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Biomedical subjects

M Brasseur

Publications and source records attributed to M Brasseur.

At least 19 recordsLinked to original sources
PubMed1997

A high-throughput STAT binding assay using fluorescence polarization.

STAT (signal transducers and activators of transcription) is a class of transcription factors that are activated upon cytokine or growth factor binding to cell surface receptors. Activated STAT proteins dimerize, translocate into the nucleus, and activate transcription, leading to various immune responses. The inhibition of STAT binding to cell receptors might provide a means to modulate these immune responses. We developed a high-throughput biochemical assay to measure the interaction between an interferon-gamma receptor-derived phosphotyrosine-containing peptide and STAT1, using fluorescence polarization as the detection method. This assay can be used to screen for small molecules capable of disrupting receptor-STAT interactions.

Binding, Competitive↗
PubMed1997

A chemiluminescent microtiter plate assay for sensitive detection of protein kinase activity.

A chemiluminescent protein kinase assay using biotinylated substrate peptides captured on a streptavidin-coated microtiter plate and monoclonal antibodies to detect their phosphorylation is described. Assay conditions were optimized and validated for sensitive measurement of protein kinase A, protein kinase C, Ca2+/calmodulin-dependent protein kinase II (CAM-KII), receptor interacting protein, and src activities. The newly developed chemiluminescent assay has several advantages over currently used radioactive or colorimetric methods. It is highly sensitive at low enzyme and substrate concentrations and high, close to physiological ATP levels. It is fast, simple to perform and amenable to automation and high-throughput drug screening. The assay is also robust, exhibiting minimum interference from solvents and test substances from various sources. Overall, among the presently available methods for the detection of protein kinase activity, chemiluminescence was found to provide the highest sensitivity under conditions most closely mimicking the intracellular environment. This assay is expected to be useful in both academic and industrial laboratories, especially in identifying novel classes of protein kinase inhibitors.

Adenosine Triphosphate↗
PubMed1997

Some aspects of the acute phase response after a marathon race, and the effects of glutamine supplementation.

Strenuous exercise may be associated with immune suppression. However, the underlying mechanism is not known. A decrease in the plasma level of glutamine, which is utilised at a high rate by cells of the immune system, and an increase in the plasma level of some cytokines may impair immune functions such as lymphocyte proliferation after prolonged, exhaustive exercise. In two separate studies of the Brussels marathon, using similar protocols, the time course of the changes in the plasma concentrations of some amino acids (glutamine, glutamate, alanine, tryptophan and branched chain amino acids), acute phase proteins and cytokines (interleukins IL-1 alpha, IL-2, IL-6, tumour necrosis factor type a) was measured in male athletes. The numbers of circulating leucocytes and lymphocytes were also measured. Amino acid and cytokine concentrations have not previously been measured concomitantly in marathon runners; the measurement of some of these parameters the morning after the marathon (16 h) is novel. Another novel feature is the provision of glutamine versus placebo to marathon runners participating in the second study. In both studies the plasma concentrations of glutamine, alanine and branched chain amino acids were decreased immediately after and 1 h after the marathon. Plasma concentrations of all amino acids returned to pre-exercise levels by 16 h after exercise. The plasma concentration of the complement anaphylotoxin C5a increased to abnormal levels after the marathon, presumably due to tissue damage activating the complement system. There was also an increase in plasma C-reactive protein 16 h after the marathon. The plasma levels of IL-1 alpha were unaffected by the exercise, while that of IL-2 was increased 16 h after exercise. Plasma IL-6 was increased markedly (approximately 45-fold) immediately after and at 1 h after exercise. Neopterine, a macrophage activation marker, was significantly increased post-exercise. There was a marked leucocytosis immediately after the marathon, which returned to normal 16 h later. At the same time there was a decrease in the number of T-lymphocytes, which was further reduced within 1 h to below pre-exercise levels. Glutamine supplementation, as administered in the second study, did not appear to have an effect upon lymphocyte distribution.

