High turnover rate of central histaminergic system in patients with Down syndrome and Alzheimer disease

    Risultato della ricerca: Paper

    Abstract

    HIGH TURNOVER RATE OF CENTRAL HISTAMINERGIC SYSTEM IN PATIENTS WITH DOWN SYNDROME AND ALZHEIMER DISEASE Maria Concetta Gueli Dipartimento di Biomedicina Sperimentale e Neuroscienze Cliniche (BioNEC), Università degli Studi di Palermo It is well confirmed that a strong relationship exists between Down’s syndrome (DS) and Alzheimer’s disease (AD). Neurochemical investigations reported that many central neurotransmitter systems are similarly affected in aging Down and in Alzheimer patients, respectively. Airaksinem et al. (1) found numerous neurofibrillary tangles in the tuberomammillary area of the hypothalamus, where cell bodies of histaminergic neurons are located. While Mazurkiewicz-Kwilecki et al. (2) found deficits of the endogenous diamine, Cacabelos et al. (3) reported an increase of central histamine levels. In the present study, in order to test whether AD-like neuropathological changes involve the central histaminergic system, we measured the concentration of histamine, histidine as well as the activity of histidine decarboxylase (HDC) and histamine-Nmethyltransferase (HMT) in temporal cortex (TC) of aging Down, Alzheimer and control patients. Post-mortem samples (temporal cortex, TC; grey matter) of AD neuropathologically confirmed cases (72.1 ± 7.6 years old), of karyotyped patients with DS (56.1 ± 7.1 years old), and control adults (72,7 ± 9.7 years old) were obtained from the MRC London Brain Bank for Neurodegenerative Diseases, Department of Neuropathology, Institute of psychiatry, London, U.K. Each block of brain tissue from AD, DS and controls were thawed on ice and homogenized in ice-cold HDC-solution of 0.1 M sodium phosphate buffer (pH 6.8) containing dithiothreitol and antipain protease inhibitor. Homogenates of brain specimens were centrifuged at 12,000 x g for 20 min at 4°C. The supernatants were poured into CENTRIPEP-3 concentrators (Amicon), and centrifuged at 2,000 x g for two 10 min periods at 4°C. The clear extracts were stored in small quantities in Eppendorf tubes at -80°C until analysis. HDC activity has been measured with the procedure described by Gueli et al. (4) and briefly summarized. Extract aliquots were pre-incubated for 10 min with HDC assay-solution (0.1 M PBS, 0.2 mM DTT, 0.01 mM PLP, 0.1 mM Aminoguanidine), then incubation was started by adding 0.5 mM Lhistidine for 0-3 h at 37°C. At the established times, the reactions were stopped with 60 % ice-cold PCA, and stored overnight. Finally, the reaction mixture was centrifuged at 19,000 x g for 30 min at 4°C. The supernatants were withdrawn and filtered (0.45 mm Millipore filter). The HPLC system consisted of a 600E Waters pump with a Waters 474 scanning fluorescence detector (ex 350 nm, em 450 nm). Chromatograms and calculations were performed by Empower TM2 Data Software. Histamine was separated and quantified after pre-column derivatization with Shore’s o-phthalaldehyde reaction (5), using a Spherisorb ODS2 analytical column, particle size 3 mm (20 x 0.46 cm,) (Waters, Milano), a 10 L injection volume, and a mobile phase of methanol, 20 mmol/ L sodium acetate in water, acetic acid (55:43:2 v/v) and 0.33 mmol/L 1- octanesulfonic acid sodium salt. The flow rate was 1.0 ml/min. In order to measure HMT activity brain tissue was disperged with a glass Teflon homogenizer in 0.1 M PBS (pH 7.2). After centrifugation the supernatant was used for the radioenzymatic assay (6). Histidine contents were measured using the procedure described by Borum (7). We observed a increase of histamine levels in temporal cortex of AD (+15%) patients. Down brains also showed a mild increase of the endogenous diamine concentration (+8%). HDC activity in both groups of diseased brains was significantly increased compared
    Lingua originaleEnglish
    Stato di pubblicazionePublished - 2013

