G. Kunos - Academia.edu (original) (raw)
Papers by G. Kunos
Journal of Biological Chemistry, 1988
Coculturing IM9 human lymphocytes and A549 human lung adenocarcinoma cells results in a 2-3-fold ... more Coculturing IM9 human lymphocytes and A549 human lung adenocarcinoma cells results in a 2-3-fold increase in the density of beta-adrenergic receptors in the latter, as quantified by 125I-cyanopindolol binding. Lymphocyte-conditioned medium (LCM) has the same effect, which is moderately sensitive to heat, is retained by ultrafiltration over a Mr 10,000 cut-off filter, and is reduced by trypsin treatment or by preincubation of lymphocytes with 0.3 micrograms/ml cycloheximide. Treatment of lung cells with cycloheximide also prevents the effect of LCM. Glucocorticoids, which also increase beta-receptor density in A549 cells, markedly potentiate the effect of LCM. Gel permeation high pressure liquid chromatography of LCM yields three peaks of biological activity with Mr 70,000, 35,000, and 15,000. Monocytic interleukin-1 (IL-1) mimics the effect of LCM in that it increases beta-receptor density in A549 cells (EC50 0.3 pM), and its effect is potentiated by cortisol. Recombinant IL-1 alpha is somewhat more potent than IL-1 beta, while interleukin-2 and interferon-alpha are ineffective. Tumor necrosis factor alpha causes a small increase in beta-receptors, which is not influenced by glucocorticoids. A polyclonal anti-IL-1 antibody inhibits the effect of IL-1 and the effect of the 15-kDa but not the 35- and 70-kDa fractions of LCM. The activity of the latter two fractions is also unaffected by anti-tumor necrosis factor alpha antibody. These results indicate that lymphocytes release protein factors including IL-1 that up-regulate pulmonary beta-adrenergic receptors by an action that involves protein synthesis. The possible relevance of this regulatory mechanism for the pathomechanism of certain respiratory diseases is discussed.
A68. PARENCHYMA/INTERSTITIAL LUNG DISEASE, 2020
RATIONALE: Cannabinoid 1 receptor (CB 1 R) has emerged as a contributor to pulmonary fibrosis (PF... more RATIONALE: Cannabinoid 1 receptor (CB 1 R) has emerged as a contributor to pulmonary fibrosis (PF). Additionally, a significant increase in the anandamide (AEA) content of BALF occurs in both IPF patients and in mice with experimental PF and is negatively correlated with pulmonary function in IPF patients. Lung and plasma levels of chemokine (C-X-C motif) ligand 13 (CXCL13), known as B lymphocyte chemoattractant, have been recently identified as a prognostic biomarker for IPF. Here, we explored the role of lung and myeloid cell CB 1 Rs in regulating CXCL13 production and lymphocyte infiltration in bleomycin (BLEO)-induced PF in
A29. EMERGING CONCEPTS IN LUNG FIBROSIS, 2019
Biyofilmler. gerek tıbbi cihaz ve biyomateryaller üzerinde gerekse konakçı epitel hücreleri ve mu... more Biyofilmler. gerek tıbbi cihaz ve biyomateryaller üzerinde gerekse konakçı epitel hücreleri ve mukozal yüzeylerde oluşabilen ve pek çok farklı hastalıkta rol oynayan mikroekosistemlerdir. Biyofilm oluşumunu belirleme yöntemleri, biyofilm enfeksiyonlarının engellenebilmesi için oldukça önemlidir. Biyofilm oluşumunu saptamada sıklıkla kullanılan kolorimetrik esaslı yöntemlerin çeşitli dezavantajları bulunduğundan, standart, hızlı ve güvenilir yöntemlere gereksinim duyulmaktadır. Biyofilm saptama amacıyla da kullanılabilen çeşitli yeni teknolojiler geliştirilmiştir. Bunlar arasında, elektrik akımına dirençteki değişimin saptandığı "gerçek zamanlı hücre analizi" yöntemi, kızılötesi ışınların penetrasyonundaki değişimin ölçüldüğü "fiber-optik sensör" yöntemi ve biyofilm matriks komponentlerini saptamaya dayalı "luminisan konjue oligotiyofen" yöntemi umut vadetmektedir. Bu yöntemler ile daha duyarlı, hızlı ve kantitatif olarak biyofilm yapısı belirlenebilmektedir. Planktonik hâldeki bakterilerin antibiyotik direncinin yanı sıra, biyofilm yapısından kaynaklanan direnç, biyofilm enfeksiyonlarında tedaviyi daha da zorlaştırmaktadır. Biyofilm direncinin multifaktöriyel bir olay olduğu ve birden fazla mekanizmanın eşzamanlı etkisiyle ortaya çıktığı düşünülmektedir. Derlememizde, bakteriyel biyofilmlerin oluşumu ile bunu etkileyen faktörler, biyofilm saptama yöntemleri, biyofilm enfeksiyonları ve biyofilm direncinde rol oynayan etkenler incelenmektedir.
