Biomedical subjects

M Lacroix

Publications and source records attributed to M Lacroix.

At least 73 records · Page 4Linked to original sources

PubMed1996

Effects of secretory products of breast cancer cells on osteoblast-like cells.

The pathogenesis of breast cancer-induced osteolysis remains largely unknown. To evaluate the potential role of osteoblasts as target cells during this process, we incubated SaOS-2 human osteoblast-like cells (OBL) with culture media conditioned by proliferative (PM, 'Proliferation Media') or confluent (CfM, 'Confluence Media') MCF-7 human breast cancer cells. CfM decreased the growth of OBL by 26% (P < 0.01) while PM was without significant effect on this parameter. In contrast, both PM and CfM obtained from MCF-7 cultures increased the cyclic AMP (cAMP) response of OBL to the osteolytic agents PTH (10(-8) M) and PTH-related peptide (PTHrP, 10(-8) M) by a factor of about 3 (P < 0.001), and to prostaglandin E(2) (PGE(2),10(-6) M) by a factor of about 2 (P < 0.01). No significant modulation of OBL growth or sensitivity to PTH, PTHrP, or PGE2 was induced by media obtained from HBL-100 non-malignant immortalized breast epithelial cell cultures. 17betaestradiol (E(2), 10(-8) M) and the antiestrogen tamoxifen (Tam, 10(-7) M) added for 48 h to MCF-7 cultures before collecting conditioned media attenuated and potentiated, respectively, the PM- but not the CfM-induced increase in the response of OBL to PTH or PTHrP Along the same line, the addition to MCF-7 conditioned media of a polyclonal anti-transforming growth factor-beta (TGF-beta) antibody attenuated by about 25% (P < 0.01) the PM-induced increase in OBL response to PTH and PTHrP while abrogating the modulatory effects of E(2) and Tam on that response. Together, our results indicate that MCF-7 breast cancer cells secrete factors which inhibit the growth of OBL and increase their sensitivity to various osteolytic agents. TGF-beta was only partly responsible for these effects, and accounts for their modulation by E(2) and Tam. The identification of other osteoblast-modulatory factor(s) should contribute to a better understanding and treatment of breast cancer-induced osteolysis.

Adenocarcinoma↗

PubMed1996

Cauda equina histiocytosis X.

A young child was admitted with a cauda equina syndrome believed to be progressive, but unrelated to a congenital anomaly. Magnetic resonance imaging studies revealed the presence of an intradural tumor in the cauda equina. A yellow pasty tumor with nerve root infiltration was identified at surgery. Histologic features were pathognomonic for histiocytosis X. The patient's management and review of the literature are discussed.

Cauda Equina↗

PubMed1996

Characterization of the HLA-restrictive elements of a rubella virus-specific cytotoxic T cell clone: influence of HLA-DR4 beta chain residue 74 polymorphism on antigenic peptide-T cell interaction.

The influence of glutamic acid (E)-alanine (A) dimorphism at position 74 of the DR4 beta chain on cytotoxic T cell recognition of an antigenic rubella virus peptide, E1(273-284), was studied using a panel of B cell lines and B cell transfectants expressing different HLA-DRB1 alleles as antigen-presenting cells and targets in 51Cr-release assays. Only B cell lines expressing the DRB1*0403, DRB1*0406 or DRB1*0407 subtypes which shared a residue, E, at position 74 in the DR4 beta chain when sensitized with E1(273-284) elicited strong cytotoxic T lymphocyte responses. However, in direct binding and antibody inhibition assays, it was shown that biotinylated E1(272-285) could bind to DR molecules with residues other than E at position 74, including DRB1*0401, DRB1*0404 and DRB1*1101 expressed on transfectants. E1(272-285) bound with similar affinity to the transfectant with DRB1*0403, which has E at position 74, as well as the transfectant with DRB1*0404, which does not. When T-B cell engagement rates were compared in cell conjugate assays, the percentage of T-B conjugates was higher when peptide-pulsed transfectants with DRB1*0403 were used than with transfectants expressing DRB1*0404. Hence, the HLA DR beta 1 polymorphism at position 74, while not critical for the binding affinity of E1(272-285) to the HLA molecule, appears to be a primary determinant of restricted recognition and subsequent activation of the peptide-specific T cells.