Acute-Phase Proteins↗
PubMed1995

Components of a Stat recognition code: evidence for two layers of molecular selectivity.

Latent and activated forms of Stat1 and Stat6 have been expressed and purified, enabling biochemical experiments relating to their functional activities. Stat1 bound to a phosphotyrosine peptide derived from the IFN gamma receptor with a KD of 50 nM, whereas Stat6 bound to an IL-4 receptor peptide with a KD of 300 nM. Stat-receptor peptide interactions were specific and dependent upon tyrosine phosphorylation. Activated forms of Stat1 and Stat6 were used to select their optimal DNA binding sites. Stat1 selected a recognition site having dyad half-sites separated by 3 bp. Stat6 selected a recognition site composed of the same dyad half-sites, yet separated by 4 bp. Chimeric Stat1-Stat6 recombinants were expressed, purified, and assayed for receptor coupling and DNA binding specificity. Such studies led to the identification of polypeptide domains that specify these activities. These observations provide a framework for understanding how different cytokines elicit distinctive patterns of gene expression.

Amino Acid Sequence↗
PubMed1994

An interleukin-4-induced transcription factor: IL-4 Stat.

Interleukin-4 (IL-4) is an immunomodulatory cytokine secreted by activated T lymphocytes, basophils, and mast cells. It plays an important role in modulating the balance of T helper (Th) cell subsets, favoring expansion of the Th2 lineage relative to Th1. Imbalance of these T lymphocyte subsets has been implicated in immunological diseases including allergy, inflammation, and autoimmune disease. IL-4 may mediate its biological effects, at least in part, by activating a tyrosine-phosphorylated DNA binding protein. This protein has now been purified and its encoding gene cloned. Examination of the primary amino acid sequence of this protein indicates that it is a member of the signal transducers and activators of transcription (Stat) family of DNA binding proteins, hereby designated IL-4 Stat. Study of the inhibitory activities of phosphotyrosine-containing peptides derived from the intracellular domain of the IL-4 receptor provided evidence for direct coupling of receptor and transcription factor during the IL-4 Stat activation cycle. Such observations indicate that IL-4 Stat has the same functional domain for both receptor coupling and dimerization.

Amino Acid Sequence↗
PubMed1994

The 24-hour profiles of cortisol, prolactin, and growth hormone secretion in mania.

OBJECTIVE: To characterize sleep and the 24-hour profiles of cortisol, prolactin (PRL), and growth hormone (GH) secretion in mania. METHODS: Blood was sampled at 15-minute intervals, and sleep was polygraphically recorded in eight unmedicated male patients with pure mania and the results compared with those from a group of 14 healthy age-matched controls. The circadian, sleep-related, and pulsatile hormonal variations were quantitatively characterized using specifically designed computer algorithms. RESULTS: The manic state was associated with alterations of corticotropic activity and circadian rhythmicity partially overlapping those previously observed in acute endogenous depression, consisting of an elevation of nocturnal cortisol levels and an early timing of the nadir of the circadian variation. Sleep onset was delayed and the sleep period was reduced. A trend for short rapid eye movement latencies was apparent in the adult patients. Both the amount and the temporal organization of PRL and GH secretion were normal. CONCLUSION: The manic state seems to be characterized by similar but less severe neuroendocrine and circadian abnormalities, compared with major depression.

Acute Disease↗
PubMed1992

Reversal of cold induced haemoconcentration.