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    Histidine Decarboxylase
    Down Syndrome
    Temporal Lobe
    Alzheimer Disease
    Histamine
    Ice
    Diamines
    Histidine
    Water
    Brain
    Antipain
    Sodium Acetate
    Passive Cutaneous Anaphylaxis
    Neurofibrillary Tangles
    Dithiothreitol
    Polytetrafluoroethylene
    Brain Diseases
    Protease Inhibitors
    Centrifugation
    Particle Size

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    title = "High turnover rate of central histaminergic system in patients with Down syndrome and Alzheimer disease",
    abstract = "HIGH TURNOVER RATE OF CENTRAL HISTAMINERGIC SYSTEM IN PATIENTS WITH DOWN SYNDROME AND ALZHEIMER DISEASE Maria Concetta Gueli Dipartimento di Biomedicina Sperimentale e Neuroscienze Cliniche (BioNEC), Universit{\`a} degli Studi di Palermo It is well confirmed that a strong relationship exists between Down’s syndrome (DS) and Alzheimer’s disease (AD). Neurochemical investigations reported that many central neurotransmitter systems are similarly affected in aging Down and in Alzheimer patients, respectively. Airaksinem et al. (1) found numerous neurofibrillary tangles in the tuberomammillary area of the hypothalamus, where cell bodies of histaminergic neurons are located. While Mazurkiewicz-Kwilecki et al. (2) found deficits of the endogenous diamine, Cacabelos et al. (3) reported an increase of central histamine levels. In the present study, in order to test whether AD-like neuropathological changes involve the central histaminergic system, we measured the concentration of histamine, histidine as well as the activity of histidine decarboxylase (HDC) and histamine-Nmethyltransferase (HMT) in temporal cortex (TC) of aging Down, Alzheimer and control patients. Post-mortem samples (temporal cortex, TC; grey matter) of AD neuropathologically confirmed cases (72.1 ± 7.6 years old), of karyotyped patients with DS (56.1 ± 7.1 years old), and control adults (72,7 ± 9.7 years old) were obtained from the MRC London Brain Bank for Neurodegenerative Diseases, Department of Neuropathology, Institute of psychiatry, London, U.K. Each block of brain tissue from AD, DS and controls were thawed on ice and homogenized in ice-cold HDC-solution of 0.1 M sodium phosphate buffer (pH 6.8) containing dithiothreitol and antipain protease inhibitor. Homogenates of brain specimens were centrifuged at 12,000 x g for 20 min at 4°C. The supernatants were poured into CENTRIPEP-3 concentrators (Amicon), and centrifuged at 2,000 x g for two 10 min periods at 4°C. The clear extracts were stored in small quantities in Eppendorf tubes at -80°C until analysis. HDC activity has been measured with the procedure described by Gueli et al. (4) and briefly summarized. Extract aliquots were pre-incubated for 10 min with HDC assay-solution (0.1 M PBS, 0.2 mM DTT, 0.01 mM PLP, 0.1 mM Aminoguanidine), then incubation was started by adding 0.5 mM Lhistidine for 0-3 h at 37°C. At the established times, the reactions were stopped with 60 {\%} ice-cold PCA, and stored overnight. Finally, the reaction mixture was centrifuged at 19,000 x g for 30 min at 4°C. The supernatants were withdrawn and filtered (0.45 mm Millipore filter). The HPLC system consisted of a 600E Waters pump with a Waters 474 scanning fluorescence detector (ex 350 nm, em 450 nm). Chromatograms and calculations were performed by Empower TM2 Data Software. Histamine was separated and quantified after pre-column derivatization with Shore’s o-phthalaldehyde reaction (5), using a Spherisorb ODS2 analytical column, particle size 3 mm (20 x 0.46 cm,) (Waters, Milano), a 10 L injection volume, and a mobile phase of methanol, 20 mmol/ L sodium acetate in water, acetic acid (55:43:2 v/v) and 0.33 mmol/L 1- octanesulfonic acid sodium salt. The flow rate was 1.0 ml/min. In order to measure HMT activity brain tissue was disperged with a glass Teflon homogenizer in 0.1 M PBS (pH 7.2). After centrifugation the supernatant was used for the radioenzymatic assay (6). Histidine contents were measured using the procedure described by Borum (7). We observed a increase of histamine levels in temporal cortex of AD (+15{\%}) patients. Down brains also showed a mild increase of the endogenous diamine concentration (+8{\%}). HDC activity in both groups of diseased brains was significantly increased compared",
    author = "Gueli, {Maria Concetta}",
    year = "2013",
    language = "English",