Amino Acid Sequence↗

PubMed1996

Rubella reimmunization: comparative analysis of the immunoglobulin G response to rubella virus vaccine in previously seronegative and seropositive individuals.

Rubella virus (RV)-specific immunoglobulin G (IgG) antibodies were studied in military recruits undergoing unselected immunization with live attenuated measles, mumps, and rubella virus (MMR) vaccine. Three different whole-RV enzyme immunoassays (EIAs) and an epitope-specific EIA with a synthetic peptide (BCH-178c) representing a heutralization domain on the RV E1 envelope protein were used. Before vaccination, 84.2, 87.7, and 84.5% of the subjects tested (n = 399) were found to be seropositive (> 10 IU/ml or assay equivalent) by the three whole-RV EIAs, respectively, while only 82.5% were seropositive by the BCH-178c EIA. Although prevaccination seropositivity rates were similar for the whole-RV EIAs (sensitivity, 94 to 100%), many sera considered seropositive by the whole-RV EIAs had E1 peptide EIA antibody levels of < 10 IU/ml (sensitivity, 77.4 to 80.7%). One month after vaccination, 97.8, 97.2, and 93.5% of the subjects who were followed (n = 356) were seropositive by the three whole-RV EIAs, respectively, while 89% had BCH-178c peptide-specific IgG titers of > 10 IU/ml. After vaccination, depending on the assay used, up to 20.6% of initially seropositive individuals exhibited a greater than fourfold increase in RV-specific IgG, while up to 47.3% showed a greater than twofold increase. Increased antibody titers after vaccination (seroboosting) were most frequently associated with low levels of BCH-178c peptide-specific IgG before vaccination. RV protein-specific IgG was also studied by immunoblot assays in a subset (n = 56) of individuals receiving the MMR vaccine. Of these, 89.4 and 91.1% exhibited RV protein (E1, E2, and C protein)-specific IgG before and after vaccination, respectively. Seroboosting (two- to fourfold increase in EIA titers of individuals seropositive by the whole-RV EIA before vaccination) was usually accompanied by a shift in the IgG immunoblot pattern from a single (E1) to multiple (E1-E1, E1-C, or E1-E2-C) specificities, suggesting exposure of new epitopes as a result of viral replication.

Adolescent↗

PubMed1996

Changes in breath 13CO2/12CO2 during exercise of different intensities.

The measurement of breath 13CO2/12CO2 is commonly used during exercise to evaluate the oxidation rate of exogenous carbohydrates enriched in 13C. The aim of this study was to investigate whether exercise itself affects the 13C/12C ratio in expired air CO2 in relation to exercise intensity. The relative abundance of 13C and 12C in expired air CO2 was determined by isotoperatio mass spectrometry and expressed as delta 13C (in %o) by using Craig's formula and calibrated standards. Five healthy young men exercised on a treadmill after an overnight fast during > or = 105 min on four occasions and in a randomized order. Work rates were performed at approximately 30, 45, 60, and 75% of their maximal O2 uptake (VO2max). Delta 13C in expired air CO2 and respiratory exchange ratio (RER) were determined every 15 or 30 min during exercise. At 30 and 45% VO2max, a slight and not statistically significant increase in delta 13C was observed at 30 min. In contrast, at 60 75% VO2max, the rise was statistically significant and averaged 0.83 and 0.99%o, respectively. Average delta 13C (between 0 and 105 min) progressively increased with the intensity of exercise. Individual values of delta 13C and RER were positively correlated (r = 0.653, P = 0.002) as were values of delta 13C and endogenous carbohydrates utilized (r = 0.752, P < 0.001). Factitious or "pseudooxidation" of a 13C-enriched exogenous glucose load (indeed noningested) was calculated from the changes in expired air delta 13C. Over the whole period of exercise it was not statistically significant at 30 and 40% VO2max. However, over the first 60 min of exercise, such pseudooxidation of exogenous glucose was significant at 30 and 45% VO2max. In conclusion, by modifying the mix of endogenous substrates oxidized, exercise at 60% VO2max and above significantly increases the 13C/12C ratio in expired air CO2. At these intensities, this could lead to overestimation of the oxidation of 13C-labeled substrates given orally. At lower intensities of exercise, such overestimation is much smaller an affects mainly the values recorded during the initial part of the exercise bout.