Molecular Pharmacology, 1997
cAMP markedly increases ␣ 1B adrenergic receptor (␣ 1B-AR) expression in FRTL-5 and PC C13 rat th... more cAMP markedly increases ␣ 1B adrenergic receptor (␣ 1B-AR) expression in FRTL-5 and PC C13 rat thyroid cells, DDT 1 MF-2 smooth muscle cells, primary rat hepatocytes, and K9 rat liver cells. Here, we used DDT 1 MF-2 cells to evaluate further the mechanisms by which cAMP stimulates ␣ 1B-AR expression. Receptor binding assays, Northern blotting, and nuclear run-on analyses demonstrated that forskolin (1 M) in the presence of isobutylmethylxanthine (0.25 mM) increased ␣ 1B-AR numbers, mRNA level, and gene transcription rate by 2.3 Ϯ 0.2-, 2.5 Ϯ 0.3-, and 3.5 Ϯ 0.2-fold over control, respectively. Dibutyryl cAMP (1 mM) plus isobutylmethylxanthine (0.25 mM) also enhanced ␣ 1B-AR density by 2.7 Ϯ 0.1-fold over control. Further experiments demonstrated that the induction of ␣ 1B-AR by forskolin requires new protein synthesis and is protein kinase A dependent. In DDT 1 MF-2 cells transfected with ␣ 1B-AR gene P2 promoter/CAT constructs, both forskolin and dibutyryl
Nature, 1997
Anandamide, an endogenous cannabinoid ligand 1 , binds to CB 1 cannabinoid receptors 2 in the bra... more Anandamide, an endogenous cannabinoid ligand 1 , binds to CB 1 cannabinoid receptors 2 in the brain and mimics the neurobehavioural actions of marijuana 3,4. Cannabinoids and anandamide also elicit hypotension mediated by peripheral CB 1 receptors 5-8. Here we report that a selective CB 1 receptor antagonist, SR141716A 9 , elicits an increase in blood pressure in rats subjected to haemorrhagic shock, whereas similar treatment of normotensive rats or intracerebroventricular administration of the antagonist during shock do not affect blood pressure. Blood from haemorrhaged rats causes hypotension in normal rats, which can be prevented by SR141716A but not by inhibition of nitric oxide synthase in the recipient. Macrophages and platelets from haemorrhaged rats elicit CB 1 receptor-mediated hypotension in normotensive recipients, and incorporate arachidonic acid or ethanolamine into a product that co-elutes with anandamide on reverse-phase high-performance liquid chromatography. Also, macrophages from control rats stimulated with ionomycin or bacterial phospholipase D produce anandamide, as identified by gas chromatography and mass spectrometry. These findings indicate that activation of peripheral CB 1 cannabinoid receptors contributes to haemorrhagic hypotension, and anandamide produced by macrophages may be a mediator of this effect. Rats anaesthetized with urethane were subjected to standardized, stepwise bleeding until mean blood pressure stabilized at 40 mm Hg (refs 10, 11). Blood pressure remained stable at this level for 25-35 min, after which it started to decline and the animal died. At the beginning of the stable hypotensive phase, some rats received 3 mg kg −1 of SR141716A intravenously (i.v.), which caused a marked and prolonged pressor response that lasted 20-30 min (Fig. 1). The
Proceedings of the National Academy of Sciences, 1999
Cannabinoids, including the endogenous ligand arachidonyl ethanolamide (anandamide), elicit not o... more Cannabinoids, including the endogenous ligand arachidonyl ethanolamide (anandamide), elicit not only neurobehavioral but also cardiovascular effects. Two cannabinoid receptors, CB1 and CB2, have been cloned, and studies with the selective CB1 receptor antagonist SR141716A have implicated peripherally located CB1 receptors in the hypotensive action of cannabinoids. In rat mesenteric arteries, anandamide-induced vasodilation is inhibited by SR141716A, but other potent CB1 receptor agonists, such as HU-210, do not cause vasodilation, which implicates an as-yet-unidentified receptor in this effect. Here we show that “abnormal cannabidiol” (Abn-cbd) is a neurobehaviorally inactive cannabinoid that does not bind to CB1 receptors, yet causes SR141716A-sensitive hypotension and mesenteric vasodilation in wild-type mice and in mice lacking CB1 receptors or both CB1 and CB2 receptors. Hypotension by Abn-cbd is also inhibited by cannabidiol (20 μg/g), which does not influence anandamide- or HU...