Classically, cold induced plasma volume reduction is explained by an increased diuresis which is generated by an inhibition of antidiuretic hormone release. However, most of the haemoconcentration appears to be reversible during rewarming. This observation weakens the former statement. The aim of this study was to clarify the mechanisms involved in the reversal of the cold induced haemoconcentration. Six young males, resting in a dorsal reclining position, were exposed successively to a thermoneutral environment (30 min), a cold environment (1 degrees C; cold) or thermoneutrality (control) for 120 min, and during a 60-min recovery period in thermoneutral conditions. During cold stress, a reduction of 15% (i.e. 510 ml) of the plasma volume was observed, and osmolality was unchanged. After the 60-min recovery under thermoneutral conditions, plasma volume variation between the Cold and the Control experiments was reduced and reached 3% (i.e. 100 ml). This volume equalled the increased amount of urine production observed during the cold stress experiment. Haemoconcentration cannot be explained by increased urinary water loss (+/- 100 ml) alone. Therefore a transient shift of plasma water from vascular to interstitial spaces, due to an increase of blood pressure, could be involved in the reduction of plasma volume.

Adult↗
PubMed1991

Tracheal antimicrobial peptide, a cysteine-rich peptide from mammalian tracheal mucosa: peptide isolation and cloning of a cDNA.

Extracts of the bovine tracheal mucosa have an abundant peptide with potent antimicrobial activity. The 38-amino acid peptide, which we have named tracheal antimicrobial peptide (TAP), was isolated by a sequential use of size-exclusion, ion-exchange, and reverse-phase chromatographic fractionations using antimicrobial activity as a functional assay. The yield was approximately 2 micrograms/g of wet mucosa. The complete peptide sequence was determined by a combination of peptide and cDNA analysis. The amino acid sequence of TAP is H-Asn-Pro-Val-Ser-Cys-Val-Arg-Asn-Lys-Gly-Ile-Cys-Val-Pro-Ile-Arg-Cys-Pr o- Gly-Ser-Met-Lys-Gln-Ile-Gly-Thr-Cys-Val-Gly-Arg-Ala-Val-Lys-Cys-Cys-Arg- Lys-Lys - OH. Mass spectral analysis of the isolated peptide was consistent with this sequence and indicated the participation of six cysteine residues in the formation of intramolecular disulfide bonds. The size, basic charge, and presence of three intramolecular disulfide bonds is similar to, but clearly distinct from, the defensins, a well-characterized class of antimicrobial peptides from mammalian circulating phagocytic cells. The putative TAP precursor is predicted to be relatively small (64 amino acids), and the mature peptide resides at the extreme carboxyl terminus and is bracketed by a short putative propeptide region and an inframe stop codon. The mRNA encoding this peptide is more abundant in the respiratory mucosa than in whole lung tissue. The purified peptide had antibacterial activity in vitro against Escherichia coli, Staphylococcus aureus, Klebsiella pneumonia, and Pseudomonas aeruginosa. In addition, the peptide was active against Candida albicans, indicating a broad spectrum of activity. This peptide appears to be, based on structure and activity, a member of a group of cysteine-rich, cationic, antimicrobial peptides found in animals, insects, and plants. The isolation of TAP from the mammalian respiratory mucosa may provide insight into our understanding of host defense of this vital tissue.

Amino Acid Sequence↗
PubMed1991

Circadian and sleep-related endocrine rhythms in schizophrenia.

Plasma levels of prolactin, growth hormone, corticotropin, and cortisol were measured at 15-minute intervals for 24 hours in nine unmedicated male schizophrenic patients and in nine age-matched normal male subjects. Each study was preceded by 3 days of habituation to the laboratory environment. Sleep was polygraphically recorded. The circadian and pulsatile variations present in each hormonal profile were quantitatively characterized with the use of computer algorithms specifically designed for analyses of hormonal fluctuations. The major abnormality of neuroendocrine release that was observed in the schizophrenic patients was an almost threefold enhancement of the sleep-related increase in the prolactin level, associated with an intensified frequency of nocturnal prolactin pulses. This increased stimulatory effect of sleep on prolactin secretion was evident immediately after sleep onset. The normal inhibition of cortisol secretion during early sleep was absent in schizophrenic patients. The major sleep abnormalities were a prolonged sleep latency and a reduction in total rapid eye movement stage sleep. During wakefulness, prolactin and cortisol levels were normal. The 24-hour profile of growth hormone was unaltered in schizophrenic patients, and a sleep-onset growth hormone pulse was observed in all patients. No abnormalities were noted in the levels or temporal organization of corticotropin secretion. Both the amplitude and the timing of the cortisol rhythm were normal. We conclude that, in schizophrenic men, pituitary-adrenal function and circadian time-keeping are normal but prolactin secretion is hyperresponsive to the physiologic stimulus of sleep onset. Schizophrenia thus appears to be characterized by a subset of neuroendocrine disturbances distinct from that observed in major endogenous depression.

