide or by the myristoylated PKI, which inhibited vasoactive intestinal peptide-induced Protein Secretion, a response mediated by Protein kinase A. Carbachol-induced Protein Secretion was inhibited by all three peptides. In contrast, phenylephrine-induced Protein Secretion was inhibited only by Myr-PKC-epsilon-(149-164), whereas Myr-PKC-alpha-(15-28) and Myr-PKC-delta-(142-153) had a stimulatory effect. None of these myristoylated peptides affected the calcium increase evoked by cholinergic or alpha 1-adrenergic agonists. We concluded that phorbol ester- and receptor-induced Protein Secretion involve different PKC isoforms in lacrimal gland.
Robin R. Hodges - One of the best experts on this subject based on the ideXlab platform.
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Effect of overexpression of constitutively active PKCalpha on rat lacrimal gland Protein Secretion.
Investigative ophthalmology & visual science, 2004Co-Authors: Robin R. Hodges, Isabelle Raddassi, Driss Zoukhri, Alex Toker, Andrius Kazlauskas, Darlene A. DarttAbstract:PURPOSE. The lacrimal gland secretes water, electrolytes, and Protein into the tear film. Decreased Secretion from the lacrimal gland can lead to dry eye syndromes with deleterious effects on vision. Protein kinase C (PKC)- plays a major role in cholinergic- and 1-adrenergic‐induced Protein Secretion from the lacrimal gland. This study was undertaken to determine whether activation of PKC alone would induce lacrimal gland Protein Secretion by examining the effects of overexpression of constitutively active PKC. METHODS. Rat lacrimal gland acini were transduced with an adenovirus containing a gene for constitutively active PKC. Protein Secretion was measured in response to cholinergic and 1-adrenergic agonist stimulation. RESULTS. More than 84% of acinar cells were transduced, and PKC expression was increased 176-fold. Western blot analysis using an antibody to phosphorylated (activated) PKC indicated that the overexpressed PKC was active, and basal Secretion was increased. Cholinergic agonist‐stimulated Protein Secretion was not stimulated above basal Secretion, whereas 1-adrenergic-agonist‐stimulated Protein Secretion was increased in transduced acini. CONCLUSIONS. Basal lacrimal gland Protein Secretion can be stimulated by bypassing the release of neurotransmitters and activating PKC, possibly leading to the development of new treatments for dry eye syndromes. (Invest Ophthalmol Vis Sci. 2004;45:3974‐3981) DOI:10.1167/iovs.04-0508
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Lacrimal gland PKC isoforms are differentially involved in agonist-induced Protein Secretion
American Journal of Physiology-Cell Physiology, 1997Co-Authors: Driss Zoukhri, Robin R. Hodges, Alex Toker, Christian Sergheraert, Darlene A. DarttAbstract:In the present study, we have synthesized and N-myristoylated peptides derived from the pseudosubstrate sequences of Protein kinase C (PKC)-alpha, -delta, and -epsilon [Myr-PKC-alpha-(15-28), Myr-PKC-delta-(142-153), and Myr-PKC-epsilon-(149-164)], three isoforms present in rat lacrimal gland, and a peptide derived from the sequence of the endogenous inhibitor of Protein kinase A [Myr-PKI-(17-25)]. Lacrimal gland acini were preincubated for 60 min with the myristoylated peptides (10(-10) to 3 x 10(-7) M), then Protein Secretion was stimulated with a phorbol ester, phorbol 12,13-dibutyrate (10(-6) M); vasoactive intestinal peptide (10(-8) M); a cholinergic agonist, carbachol (10(-5) M); or an alpha 1-adrenergic agonist, phenylephrine (10(-4) M), for 20 min. In intact lacrimal gland acini, Myr-PKC-alpha-(15-28) inhibited phorbol 12,13-dibutyrate-induced Protein Secretion. This effect was not reproduced by the acetylated peptide or by the myristoylated PKI, which inhibited vasoactive intestinal peptide-induced Protein Secretion, a response mediated by Protein kinase A. Carbachol-induced Protein Secretion was inhibited by all three peptides. In contrast, phenylephrine-induced Protein Secretion was inhibited only by Myr-PKC-epsilon-(149-164), whereas Myr-PKC-alpha-(15-28) and Myr-PKC-delta-(142-153) had a stimulatory effect. None of these myristoylated peptides affected the calcium increase evoked by cholinergic or alpha 1-adrenergic agonists. We concluded that phorbol ester- and receptor-induced Protein Secretion involve different PKC isoforms in lacrimal gland.