Life Sciences, 1979
ABSTRACT
Free Radical Biology and Medicine, 2014
Orvosi hetilap, Jan 24, 1983
Orvosi hetilap, Jan 23, 1983
The Journal of pharmacology and experimental therapeutics, 1982
The effects of prolonged in vivo infusion of isoproterenol (400 micrograms/kg/hr) or norepinephri... more The effects of prolonged in vivo infusion of isoproterenol (400 micrograms/kg/hr) or norepinephrine (200 micrograms/kg/hr) from a minipump on the physiological reactivity and binding properties of cardiac beta and alpha-1 adrenoceptors were tested in rats. Infusion of either catecholamine significantly reduced the in vitro inotropic and chronotropic potency of isoproterenol in isolated left and right atria, respectively; desensitization was near maximal as early as after 2 hr of infusion. No significant change in the density of [3H]dihydroalprenolol-labeled beta receptors was evident at this time point in either atrial or ventricular tissue, although isoproterenol did decrease binding site density after 7 days of infusion. There was no change in the binding affinity or physiological blocking potency of dihydroalprenolol after isoproterenol infusion. The inotropic potency of phenylephrine in the presence of dihydroalprenolol was unaffected by infusion of either isoproterenol or norep...
Arzneimittel-Forschung, 1968
Adrenergic Receptor Protocols
ABSTRACT Adrenergic receptors (ARs) are G-protein-coupled transmembrane glyco-proteins that play ... more ABSTRACT Adrenergic receptors (ARs) are G-protein-coupled transmembrane glyco-proteins that play a key role in mediating the sympathoadrenal response to stress. Pharmacological studies have suggested the existence of multiple adrenergic receptor subtypes and sub-subtypes, and to date, nine distinct AR genes have been identified by molecular cloning, including three α1-AR subtypes (α1A/D, α1B, and α1C), three α2-AR (α2A, α2B, and α2C) and three β-AR (β1, β2, and β3) (1,2). The expression of these receptors is regulated in a complex tissue-specific manner. Such regulation by hormones or developmental factors has been shown to occur at the transcriptional level under many of these conditions. To understand the molecular mechanisms involved in regulating the transcription of the AR under a variety of physiological and pathological conditions, many AR genes have been isolated. After the promoter sequences of receptor genes have been defined through the use of chloroamphenicol acetyltransferase (CAT) reporter assays, a computer-based search of the Sitedata database of previously identified cis-elements can be used to check whether these promoter sequences contain known sequence-specific protein binding sites. To determine whether, in a given tissue, a protein does in fact bind to a consensus recognition sequence on the DNA and/or to identify novel cis-elements, DNase I footprinting techniques can be used. Once protein binding to DNA is verified by using this technique, a number of other methods, including DNA-gel mobility shift and supershift assays and mutational analyses can be used to characterize the trans-acting factors bound to any specific site.
