Search PubMed⌕ Search

Biomedical subjects

R Ekman

Publications and source records attributed to R Ekman.

At least 55 records · Page 3Linked to original sources
PubMed1999

Identification of defensins in human lymphocyte nuclei.

The cell nucleus plays an essential role in all aspects of cell function and regulation. Most of the nuclear proteins/peptides are synthesized in the cytoplasm and transported into the nucleus through the nuclear pore complexes. The nuclear proteins/peptides conjugate with each other and interact in transcriptional activation/inactivation. Several of the high molecular mass transcription factors (> 30 kDa) have been identified and characterized. However, the information on the low molecular mass proteins/peptides of the nucleus is limited. We have investigated these low molecular mass proteins/peptides from the nucleus of human peripheral blood lymphocytes using reversed-phase high-performance liquid chromatography (RP-HPLC). The HPLC fractions were further analysed by matrix assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry, electrospray ionization time of flight (ESI-TOF) mass spectrometry and electrospray ionization fourier transform ion cyclotron resonance (ESI-FTICR) mass spectrometry for mass determination. Using this combination of mass spectrometry techniques and microsequence analysis, we have shown that human lymphocyte nuclei contain defensins, a mixture of human neutrophil granule peptide 1, 2 and 3.

Amino Acid Sequence↗
PubMed1999

Acute ischemic chest pain is not associated with increased calcitonin gene-related peptide (CGRP) levels in peripheral plasma nor in the coronary circulation.

Calcitonin gene-related peptide (CGRP) and substance P co-exist in capsaicin-sensitive primary sensory neurons and are released from the myocardium after activation of sensory nerve fibres as well as by ischemia in animals. This study was undertaken to try to clarify the potential involvement of immunoreactive (ir) CGRP in anginal pain and myocardial ischemia in humans. One clinical group (n = 87) and one experimental group (n = 14) were studied. The clinical group was admitted to a coronary care unit with suspected or definite acute myocardial infarction (AMI). The experimental group consisted of patients with severe angina pectoris (NYHA III-IV). This group was subjected to atrial pacing up to the appearance of angina pectoris. Mean irCGRP levels at admission for the clinical group with and without AMI showed no significant difference. Neither were any significant differences found in irCGRP concentrations between patients with pain as compared to those without pain or in the group who had had chest pain >30 min before hospital admission as compared to those with chest pain <30 min. Extraction ratios for lactate and irCGRP was calculated in the experimental group. No statistically significant covariance was found between irCGRP extraction ratio and lactate extraction ratio (r(xy) = -0.006) at the time of appearance of angina during atrial pacing. Despite the facts that CGRP may be liberated by a variety of physiological stimuli and may act as a potent vasodilator in the human vasculature, no evidence has been found in this study that CGRP release is increased as a consequence of ischemia or ischemic pain.

Aged↗
PubMed1998

Cerebrospinal fluid monoamines in Prader-Willi syndrome.

BACKGROUND: The behavioral phenotype of Prader-Willi syndrome (PWS) suggests hypothalamic dysfunction and altered neurotransmitter regulation. The purpose of this study was to examine whether there was any difference in the concentrations of monoamine metabolites in the cerebrospinal fluid (CSF) in PWS and non-PWS comparison cases. METHODS: The concentration of monoamine metabolites in CSF was determined in 13 children and adolescents with PWS diagnosed on clinical and genetic criteria. The concentrations were compared with those from 56 comparison cases in healthy and other contrast groups. RESULTS: The concentrations of dopamine and particularly serotonin metabolites were increased in the PWS group. The differences were most prominent for 5-hydroxyindoleacetic acid. The increased concentrations were found in all PWS cases independently of age, body mass index, and level of mental retardation. CONCLUSIONS: The findings implicate dysfunction of the serotonergic system and possibly also of the dopamine system in PWS individuals, and might help inform future psychopharmacologic studies.

Adolescent↗
PubMed1998

[Study and treatment of heart failure require new approaches. Natriuretic peptides can be safe and simple diagnostic and outcome measures].