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    T1 - High turnover rate of central histaminergic system in patients with Down syndrome and Alzheimer disease

    AU - Gueli, Maria Concetta

    PY - 2013

    Y1 - 2013

    N2 - HIGH TURNOVER RATE OF CENTRAL HISTAMINERGIC SYSTEM IN PATIENTS WITH DOWN SYNDROME AND ALZHEIMER DISEASE Maria Concetta Gueli Dipartimento di Biomedicina Sperimentale e Neuroscienze Cliniche (BioNEC), Università degli Studi di Palermo It is well confirmed that a strong relationship exists between Down’s syndrome (DS) and Alzheimer’s disease (AD). Neurochemical investigations reported that many central neurotransmitter systems are similarly affected in aging Down and in Alzheimer patients, respectively. Airaksinem et al. (1) found numerous neurofibrillary tangles in the tuberomammillary area of the hypothalamus, where cell bodies of histaminergic neurons are located. While Mazurkiewicz-Kwilecki et al. (2) found deficits of the endogenous diamine, Cacabelos et al. (3) reported an increase of central histamine levels. In the present study, in order to test whether AD-like neuropathological changes involve the central histaminergic system, we measured the concentration of histamine, histidine as well as the activity of histidine decarboxylase (HDC) and histamine-Nmethyltransferase (HMT) in temporal cortex (TC) of aging Down, Alzheimer and control patients. Post-mortem samples (temporal cortex, TC; grey matter) of AD neuropathologically confirmed cases (72.1 ± 7.6 years old), of karyotyped patients with DS (56.1 ± 7.1 years old), and control adults (72,7 ± 9.7 years old) were obtained from the MRC London Brain Bank for Neurodegenerative Diseases, Department of Neuropathology, Institute of psychiatry, London, U.K. Each block of brain tissue from AD, DS and controls were thawed on ice and homogenized in ice-cold HDC-solution of 0.1 M sodium phosphate buffer (pH 6.8) containing dithiothreitol and antipain protease inhibitor. Homogenates of brain specimens were centrifuged at 12,000 x g for 20 min at 4°C. The supernatants were poured into CENTRIPEP-3 concentrators (Amicon), and centrifuged at 2,000 x g for two 10 min periods at 4°C. The clear extracts were stored in small quantities in Eppendorf tubes at -80°C until analysis. HDC activity has been measured with the procedure described by Gueli et al. (4) and briefly summarized. Extract aliquots were pre-incubated for 10 min with HDC assay-solution (0.1 M PBS, 0.2 mM DTT, 0.01 mM PLP, 0.1 mM Aminoguanidine), then incubation was started by adding 0.5 mM Lhistidine for 0-3 h at 37°C. At the established times, the reactions were stopped with 60 % ice-cold PCA, and stored overnight. Finally, the reaction mixture was centrifuged at 19,000 x g for 30 min at 4°C. The supernatants were withdrawn and filtered (0.45 mm Millipore filter). The HPLC system consisted of a 600E Waters pump with a Waters 474 scanning fluorescence detector (ex 350 nm, em 450 nm). Chromatograms and calculations were performed by Empower TM2 Data Software. Histamine was separated and quantified after pre-column derivatization with Shore’s o-phthalaldehyde reaction (5), using a Spherisorb ODS2 analytical column, particle size 3 mm (20 x 0.46 cm,) (Waters, Milano), a 10 L injection volume, and a mobile phase of methanol, 20 mmol/ L sodium acetate in water, acetic acid (55:43:2 v/v) and 0.33 mmol/L 1- octanesulfonic acid sodium salt. The flow rate was 1.0 ml/min. In order to measure HMT activity brain tissue was disperged with a glass Teflon homogenizer in 0.1 M PBS (pH 7.2). After centrifugation the supernatant was used for the radioenzymatic assay (6). Histidine contents were measured using the procedure described by Borum (7). We observed a increase of histamine levels in temporal cortex of AD (+15%) patients. Down brains also showed a mild increase of the endogenous diamine concentration (+8%). HDC activity in both groups of diseased brains was significantly increased compared