Driss Zoukhri - One of the best experts on this subject based on the ideXlab platform.
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Effect of overexpression of constitutively active PKCalpha on rat lacrimal gland Protein Secretion.
Investigative ophthalmology & visual science, 2004Co-Authors: Robin R. Hodges, Isabelle Raddassi, Driss Zoukhri, Alex Toker, Andrius Kazlauskas, Darlene A. DarttAbstract:PURPOSE. The lacrimal gland secretes water, electrolytes, and Protein into the tear film. Decreased Secretion from the lacrimal gland can lead to dry eye syndromes with deleterious effects on vision. Protein kinase C (PKC)- plays a major role in cholinergic- and 1-adrenergic‐induced Protein Secretion from the lacrimal gland. This study was undertaken to determine whether activation of PKC alone would induce lacrimal gland Protein Secretion by examining the effects of overexpression of constitutively active PKC. METHODS. Rat lacrimal gland acini were transduced with an adenovirus containing a gene for constitutively active PKC. Protein Secretion was measured in response to cholinergic and 1-adrenergic agonist stimulation. RESULTS. More than 84% of acinar cells were transduced, and PKC expression was increased 176-fold. Western blot analysis using an antibody to phosphorylated (activated) PKC indicated that the overexpressed PKC was active, and basal Secretion was increased. Cholinergic agonist‐stimulated Protein Secretion was not stimulated above basal Secretion, whereas 1-adrenergic-agonist‐stimulated Protein Secretion was increased in transduced acini. CONCLUSIONS. Basal lacrimal gland Protein Secretion can be stimulated by bypassing the release of neurotransmitters and activating PKC, possibly leading to the development of new treatments for dry eye syndromes. (Invest Ophthalmol Vis Sci. 2004;45:3974‐3981) DOI:10.1167/iovs.04-0508
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Lacrimal gland PKC isoforms are differentially involved in agonist-induced Protein Secretion
American Journal of Physiology-Cell Physiology, 1997Co-Authors: Driss Zoukhri, Robin R. Hodges, Alex Toker, Christian Sergheraert, Darlene A. DarttAbstract:In the present study, we have synthesized and N-myristoylated peptides derived from the pseudosubstrate sequences of Protein kinase C (PKC)-alpha, -delta, and -epsilon [Myr-PKC-alpha-(15-28), Myr-PKC-delta-(142-153), and Myr-PKC-epsilon-(149-164)], three isoforms present in rat lacrimal gland, and a peptide derived from the sequence of the endogenous inhibitor of Protein kinase A [Myr-PKI-(17-25)]. Lacrimal gland acini were preincubated for 60 min with the myristoylated peptides (10(-10) to 3 x 10(-7) M), then Protein Secretion was stimulated with a phorbol ester, phorbol 12,13-dibutyrate (10(-6) M); vasoactive intestinal peptide (10(-8) M); a cholinergic agonist, carbachol (10(-5) M); or an alpha 1-adrenergic agonist, phenylephrine (10(-4) M), for 20 min. In intact lacrimal gland acini, Myr-PKC-alpha-(15-28) inhibited phorbol 12,13-dibutyrate-induced Protein Secretion. This effect was not reproduced by the acetylated peptide or by the myristoylated PKI, which inhibited vasoactive intestinal peptide-induced Protein Secretion, a response mediated by Protein kinase A. Carbachol-induced Protein Secretion was inhibited by all three peptides. In contrast, phenylephrine-induced Protein Secretion was inhibited only by Myr-PKC-epsilon-(149-164), whereas Myr-PKC-alpha-(15-28) and Myr-PKC-delta-(142-153) had a stimulatory effect. None of these myristoylated peptides affected the calcium increase evoked by cholinergic or alpha 1-adrenergic agonists. We concluded that phorbol ester- and receptor-induced Protein Secretion involve different PKC isoforms in lacrimal gland.
Howard C. Hang - One of the best experts on this subject based on the ideXlab platform.