Analysis of plasma natriuretic peptides and related propeptide fragments may be a cost-effective aid to diagnostic evaluation and treatment follow-up in cases of heart failure. In diagnostic potential such variables may constitute first-line measures of high negative predictive value, allowing further examination, e.g. by echocardiography, in cases where values are above the respective cut-off levels. However, in many cases evaluation of published reports is rendered difficult by their omission of information on such pre-analytical variables as blood sampling and storage, and drug therapy. Moreover, different analytical methods may yield widely divergent results. Thus, before such assays are introduced in general use, their long-term validity needs to be ensured, for instance by consistency in calibration, and measurements need to be made in representative series of unselected patients for the determination of appropriate cut-off levels.

Amino Acid Sequence↗
PubMed1998

Suppressed neuropeptide Y (NPY) mRNA in rat amygdala following restraint stress.

We have previously demonstrated that NPY produces anxiolytic-like effects through actions in the amygdala, and that anxiogenic-like effects of restraint stress are mediated through this structure. Here, we examined the effects of restraint stress on NPY mRNA levels in amygdala and several other brain regions. A sensitive solution hybridization-RNase protection assay (RPA) was developed, employing a combination of internal and external standards, which allowed absolute quantitation of NPY mRNA in tissue-samples of less than 10 mg. NPY mRNA levels were determined, following a 1-h restraint stress, in homogenates of tissue from the amygdala, neocortex, striatum and hypothalamus, and the time course of these effects was examined. A highly significant decrease in NPY-mRNA levels was seen in the amygdala at 1 h and 2 h following restraint, with levels returning to normal within 10 h. A similar effect was seen in the neocortex, but was less pronounced and slower in onset. Striatal and hypothalamic NPY expression was not significantly affected. Tissue levels of NPY-peptide were modestly decreased in the amygdala at 1 h following restraint and had returned to normal within 4 h. The present findings support the hypothesis that anxiety related behavioral effects of stress may in part be mediated through modulation of NPY function in the amygdala.

Amygdala↗
PubMed1998

High delta sleep-inducing peptide-like immunoreactivity in plasma in suicidal patients with major depressive disorder.

BACKGROUND: Delta sleep-inducing peptide (DSIP) supposedly involves the hypothalamus-pituitary-adrenal (HPA) axis. Previous studies have shown deviated plasma DSIP-like immunoreactivity (DSIP-LI) levels, as well as abnormal DSIP-LI responses to corticotropin-releasing hormone in patients with major depressive disorder (MDD). This study was performed to investigate plasma-DSIP-LI and its association with the dexamethasone suppression test (DST) in suicide attempters. METHODS: Plasma-DSIP and serum cortisol were measured before and after dexamethasone intake in 34 suicide attempters and in healthy age- and sex-matched controls. RESULTS: We found significantly elevated DSIP-LI levels in MDD patients (p < .005), and a significant correlation between predexamethasone cortisol and predexamethasone DSIP-LI levels in healthy controls. Postdexamethasone DSIP-LI levels increased in subjects with low predexamethasone DSIP-LI levels, whereas they decreased in subjects with high predexamethasone DSIP-LI levels. CONCLUSIONS: Results give some support to the theory of glucocorticoid involvement in the regulation of DSIP, and suggest altered DSIP-LI levels in suicidal MDD patients.

Adult↗
PubMed1998

Sensory CGRP depletion by capsaicin exacerbates hypoxia-induced pulmonary hypertension in rats.

Pulmonary hypertension is a debilitating disease that occurs among infants and adults. One of many etiologies is airway hypoxia. We previously demonstrated a role of endogenous calcitonin gene-related peptide (CGRP), a potent vasodilator, in ameliorating the pulmonary vascular pressor response to chronic hypoxia and related changes in the lungs and heart. This study evaluates the role of endogenous sensory CGRP in hypoxic pulmonary hypertension and examines the intrinsic neural microcircuitry. Rats were pretreated with capsaicin i.p. to deplete pulmonary sensory C-fiber stores of CGRP and substance P and placed in hypobaric hypoxia (10% O2, 16 days) or normoxia together with sham controls. Hypoxia increased pulmonary artery pressure, right-ventricular weight, arterial medial thickness, elasticized capillaries, endothelial cell density, lung water and hematocrit in control rats. Capsaicin augmented pulmonary artery pressure and right-ventricular hypertrophy in hypoxia, and medial thickness and endothelial cell density both in normoxia and hypoxia. Because of the limited effects on these parameters by substance P and other capsaicin-sensitive lung agents, our results suggest that sensory CGRP deficit severely exacerbates pathological signs of hypoxic pulmonary hypertension. A neural microcircuitry consistent with an axon reflex pathway is outlined histochemically. We conclude that endogenous CGRP modulates pulmonary vascular tone in hypoxic pulmonary hypertension which requires intact primary sensory fibers.

