Psychosocial stress-induced activation of salivary alpha-amylase: an indicator of sympathetic activity?
Female; Humans; Male; Adult; Middle Aged; Social Environment; Predictive Value of Tests; Stress; Biomarkers of Pain; Chromatography; High Pressure Liquid; Kinetics; Circadian Rhythm/physiology; Adrenal Medulla/physiopathology; alpha-Amylase/metabolism; Norepinephrine/metabolism; Psychological/enzymology/physiopathology; Saliva/enzymology; Sympathetic Nervous System/physiopathology
Assessment of sympathoadrenal medullary system (SAM) activity is only possible to date via measurement of catecholamines in blood plasma or via electrophysiological methods. Both ways of measurement are restricted to endocrinological or psychophysiological laboratories, as both require either immediate freezing of blood samples or complex recording devices. Efforts have therefore been undertaken to find a method comparable to salivary cortisol measurements, in which noninvasive samples can be taken at any place and stored at room temperature for sufficient time before later analysis in the laboratory. Salivary alpha-amylase (sAA) is a candidate that may prove useful in this context. We show here that sAA activity is increased by acute psychosocial stress (Trier Social Stress Test) and that increases in sAA correlate with increases in norepinephrine. We further report that sAA exhibits a stable circadian pattern that mirrors that of salivary cortisol. In conclusion, the current data show that salivary alpha-amylase may serve as an easy-to-use index for SAM activity. However, some questions remain to be answered; for example, what impact does salivary flow rate exert on stress-induced sAA activity?
2004
Rohleder N; Nater UM; Wolf JM; Ehlert U; Kirschbaum C
Annals Of The New York Academy Of Sciences
2004
Article information provided for research and reference use only. PedPalASCNET does not hold any rights over the resource listed here. All rights are retained by the journal listed under publisher and/or the creator(s).
Journal Article
<a href="http://doi.org/10.1196/annals.1314.033" target="_blank" rel="noreferrer">10.1196/annals.1314.033</a>
Quantitative analysis of methionine enkephalin and beta-endorphin in the pituitary by liquid secondary ion mass spectrometry and tandem mass spectrometry
Humans; Animals; Biomarkers of Pain; beta-Endorphin/analysis; Enkephalin; Methionine/analysis; Chromatography; High Pressure Liquid; Mass Spectrometry; Pituitary Gland/chemistry
This manuscript reviews the use of an off-line combination of liquid chromatography (LC) and mass spectrometry (MS) to quantify endogenous neuropeptides in biological tissues and fluids, and tandem MS (MS/MS) to optimize the molecular specificity of the quantification of native peptides. Reversed-phase high-performance liquid chromatography (RP-HPLC) was used to purify selected endogenous neuropeptides from biological tissues and fluids. Liquid secondary ion MS (LSI-MS), also known as fast atom bombardment (FAB), is used to desorb and to ionize the peptide. The corresponding stable isotope-incorporated synthetic peptide of each peptide is used as the internal standard (I.S.) for quantification. The measurement of methionine enkephalin (ME) and of beta-endorphin1-31 (BE) in the human pituitary is described. This analytical method offers the highest molecular specificity for the measurement of a fully post-translationally modified peptide.
1998
Desiderio DM; Zhu X
Journal Of Chromatography.A
1998
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Journal Article
<a href="http://doi.org/10.1016/s0021-9673(97)00670-5" target="_blank" rel="noreferrer">10.1016/s0021-9673(97)00670-5</a>
Novel opioid peptides endomorphin-1 and endomorphin-2 are present in mammalian immune tissues
Female; Humans; Male; Adult; Middle Aged; Animals; Rats; Non-U.S. Gov't; Research Support; Wistar; Radioimmunoassay/methods; Chromatography; High Pressure Liquid; Oligopeptides/analysis; Cross Reactions; Immune Sera/immunology; Immune System/chemistry; Spleen/chemistry; Thymus Gland/chemistry; Tissue Extracts/chemistry
Endomorphin (EM)-1 and EM-2 are opioid tetrapeptides, reported within the central nervous system, which have very high specificity and affinity for the mu-opioid receptor. We have used newly developed and well-characterised radioimmunoassays (RIAs) in combination with reversed-phase high-performance liquid chromatography (HPLC) to detect EM-1 and EM-2 immunoreactivity (ir) in rat immune tissues. Endomorphins were detectable in extracts of rat spleen (total EM-1-ir/spleen: 440+/-73 pg, mean+/-SEM, a=group of eight rats; EM-2-ir: 150+/-12 pg) and thymus (EM-1-ir: 152+/-18 pg, mean+/-SEM n=8; EM-2-ir: 156+/-28 pg). EM-2-ir was detectable in extracts of human spleen (338+/-196 pg/g tissue, n=3). Multiple peaks of EM-1-ir and EM-2-ir were observed in rat spleen and thymus extracts, and multiple peaks of EM-2-ir were observed in extracts of human spleen, following reversed-phase HPLC and RIAs. This is the first report of endomorphin immunoreactivity in tissues of the rat and human immune systems.