Journal of Biological Chemistry, 1988
Coculturing IM9 human lymphocytes and A549 human lung adenocarcinoma cells results in a 2-3-fold ... more Coculturing IM9 human lymphocytes and A549 human lung adenocarcinoma cells results in a 2-3-fold increase in the density of beta-adrenergic receptors in the latter, as quantified by 125I-cyanopindolol binding. Lymphocyte-conditioned medium (LCM) has the same effect, which is moderately sensitive to heat, is retained by ultrafiltration over a Mr 10,000 cut-off filter, and is reduced by trypsin treatment or by preincubation of lymphocytes with 0.3 micrograms/ml cycloheximide. Treatment of lung cells with cycloheximide also prevents the effect of LCM. Glucocorticoids, which also increase beta-receptor density in A549 cells, markedly potentiate the effect of LCM. Gel permeation high pressure liquid chromatography of LCM yields three peaks of biological activity with Mr 70,000, 35,000, and 15,000. Monocytic interleukin-1 (IL-1) mimics the effect of LCM in that it increases beta-receptor density in A549 cells (EC50 0.3 pM), and its effect is potentiated by cortisol. Recombinant IL-1 alpha is somewhat more potent than IL-1 beta, while interleukin-2 and interferon-alpha are ineffective. Tumor necrosis factor alpha causes a small increase in beta-receptors, which is not influenced by glucocorticoids. A polyclonal anti-IL-1 antibody inhibits the effect of IL-1 and the effect of the 15-kDa but not the 35- and 70-kDa fractions of LCM. The activity of the latter two fractions is also unaffected by anti-tumor necrosis factor alpha antibody. These results indicate that lymphocytes release protein factors including IL-1 that up-regulate pulmonary beta-adrenergic receptors by an action that involves protein synthesis. The possible relevance of this regulatory mechanism for the pathomechanism of certain respiratory diseases is discussed.
A68. PARENCHYMA/INTERSTITIAL LUNG DISEASE, 2020
RATIONALE: Cannabinoid 1 receptor (CB 1 R) has emerged as a contributor to pulmonary fibrosis (PF... more RATIONALE: Cannabinoid 1 receptor (CB 1 R) has emerged as a contributor to pulmonary fibrosis (PF). Additionally, a significant increase in the anandamide (AEA) content of BALF occurs in both IPF patients and in mice with experimental PF and is negatively correlated with pulmonary function in IPF patients. Lung and plasma levels of chemokine (C-X-C motif) ligand 13 (CXCL13), known as B lymphocyte chemoattractant, have been recently identified as a prognostic biomarker for IPF. Here, we explored the role of lung and myeloid cell CB 1 Rs in regulating CXCL13 production and lymphocyte infiltration in bleomycin (BLEO)-induced PF in
A29. EMERGING CONCEPTS IN LUNG FIBROSIS, 2019
Biyofilmler. gerek tıbbi cihaz ve biyomateryaller üzerinde gerekse konakçı epitel hücreleri ve mu... more Biyofilmler. gerek tıbbi cihaz ve biyomateryaller üzerinde gerekse konakçı epitel hücreleri ve mukozal yüzeylerde oluşabilen ve pek çok farklı hastalıkta rol oynayan mikroekosistemlerdir. Biyofilm oluşumunu belirleme yöntemleri, biyofilm enfeksiyonlarının engellenebilmesi için oldukça önemlidir. Biyofilm oluşumunu saptamada sıklıkla kullanılan kolorimetrik esaslı yöntemlerin çeşitli dezavantajları bulunduğundan, standart, hızlı ve güvenilir yöntemlere gereksinim duyulmaktadır. Biyofilm saptama amacıyla da kullanılabilen çeşitli yeni teknolojiler geliştirilmiştir. Bunlar arasında, elektrik akımına dirençteki değişimin saptandığı "gerçek zamanlı hücre analizi" yöntemi, kızılötesi ışınların penetrasyonundaki değişimin ölçüldüğü "fiber-optik sensör" yöntemi ve biyofilm matriks komponentlerini saptamaya dayalı "luminisan konjue oligotiyofen" yöntemi umut vadetmektedir. Bu yöntemler ile daha duyarlı, hızlı ve kantitatif olarak biyofilm yapısı belirlenebilmektedir. Planktonik hâldeki bakterilerin antibiyotik direncinin yanı sıra, biyofilm yapısından kaynaklanan direnç, biyofilm enfeksiyonlarında tedaviyi daha da zorlaştırmaktadır. Biyofilm direncinin multifaktöriyel bir olay olduğu ve birden fazla mekanizmanın eşzamanlı etkisiyle ortaya çıktığı düşünülmektedir. Derlememizde, bakteriyel biyofilmlerin oluşumu ile bunu etkileyen faktörler, biyofilm saptama yöntemleri, biyofilm enfeksiyonları ve biyofilm direncinde rol oynayan etkenler incelenmektedir.