    AB - HIGH TURNOVER RATE OF CENTRAL HISTAMINERGIC SYSTEM IN PATIENTS WITH DOWN SYNDROME AND ALZHEIMER DISEASE Maria Concetta Gueli Dipartimento di Biomedicina Sperimentale e Neuroscienze Cliniche (BioNEC), Università degli Studi di Palermo It is well confirmed that a strong relationship exists between Down’s syndrome (DS) and Alzheimer’s disease (AD). Neurochemical investigations reported that many central neurotransmitter systems are similarly affected in aging Down and in Alzheimer patients, respectively. Airaksinem et al. (1) found numerous neurofibrillary tangles in the tuberomammillary area of the hypothalamus, where cell bodies of histaminergic neurons are located. While Mazurkiewicz-Kwilecki et al. (2) found deficits of the endogenous diamine, Cacabelos et al. (3) reported an increase of central histamine levels. In the present study, in order to test whether AD-like neuropathological changes involve the central histaminergic system, we measured the concentration of histamine, histidine as well as the activity of histidine decarboxylase (HDC) and histamine-Nmethyltransferase (HMT) in temporal cortex (TC) of aging Down, Alzheimer and control patients. Post-mortem samples (temporal cortex, TC; grey matter) of AD neuropathologically confirmed cases (72.1 ± 7.6 years old), of karyotyped patients with DS (56.1 ± 7.1 years old), and control adults (72,7 ± 9.7 years old) were obtained from the MRC London Brain Bank for Neurodegenerative Diseases, Department of Neuropathology, Institute of psychiatry, London, U.K. Each block of brain tissue from AD, DS and controls were thawed on ice and homogenized in ice-cold HDC-solution of 0.1 M sodium phosphate buffer (pH 6.8) containing dithiothreitol and antipain protease inhibitor. Homogenates of brain specimens were centrifuged at 12,000 x g for 20 min at 4°C. The supernatants were poured into CENTRIPEP-3 concentrators (Amicon), and centrifuged at 2,000 x g for two 10 min periods at 4°C. The clear extracts were stored in small quantities in Eppendorf tubes at -80°C until analysis. HDC activity has been measured with the procedure described by Gueli et al. (4) and briefly summarized. Extract aliquots were pre-incubated for 10 min with HDC assay-solution (0.1 M PBS, 0.2 mM DTT, 0.01 mM PLP, 0.1 mM Aminoguanidine), then incubation was started by adding 0.5 mM Lhistidine for 0-3 h at 37°C. At the established times, the reactions were stopped with 60 % ice-cold PCA, and stored overnight. Finally, the reaction mixture was centrifuged at 19,000 x g for 30 min at 4°C. The supernatants were withdrawn and filtered (0.45 mm Millipore filter). The HPLC system consisted of a 600E Waters pump with a Waters 474 scanning fluorescence detector (ex 350 nm, em 450 nm). Chromatograms and calculations were performed by Empower TM2 Data Software. Histamine was separated and quantified after pre-column derivatization with Shore’s o-phthalaldehyde reaction (5), using a Spherisorb ODS2 analytical column, particle size 3 mm (20 x 0.46 cm,) (Waters, Milano), a 10 L injection volume, and a mobile phase of methanol, 20 mmol/ L sodium acetate in water, acetic acid (55:43:2 v/v) and 0.33 mmol/L 1- octanesulfonic acid sodium salt. The flow rate was 1.0 ml/min. In order to measure HMT activity brain tissue was disperged with a glass Teflon homogenizer in 0.1 M PBS (pH 7.2). After centrifugation the supernatant was used for the radioenzymatic assay (6). Histidine contents were measured using the procedure described by Borum (7). We observed a increase of histamine levels in temporal cortex of AD (+15%) patients. Down brains also showed a mild increase of the endogenous diamine concentration (+8%). HDC activity in both groups of diseased brains was significantly increased compared

    UR - http://hdl.handle.net/10447/104444

    M3 - Paper

    ER -