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Visible Fluorescence Detection of Type III Protein Secretion from Bacterial Pathogens
Journal of the American Chemical Society, 2010Co-Authors: Jacob S. Yount, Lun K. Tsou, Amy L. Kullas, Adrianus W. M. Van Der Velden, Pietro Domingues Dossa, Howard C. HangAbstract:Type III Protein Secretion is essential for many Gram-negative bacterial infections of host cells and an attractive target for new antibacterial drugs. Here, we describe a bacterial Protein effector-carboxypeptidase G2 (CPG2) reporter system for fluorescence and visible detection of type III Protein Secretion in Salmonella typhimurium. This system provides a general method for measuring Protein expression and Secretion as well as a high-throughput and quantitative assay for analyzing type III Protein Secretion inhibitors.
Alex Toker - One of the best experts on this subject based on the ideXlab platform.
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Effect of overexpression of constitutively active PKCalpha on rat lacrimal gland Protein Secretion.
Investigative ophthalmology & visual science, 2004Co-Authors: Robin R. Hodges, Isabelle Raddassi, Driss Zoukhri, Alex Toker, Andrius Kazlauskas, Darlene A. DarttAbstract:PURPOSE. The lacrimal gland secretes water, electrolytes, and Protein into the tear film. Decreased Secretion from the lacrimal gland can lead to dry eye syndromes with deleterious effects on vision. Protein kinase C (PKC)- plays a major role in cholinergic- and 1-adrenergic‐induced Protein Secretion from the lacrimal gland. This study was undertaken to determine whether activation of PKC alone would induce lacrimal gland Protein Secretion by examining the effects of overexpression of constitutively active PKC. METHODS. Rat lacrimal gland acini were transduced with an adenovirus containing a gene for constitutively active PKC. Protein Secretion was measured in response to cholinergic and 1-adrenergic agonist stimulation. RESULTS. More than 84% of acinar cells were transduced, and PKC expression was increased 176-fold. Western blot analysis using an antibody to phosphorylated (activated) PKC indicated that the overexpressed PKC was active, and basal Secretion was increased. Cholinergic agonist‐stimulated Protein Secretion was not stimulated above basal Secretion, whereas 1-adrenergic-agonist‐stimulated Protein Secretion was increased in transduced acini. CONCLUSIONS. Basal lacrimal gland Protein Secretion can be stimulated by bypassing the release of neurotransmitters and activating PKC, possibly leading to the development of new treatments for dry eye syndromes. (Invest Ophthalmol Vis Sci. 2004;45:3974‐3981) DOI:10.1167/iovs.04-0508
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Lacrimal gland PKC isoforms are differentially involved in agonist-induced Protein Secretion
American Journal of Physiology-Cell Physiology, 1997Co-Authors: Driss Zoukhri, Robin R. Hodges, Alex Toker, Christian Sergheraert, Darlene A. DarttAbstract:In the present study, we have synthesized and N-myristoylated peptides derived from the pseudosubstrate sequences of Protein kinase C (PKC)-alpha, -delta, and -epsilon [Myr-PKC-alpha-(15-28), Myr-PKC-delta-(142-153), and Myr-PKC-epsilon-(149-164)], three isoforms present in rat lacrimal gland, and a peptide derived from the sequence of the endogenous inhibitor of Protein kinase A [Myr-PKI-(17-25)]. Lacrimal gland acini were preincubated for 60 min with the myristoylated peptides (10(-10) to 3 x 10(-7) M), then Protein Secretion was stimulated with a phorbol ester, phorbol 12,13-dibutyrate (10(-6) M); vasoactive intestinal peptide (10(-8) M); a cholinergic agonist, carbachol (10(-5) M); or an alpha 1-adrenergic agonist, phenylephrine (10(-4) M), for 20 min. In intact lacrimal gland acini, Myr-PKC-alpha-(15-28) inhibited phorbol 12,13-dibutyrate-induced Protein Secretion. This effect was not reproduced by the acetylated peptide or by the myristoylated PKI, which inhibited vasoactive intestinal peptide-induced Protein Secretion, a response mediated by Protein kinase A. Carbachol-induced Protein Secretion was inhibited by all three peptides. In contrast, phenylephrine-induced Protein Secretion was inhibited only by Myr-PKC-epsilon-(149-164), whereas Myr-PKC-alpha-(15-28) and Myr-PKC-delta-(142-153) had a stimulatory effect. None of these myristoylated peptides affected the calcium increase evoked by cholinergic or alpha 1-adrenergic agonists. We concluded that phorbol ester- and receptor-induced Protein Secretion involve different PKC isoforms in lacrimal gland.