Adrenocorticotropic Hormone↗
PubMed1990

Hematological variations at rest and during maximal and submaximal exercise in a cold (0 degree C) environment.

The affect of negative thermal stress on hematological variables at rest, and during submaximal (sub ex) and maximal exercise (max ex) were observed for young males who volunteered in two experimental sessions, performed in cold (0 degree C) and in normal room temperature (20 degrees C). At rest, hematological variables such as RBC and derivates Hb and Hct were significantly increased (P less than 0.05) during cold stress exposure, while plasma volume decreased. The findings of this study suggest that the major factor inducing hypovolemia during low thermal stress can be imputed to local plasma water-shift mechanisms and especially to a transient shift of plasma water from intra- to extravascular compartments. Rest values for WBC and platelets (Pla) were also slightly increased during cold stress exposure. However this increase can partly be related to hemoconcentration but also to the cold induced hyperventilation activating the lung circulation. Maximal exhaustive exercise induced, in both experimental temperatures, significant (P less than 0.05) increments of RBC, Hb, Hct, and WBC while plasma volume decreased. However, Pla increase was less marked. On the other hand, cold stress raised slightly the observed variations of the different hematological variables. Submaximal exercise induced a similar, though non-significant, pattern for the different hematological variables in both experimental conditions. Observed plasma volume (delta PV%) reduction appears during exercise. However cold stress induced resting plasma volume variations that are transferred at every exercise level. Neither exercise nor cold inducement significantly modified the hematological indices (MCH, MCV, MCHC). In conclusion hematological variables are affected by cold stress exposure, even when subjects perform a physical activity.

Acclimatization↗
PubMed1988

[Neuroendocrine rhythms in uni- and bipolar depressions].

Eighteen patients with major depressive disorder of the endogenous subtype (8 unipolars and 10 bipolars) were submitted to blood sampling at 15 min interval for 24 h with polygraphic sleep recording during an acute episode of depression. Plasma growth hormone (GH), adrenocorticotrophin (ACTH) and cortisol were measured in each sample. The depressed patients hypersecreted GH during the daytime and had hypercortisolism which was evident throughout the 24 h span. The nadir of ACTH and cortisol rhythms was advanced by an average of 3 h as compared to the timing observed in normal subjects. These abnormalities were more pronounced and more consistent in patients with unipolar rather than bipolar depression. These results are consistent with the hypothesis that disorders of circadian time-keeping may characterize major endogenous depression.

Adrenal Cortex Hormones↗
PubMed1988

Fifty cases of human immunodeficiency virus (HIV) infection: immunoultrastructural study of circulating lymphocytes.

The peripheral lymphocytes of 50 cases of human immunodeficiency virus (HIV) infection (13 of acquired immune deficiency syndrome (AIDS), 17 of AIDS related complex (ARC), and 20 healthy carriers) were studied immunoultrastructurally. The prevalence of "tubuloreticular structures" and "tubular confronting cisternae" increased with the progression of the disease. Numerous tubular confronting cisternae were noted in patients presenting with a high serum acid labile alpha-interferon values. The patients with depressed natural killer cell activity were characterised by circulating immature natural killer cells with abundant multivesicular bodies that were devoid of "parallel tubular arrays". With an immunogold staining technique the location of HIV antigen was detected ultrastructurally, both at the surface of "hand-mirror" natural killer cell lymphocytes and inside vacuolised cells, probably corresponding to infected T4 lymphocytes. These findings indicate the usefulness of electron microscopic techniques in evaluating the pathology and the pathogenetic outcome of AIDS.