Adult↗

PubMed1995

Structure of the catalytic region of human complement protease C1s: study by chemical cross-linking and three-dimensional homology modeling.

C1s is a multidomain serine protease that is responsible for the enzymatic activity of C1, the first component of the classical pathway of complement. Its catalytic region (gamma-B) comprises two contiguous complement control protein (CCP) modules, IV and V (about 60 residues each), a 15-residue intermediary segment, and the B chain (251 residues), which is the serine protease domain. With a view to identify domain-domain interactions within this region, the gamma-B fragment of C1s, obtained by limited proteolysis with plasmin, was chemically cross-linked with the water-soluble carbodiimide 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide; then cross-linked peptides were isolated after CNBr cleavage and thermolytic digestion. N-Terminal sequence and mass spectrometry analyses allowed us to identify two cross-links between Lys 405 of module V and Glu 672 of the B chain and between Glu 418 of the intermediary segment and Lys 608 of the B chain. Three-dimensional modeling of the CCP modules IV and V and of the catalytic B chain was also carried out on the basis of their respective homology with the 16th and 5th CCP modules of complement factor H and type I serine proteases. The information provided by both the chemical cross-linking studies and the homology modeling enabled us to construct a three-dimensional model for the assembly of the C-terminal part of the gamma-B region, comprising module V, the intermediary segment, and the B chain. This model shows that module V interacts with the serine protease B chain on the side opposite to both the activation site and the catalytic site. Functional implications of this interaction are discussed in terms of the possible role of module V in the specific recognition and positioning of C4, one of the two substrates of C1s.

Amino Acid Sequence↗

PubMed1995

Analysis of the N-linked oligosaccharides of human C1s using electrospray ionisation mass spectrometry.

Information on the structures of the oligosaccharides linked to Asn residues 159 and 391 of the human complement protease C1s was obtained using mass spectrometric and monosaccharide analyses. Asn159 is linked to a complex-type biantennary, bisialylated oligosaccharide NeuAc2 Gal2 GlcNAc4 Man3 (molecular mass = 2206 +/- 1). Asn391 is occupied by either a biantennary, bisialylated oligosaccharide, or a triantennary, trisialylated species NeuAc3 Gal3 GlcNAc5 Man3 (molecular mass = 2861 +/- 1), or a fucosylated triatennary, trisialylated species NeuAc3 Gal3 GlcNAc5 Man3 Fuc1 (molecular mass = 3007 +/- 1), in relative proportions of approximately 1:1:1. The carbohydrate heterogeneity at Asn391 gives rise to three major types of C1s molecules of molecular masses 79,318 +/- 8 (A), 79,971 +/- 8 (B), and 80,131 +/- 8 (C), with an average mass of 79,807 +/- 8. A minor modification, yielding an extra mass of 132 +/- 2, is also detected within positions 1-153.

Asparagine↗

PubMed1995

High-performance liquid chromatographic assays with fluorometric detection for mivacurium isomers and their metabolites in human plasma.