Animals↗
PubMed1998

Substance P and related peptides in porcine cortex: whole tissue and nuclear localization.

A large portion of the knowledge that has been gathered on the distribution of neuropeptides in neural tissues is based on findings obtained with immunocytochemistry and radioimmunoassay. However, these methods give limited structural information about the peptides being studied. Using porcine cortex as a model tissue, we combined immunoaffinity chromatography with reversed-phase high-performance liquid chromatography, radioimmunoassay, and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS). We determined the molecular nature of the peptides contributing to the substance P-like immunoreactivity measured in extracts of whole tissue and cell nuclei. In addition to substance P(1-11), other peptides were extracted using this protocol. The presence of SP(1-11) was confirmed through post-source decay analysis. These results illustrate the usefulness of MALDI-TOF-MS in the characterization of neuropeptides from biological tissues.

Amino Acid Sequence↗
PubMed1998

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis of proteins in human cerebrospinal fluid.

Matrix-assisted laser desorption/ionization time-of-flight mass spectra of proteins in cerebrospinal fluid analyzed without prior purification are presented. Less than 100 fmol amounts of proteins in the 10,000 to 20,000 u mass range and linked to human disease (multiple sclerosis, Alzheimer's disease, and stroke) were detected in a complex mixture of proteins and peptides, in the presence of high concentrations of salts, lipids and free amino acids. The mass resolution was sufficient to distinguish between the non-hydroxylated and hydroxylated forms of a 13,400 u protein. Simple fractionation of the cerebrospinal fluid using microbore-reversed phase high performance liquid chromatography improved signal-to-noise ratios in the mass spectra. High-accuracy peptide mass mapping and database searching were utilized to confirm the identity of several proteins. The presented results show that matrix-assisted laser desorption/ionization time-of-flight mass spectrometry could be used as a tool to perform rapid screening of chemically altered proteins in small volumes of biological fluids.

Blotting, Western↗
PubMed1998

Correlations between plasma-neuropeptides and temperament dimensions differ between suicidal patients and healthy controls.

BACKGROUND: Decreased plasma levels of plasma-neuropeptide Y (NPY) and plasma-corticotropin releasing hormone (CRH), and increased levels of plasma delta-sleep inducing peptide (DSIP) in suicide attempters with mood disorders have previously been observed. This study was performed in order to further understand the clinical relevance of these findings. METHODS: Examination of correlates between temperament dimensions (Karolinska Scales of Personality (KSP), the Eysenck Personality Questionnaire together with the IVE- impulsiveness scale (EPQI), and the Marke-Nyman Temperament (MNT)) and NPY, CRH and DSIP and serum-cortisol in the dexamethasone suppression test (DST) in 38 suicidal patients and matched controls. RESULTS: NPY correlated significantly and positively with psychasthenia, irritability, and stability and significantly and negatively with validity in patients, but significantly and negatively with muscular tension, psychasthenia, verbal aggression and irritability in controls. DSIP correlated significantly and positively with impulsiveness (EPQI) in controls. CRH correlated negatively with lie in controls. Cortisol correlated significantly and positively with validity, extraversion and verbal aggression and significantly and negatively with inhibition of aggression in controls. CONCLUSION: NPY may be related to stress tolerance. DSIP seems to be associated with impulsivity/antisocial traits. LIMITATIONS: Non-suicidal patients were not included in the examination. CLINICAL RELEVANCE: The state of depression or stress seems to influence the correlations studied.

Adult↗
PubMed1998

Mass spectrometry of peptides in neuroscience.

This review focuses on the contributions of modern mass spectrometry to neuropeptide research. An introduction to newer mass spectrometric techniques is provided. Also, the use of mass spectrometry in combination with high-resolution separation techniques for neuropeptide identification in biological samples is illustrated. The amino acid sequence information that is important for the identification and analysis of known, novel, or chemically modified neuropeptides may be obtained using mass spectrometric techniques. Because mass spectrometry techniques can be used to reflect the dynamic properties associated with neuropeptide processing in biological systems, they may be used in the future to monitor peptide profiles within organisms in response to environmental challenges such as disease and stress.