AIDS-Related Complex↗
PubMed1988

[Hematological variations with submaximal long-term physical exercise].

The purpose of this study was to determine the effects of cold stress on routine hematologic parameters when subjects are submitted on long-lasting exercise. Eight male subjects, aged 22.7 +/- 1.3 years with a peak oxygen consumption of 54.3 +/- 5.2 mL/kg/min performed, respectively at 20 degrees C and 0 degrees C, a two-hour submaximal cyclergometry at an intensity of 40% of that performed during a previous exhaustive test. A 21-gauge catheter was inserted into an ante cubital vein for collection of the blood samples. Platelets and WBC count was significantly (p less than 0.05) increased during exercise in both 20 degrees C and 0 degrees C environmental temperatures. A slight but not statistically significant increase of RBC, Hb and Hct and a plasma concentration during cold exposure was observed. Indices are unchanged in both experiments. The findings of this study suggest that physical exercise is a major factor inducing hypervolemia and that low thermal environment does not appear to be an effective factor influencing hematologic variables during exercise.

Adaptation, Physiological↗
PubMed1987

24-hour profiles of adrenocorticotropin, cortisol, and growth hormone in major depressive illness: effect of antidepressant treatment.

Plasma ACTH, cortisol, and GH concentrations were measured at 15-min intervals for 24 h in 11 men suffering from major depressive illness during an acute episode of depression and during clinical remission following antidepressant treatment with either electroconvulsive therapy or amitriptyline. Seven age-matched normal men also were studied. During the acute phase of the illness, the patients had abnormally short rapid eye movement sleep latencies, hypercortisolism, early timing of the nadirs of the ACTH-cortisol rhythms, and shorter nocturnal periods of quiescent cortisol secretion. GH was hypersecreted during wakefulness, and a major pulse occurred before, rather than after, sleep onset. After treatment, rapid eye movement sleep latencies were lengthened, and cortisol levels returned to normal due to a decrease in the magnitude of episodic pulses. Moreover, the timing of the circadian rhythms of ACTH and cortisol as well as the duration of the quiescent period of cortisol secretion were normalized. The amount of GH secreted during wakefulness decreased to normal values, with fewer significant GH pulses. The major elevation of GH secretion in the early part of the night occurred later than that during the depressive episode. These results demonstrate that a disorder of circadian rhythmicity characterizes acute episodes of major depressive illness and that this chronobiological abnormality as well as the hypersecretion of ACTH, cortisol, and GH are state rather than trait dependent.

Adrenocorticotropic Hormone↗
PubMed1986

[Endurance bicycle ergometry at 0 degrees C: cardio-respiratory and metabolic course].

The purpose of the present study is to determine the physiological and metabolic variations during and after an exhaustive exercise in different thermal conditions (0 degree C and 20 degrees C). For the eleven experimental subjects, the maximal power output is the same at 0 degrees C and 20 degrees C: 330 +/- 30 W (Mean +/- s). The heart rate is lower (bradycardia) at rest, exhaustion, and recovery. However, these differences are only significant (p less than 0.05) at exhaustion. The oxygen consumption and pulmonary ventilation is higher at 0 degree C in comparison with the measurements at 20 degrees C. The blood lactate concentrations are significantly lower at 0 degree C and we observe the inverse phenomenon for the bicarbonates. The bases excess was significant at 4, 6 and 10th minute of recovery time. These values are higher for the measurements taken at 0 degree C. The results have shown that the cold stress after an exhaustive exercise and during recovery, can induce a physiological and metabolic response unlike of those at 20 degrees C.

Adolescent↗