Molecular Pharmacology, 1997
cAMP markedly increases ␣ 1B adrenergic receptor (␣ 1B-AR) expression in FRTL-5 and PC C13 rat th... more cAMP markedly increases ␣ 1B adrenergic receptor (␣ 1B-AR) expression in FRTL-5 and PC C13 rat thyroid cells, DDT 1 MF-2 smooth muscle cells, primary rat hepatocytes, and K9 rat liver cells. Here, we used DDT 1 MF-2 cells to evaluate further the mechanisms by which cAMP stimulates ␣ 1B-AR expression. Receptor binding assays, Northern blotting, and nuclear run-on analyses demonstrated that forskolin (1 M) in the presence of isobutylmethylxanthine (0.25 mM) increased ␣ 1B-AR numbers, mRNA level, and gene transcription rate by 2.3 Ϯ 0.2-, 2.5 Ϯ 0.3-, and 3.5 Ϯ 0.2-fold over control, respectively. Dibutyryl cAMP (1 mM) plus isobutylmethylxanthine (0.25 mM) also enhanced ␣ 1B-AR density by 2.7 Ϯ 0.1-fold over control. Further experiments demonstrated that the induction of ␣ 1B-AR by forskolin requires new protein synthesis and is protein kinase A dependent. In DDT 1 MF-2 cells transfected with ␣ 1B-AR gene P2 promoter/CAT constructs, both forskolin and dibutyryl
Nature, 1997
Anandamide, an endogenous cannabinoid ligand 1 , binds to CB 1 cannabinoid receptors 2 in the bra... more Anandamide, an endogenous cannabinoid ligand 1 , binds to CB 1 cannabinoid receptors 2 in the brain and mimics the neurobehavioural actions of marijuana 3,4. Cannabinoids and anandamide also elicit hypotension mediated by peripheral CB 1 receptors 5-8. Here we report that a selective CB 1 receptor antagonist, SR141716A 9 , elicits an increase in blood pressure in rats subjected to haemorrhagic shock, whereas similar treatment of normotensive rats or intracerebroventricular administration of the antagonist during shock do not affect blood pressure. Blood from haemorrhaged rats causes hypotension in normal rats, which can be prevented by SR141716A but not by inhibition of nitric oxide synthase in the recipient. Macrophages and platelets from haemorrhaged rats elicit CB 1 receptor-mediated hypotension in normotensive recipients, and incorporate arachidonic acid or ethanolamine into a product that co-elutes with anandamide on reverse-phase high-performance liquid chromatography. Also, macrophages from control rats stimulated with ionomycin or bacterial phospholipase D produce anandamide, as identified by gas chromatography and mass spectrometry. These findings indicate that activation of peripheral CB 1 cannabinoid receptors contributes to haemorrhagic hypotension, and anandamide produced by macrophages may be a mediator of this effect. Rats anaesthetized with urethane were subjected to standardized, stepwise bleeding until mean blood pressure stabilized at 40 mm Hg (refs 10, 11). Blood pressure remained stable at this level for 25-35 min, after which it started to decline and the animal died. At the beginning of the stable hypotensive phase, some rats received 3 mg kg −1 of SR141716A intravenously (i.v.), which caused a marked and prolonged pressor response that lasted 20-30 min (Fig. 1). The
Proceedings of the National Academy of Sciences, 1999
Cannabinoids, including the endogenous ligand arachidonyl ethanolamide (anandamide), elicit not o... more Cannabinoids, including the endogenous ligand arachidonyl ethanolamide (anandamide), elicit not only neurobehavioral but also cardiovascular effects. Two cannabinoid receptors, CB1 and CB2, have been cloned, and studies with the selective CB1 receptor antagonist SR141716A have implicated peripherally located CB1 receptors in the hypotensive action of cannabinoids. In rat mesenteric arteries, anandamide-induced vasodilation is inhibited by SR141716A, but other potent CB1 receptor agonists, such as HU-210, do not cause vasodilation, which implicates an as-yet-unidentified receptor in this effect. Here we show that “abnormal cannabidiol” (Abn-cbd) is a neurobehaviorally inactive cannabinoid that does not bind to CB1 receptors, yet causes SR141716A-sensitive hypotension and mesenteric vasodilation in wild-type mice and in mice lacking CB1 receptors or both CB1 and CB2 receptors. Hypotension by Abn-cbd is also inhibited by cannabidiol (20 μg/g), which does not influence anandamide- or HU...