Mass Spectrometry↗
PubMed1998

Processing of neuropeptide Y and somatostatin in human cerebrospinal fluid as monitored by radioimmunoassay and mass spectrometry.

The processing of four neuropeptides, neuropeptide Y (NPY) 1-36, NPY (18-36), somatostatin (SOM) 1-28, and SOM (15-28) was studied in human cerebrospinal fluid (CSF) by using a novel combination of methods that included radioimmunoassay (RIA) and mass spectrometry. Untreated CSF samples were chromatographed using reversed-phase high pressure liquid chromatography (RP-HPLC) followed by NPY-RIA or SOM-RIA. These results were compared with those obtained by incubating CSF with exogenous synthetic peptides and directly detecting peptide fragments by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-MS). Using this combination of methods, we were able to determine the probable identities of peptides/peptide fragments recognized in radioimmunoassays. The most important NPY-immunoreactive components in CSF were found to be NPY (1-36) and NPY (3-36). Metabolic products of SOM (15-28) were found to contribute to SOM-like immunoreactivity (SOM-LI) in CSF, but SOM (1-28) only to a lesser degree. Differences in the rate of neuropeptide processing were observed. These differences depended more on the length of the peptide than its sequence. NPY (18-36) and SOM (15-28) were rapidly and extensively processed, whereas NPV (1-36) and SOM (1-28) were processed much more slowly in CSF. The production of SOM (15-28) from SOM (1-28) by enzymes in CSF was not observed. Also, the presence of a disulfide bond in the somatostatins appeared to stabilize them against enzymatic digestion of the ring structure. The results detailed in this report confirm MALDI-MS important role in studies of neuropeptide processing in CSF.

Amino Acids↗
PubMed1998

Heparin and vasoactive peptides in hemodialysis patients.

Plasma concentrations of vasoactive peptides have been reported to be influenced by various procedural features of hemodialysis (HD), such as ultrafiltration and isovolemic diffusion, dialysate buffer and dialysate temperature, but also by sham HD thus reflecting an effect of the extracorporeal circulation per se. In the present study the effect of heparin administration was investigated in 9 stable HD patients, and compared with that of saline. Blood samples were taken from the arteriovenous fistula before and 45 min after the administration of heparin or saline. After an interval of 2 weeks, the procedure was repeated with the exception that the patients who received heparin on the first occasion were given saline and vice versa. Plasma concentrations of the vasoactive peptides were measured by radioimmunoassay. Regardless of whether heparin was given or not, the plasma concentrations of the vasodilators atrial natriuretic peptide, beta-endorphin and vasoactive intestinal peptide did not change, nor did the concentration of the vasoconstrictor neuropeptide Y. The plasma motilin concentration decreased significantly when heparin was given, and that of substance P increased, both these peptides being vasodilators. Mean arterial blood pressure decreased regardless of whether heparin was given or not, and no difference between the two regimens was noted. Heart rate was unchanged with both regimens. To sum up, administration of heparin but not of saline affected the plasma concentrations of motilin and substance P. However, the decrease in blood pressure during the procedure seemed not to be related to the changes in these peptides, as it also occurred in the absence of heparin.

Aged↗
PubMed1998

Glucose stimulation of pancreatic beta-cell lines induces expression and secretion of dynorphin.

To investigate adaptive responses of pancreatic beta-cells to hyperglycemia, genes induced by glucose stimulation were identified by subtraction cloning. Among 53 clones representing differentially expressed genes, 20 encoded the endogenous opioid precursor, prodynorphin. The amino acid sequence of murine prodynorphin is identical to the rat protein in sequences comprising the opioid peptides and 86% identical in the remainder of the molecule. Stimulation of MIN6 cells increased prodynorphin RNA levels to more than 20-fold in proportion to physiological glucose concentrations. Similar induction levels were observed in murine betaTC3 and rat Rinm5F beta-cell lines. Prodynorphin RNA expression increased within 1 h of glucose stimulation, achieved maximal levels by 4 h, and remained elevated for at least 24 h. By using RIA, MIN6 cells were shown to contain and secrete increased amounts of dynorphin-A following glucose stimulation. Treatment of MIN6 cells with KCl, forskolin, or isobutyl-methyl-xanthine strongly induced prodynorphin RNA expression, suggesting that induction may be related to secretion-coupled signaling pathways. The induction of prodynorphin in several beta-cell lines is consistent with previous demonstrations of beta-cell synthesis of other endogenous opioids, including beta-endorphin, and suggests that opioids may have a potentially significant role in regulating beta-cell secretion.