2000
Jessop DS; Major GN; Coventry TL; Kaye SJ; Fulford AJ; Harbuz MS; De Bree FM
Journal Of Neuroimmunology
2000
Article information provided for research and reference use only. PedPalASCNET does not hold any rights over the resource listed here. All rights are retained by the journal listed under publisher and/or the creator(s).
Journal Article
<a href="http://doi.org/10.1016/s0165-5728(99)00216-7" target="_blank" rel="noreferrer">10.1016/s0165-5728(99)00216-7</a>
Analysis of proopiomelanocortin (POMC) messenger ribonucleic acid and POMC-derived peptides in human peripheral blood mononuclear cells: no evidence for a lymphocyte-derived POMC system
Humans; Molecular Sequence Data; Biomarkers of Pain; RNA; Base Sequence; beta-Endorphin/analysis; Biomarkers Reference List; Leukocytes; Lymphocytes/metabolism; Polymerase Chain Reaction; Chromatography; High Pressure Liquid; Pro-Opiomelanocortin/genetics; Mononuclear/metabolism; Blotting; Cathepsin D/metabolism; DNA Probes; Gel; Messenger/blood; Northern; Peptide Fragments/analysis; Peptide Mapping; Southern
A number of recent studies suggest that cells of the immune system, e.g. peripheral blood mononuclear cells (PBMC), can synthesize and process POMC and secrete POMC-derived peptides, such as ACTH and endorphins, upon immune and hormonal challenges. From this, it has been proposed that POMC-derived peptides originating from lymphoid cells can function as hormones, for instance in a lymphoid-adrenal axis. In view of the important physiological implications of this proposal, the present study was designed to investigate the expression of the POMC gene in human PBMC and the production by these cells of alpha-, beta-, and gamma-endorphins (alpha E, beta E, and gamma E) peptides that are established end products of the posttranslational processing of POMC. PBMC of individual donors were used uncultured (fresh cells) or cultured for 24 and 48 h in the presence and absence of Concanavalin-A (Con-A), bacterial lipopolysaccharide, phytohemagglutinin, or CRH, and vasopressin, conditions that reportedly stimulate POMC activity in those cells, to investigate the presence of POMC transcripts by analysis of total RNA with Northern blotting and the reverse transcriptase polymerase chain reaction (RT-PCR). Large scale preparations containing over 10(9) cells (fresh, cultured with and without Con-A) originating from several donors were examined for the presence of POMC transcripts by analysis of poly(A)+ RNA on Northern blots and for the presence of alpha E, beta E, and gamma E by gel filtration over Sephadex G-75 and reverse phase HPLC, followed by assay of the fractions in four endorphin RIA systems with different specificities. On the Northern blots of total RNA, no POMC transcripts were detectable. In poly(A)+ RNA preparations, no full-length POMC mRNA was found, and it was estimated that the concentration of POMC mRNA, if present, was below approximately 0.005 transcript/cell in Con-A-stimulated cells and still lower in unstimulated cells. In accord with literature data, an 800- to 900-nucleotide POMC transcript was detected in cultured PBMC, and the levels of this transcript were stimulated by Con-A. In all samples analyzed with RT-PCR, a transcript spanning most of exons 2 and 3 was detectable only on Southern blots of the RT-PCR product, but not on agarose gels stained with ethidium bromide. Chromatographic analysis of endorphin immunoreactivities in cell extracts revealed no qualitative differences between the immunoreactive profiles of fresh PBMC or PBMC cultured with or without Con-A.(ABSTRACT TRUNCATED AT 400 WORDS)
1993
van Woudenberg AD; Metzelaar MJ; van der Kleij AA; de Wied D; Burbach JP; Wiegant VM
Endocrinology
1993
Article information provided for research and reference use only. PedPalASCNET does not hold any rights over the resource listed here. All rights are retained by the journal listed under publisher and/or the creator(s).
Journal Article
<a href="http://doi.org/10.1210/endo.133.5.8404638" target="_blank" rel="noreferrer">10.1210/endo.133.5.8404638</a>