Life Sciences, 1979
ABSTRACT
Free Radical Biology and Medicine, 2014
Orvosi hetilap, Jan 24, 1983
Orvosi hetilap, Jan 23, 1983
The Journal of pharmacology and experimental therapeutics, 1982
The effects of prolonged in vivo infusion of isoproterenol (400 micrograms/kg/hr) or norepinephri... more The effects of prolonged in vivo infusion of isoproterenol (400 micrograms/kg/hr) or norepinephrine (200 micrograms/kg/hr) from a minipump on the physiological reactivity and binding properties of cardiac beta and alpha-1 adrenoceptors were tested in rats. Infusion of either catecholamine significantly reduced the in vitro inotropic and chronotropic potency of isoproterenol in isolated left and right atria, respectively; desensitization was near maximal as early as after 2 hr of infusion. No significant change in the density of [3H]dihydroalprenolol-labeled beta receptors was evident at this time point in either atrial or ventricular tissue, although isoproterenol did decrease binding site density after 7 days of infusion. There was no change in the binding affinity or physiological blocking potency of dihydroalprenolol after isoproterenol infusion. The inotropic potency of phenylephrine in the presence of dihydroalprenolol was unaffected by infusion of either isoproterenol or norep...
Arzneimittel-Forschung, 1968
Adrenergic Receptor Protocols
ABSTRACT Adrenergic receptors (ARs) are G-protein-coupled transmembrane glyco-proteins that play ... more ABSTRACT Adrenergic receptors (ARs) are G-protein-coupled transmembrane glyco-proteins that play a key role in mediating the sympathoadrenal response to stress. Pharmacological studies have suggested the existence of multiple adrenergic receptor subtypes and sub-subtypes, and to date, nine distinct AR genes have been identified by molecular cloning, including three α1-AR subtypes (α1A/D, α1B, and α1C), three α2-AR (α2A, α2B, and α2C) and three β-AR (β1, β2, and β3) (1,2). The expression of these receptors is regulated in a complex tissue-specific manner. Such regulation by hormones or developmental factors has been shown to occur at the transcriptional level under many of these conditions. To understand the molecular mechanisms involved in regulating the transcription of the AR under a variety of physiological and pathological conditions, many AR genes have been isolated. After the promoter sequences of receptor genes have been defined through the use of chloroamphenicol acetyltransferase (CAT) reporter assays, a computer-based search of the Sitedata database of previously identified cis-elements can be used to check whether these promoter sequences contain known sequence-specific protein binding sites. To determine whether, in a given tissue, a protein does in fact bind to a consensus recognition sequence on the DNA and/or to identify novel cis-elements, DNase I footprinting techniques can be used. Once protein binding to DNA is verified by using this technique, a number of other methods, including DNA-gel mobility shift and supershift assays and mutational analyses can be used to characterize the trans-acting factors bound to any specific site.