Amino Acid Sequence↗
PubMed1997

Isolation and characterization of proteins from human lymphocyte nuclei using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and post-source decay analysis.

The cell nucleus plays an essential role in all aspects of cell function, including DNA replication, gene transcription, RNA processing and cell division. Within the cell nucleus there are many proteins and peptides that have regulatory roles. Although several higher molecular weight protein components (> 30 kDa) have been identified and characterized, limited information is available for the lower molecular weight components. Nuclei from human peripheral blood lymphocytes were isolated and rapidly characterized by employing a strategy using reversed-phase high-performance liquid chromatography, tryptic digestion, and post-source decay analysis of matrix-assisted laser desorption/ionization (MALDI) ions in combination with database searches. Database searches utilizing molecular weight, proteolytic digest fragments, and peptide sequence results identified known proteins. The results illustrate the usefulness of MALDI as a tool in the characterization of low abundance proteins in the cell nucleus.

Calibration↗
PubMed1997

Measurements of catecholamine-mediated apoptosis of immunocompetent cells by capillary electrophoresis.

Single cell analysis with capillary electrophoresis, a technique capable of detecting zeptomole quantities (10(-21) mole) of neurochemical species, has been used to demonstrate that lymphocytes are capable of active synthesis of dopamine and norepinephrine. Exposure of lymphocytes to catecholamines at concentrations as low as 10 nM leads to decreased proliferation and differentiation, e.g. interferon-gamma (IFN-gamma), interleukin-4 (IL-4) and immunoglobulin (Ig). In addition, both inhibition of dopamine uptake with nomifensine and inhibition of packing of catecholamines into vesicles with tetrabenazine, results in significantly lower levels of dopamine and norepinephrine (p < 0.01 and p < 0.05, respectively). The catecholamine-dependent inhibition of T- and B-lymphocyte activity is mediated via an induction of a Bcl-2/Bax and Fas/FasL involved apoptosis. These findings indicate a novel mechanism for regulation of lymphocyte activity in the central nervous system, whereby elevated regional levels of catecholamines might lead to the immunoprivilege of the brain.

Adrenergic Uptake Inhibitors↗
PubMed1997

A new procedure for detecting brain-specific proteins in cerebrospinal fluid.

We have developed a new procedure, including three affinity chromatography steps, micro-reversed phase high pressure liquid chromatography (mR-HPLC) and Western blotting/mass spectrometric analysis to study central nervous system (CNS) specific proteins in human cerebro spinal fluid (CSF) in order to find biochemical markers for neuronal and synaptic function and pathology in degenerative brain disorders. After the three affinity chromatography steps, intended to remove interfering serum proteins from CSF, mR-HPLC revealed four major peaks, which by both Western blotting and mass spectrometric analyses were found to correspond to beta 2-microglobulin, cystatin C, transthyretin (TTR) and asialotransferrin. When comparing these peaks in CSF from Alzheimer's disease (AD) patients and age-matched healthy controls, a reduction of the brain-specific TTR was found. Therefore we quantified TTR in CSF and serum samples from 8 patients with early onset AD (EAD), 18 patients with late onset AD (LAD), 8 patients with vascular dementia (VAD) and 18 healthy individuals using a nephelometric method. CSF-TTR was divided into barrier-dependent and barrier-independent TTR. The barrier-independent i.e. brain-specific TTR was significantly reduced in the EAD group compared to the controls. Transthyretin has been found to be present in the senile plaques in AD, and to specifically bind to beta/A4 protein, the major component of the amyloid deposits in AD. Therefore, the reduction of the transthyretin-isoform in CSF in AD may reflect an absorption of transthyretin to the amyloid deposits in the senile plaques.

Aged↗