Two high-performance liquid chromatographic assays coupled with fluorometric detection have been developed for the determination of mivacurium isomers (trans-trans, cis-trans and cis-cis) and their monoester and alcohol metabolites in human plasma. A novel solid-phase extraction procedure allowed good recovery of the mivacurium isomers (mean 98%) and their monoester metabolites (mean 83%), whereas the alcohol metabolites were analyzed after direct precipitation of plasma proteins. For all analytes, these assays proved to be sensitive (LOQ 3.9-15.6 ng/ml), reproducible (C.V. < 15%) and accurate (> 94%) over the therapeutic range of concentrations of mivacurium and its metabolites. These two methods were applied successfully to a pharmacokinetic study of mivacurium after a bolus dose of 0.15 mg/kg in anesthetized patients.

Chromatography, High Pressure Liquid↗

PubMed1995

Keratinocytes modulate the biosynthetic phenotype of dermal fibroblasts at a pretranslational level in a human skin equivalent.

In this study we investigated the influence of keratinocytes on the phenotype of fibroblasts in an in vitro human skin equivalent. Keratinocytes were seeded at the surface of fibroblast-populated mechanically restrained type I collagen gels (lattices). Lattices without keratinocytes were handled in parallel as controls. After 2 and 4 days in culture, the keratinocyte layer was removed and the steady-state level of the mRNA for the main extracellular matrix macromolecules and interstitial collagenase produced by the fibroblasts was measured by Northern and dot blot analysis. A 50% decrease in the amount of procollagen type I and type III mRNAs was observed after 2 and 4 days of coculture while collagenase gene expression was upregulated by 300% when compared with control lattices. No significant modulation of type IV and type VI collagen, elastin or laminin B1 mRNA levels was found. Fibronectin mRNA levels in fibroblasts were significantly increased only on day 4. All the observed changes could be reproduced using a conditioned medium collected from a lattice covered with keratinocytes added to a lattice containing fibroblasts alone. These results indicate that in an in vitro reconstituted skin, keratinocytes are able to modulate the biosynthetic phenotype of fibroblasts at a pretranslational level through a paracrine signalling pathway.

Adult↗

PubMed1995

Antiestrogenic activity of two 11 beta-estradiol derivatives on MCF-7 breast cancer cells.

Two 11 beta-derivatives of estradiol (E2) were tested for their potential antiestrogenic activity in the MCF-7 breast cancer model: one contained a phenoxydimethylaminoethyl side-chain (RU 39,411), the other a pentafluoropentylsulfinyl side-chain (RU 58,668). The former compound displayed mixed estrogenic/antiestrogenic properties, while the latter indicated only an antiestrogenic activity. Both the compounds produced a growth inhibition of MCF-7 cells at doses related to their binding affinity for the estrogen receptor (ER); E2 suppressed this inhibition. The compounds also down-regulated the estrogen binding capacity of the cells but failed to reduce ER mRNA levels, indicating that the grafting of their side-chains prevented this antagonistic effect usually observed with steroidal estrogens. Assessment of ER levels by enzyme immunoassay revealed a marked increase with RU 39,411 and a decrease with RU 58,668; different mechanisms of action should, therefore, be considered. Finally, the estrogenic activity of RU 39,411 was demonstrated by its strong ability to induce synthesis of the progesterone receptor; RU 58,668 failed to display this agonistic activity.

Antineoplastic Agents↗

PubMed1995

Exogenous glucose oxidation during exercise in relation to the power output.

In order to study the influence of the power output on the oxidation rate of exogenous glucose and on the contribution of the various substrates to the energy demand, we combined the use of artificially enriched 13C-glucose with classical indirect calorimetry during uphill treadmill exercise. Six young male healthy subjects underwent three exercise bouts, in a randomized order and at least two weeks apart, at a low (45% VO2max, 1822 +/- 194 ml O2/min for 4 hours), moderate (60% VO2max, 2582 +/- 226 ml O2/min for 3 hours), and high intensity (75% VO2max, 3036 +/- 287 ml O2/min for 2 hours). After 10 min of exercise, each subject ingested 100 g of artificially 13C-labelled glucose dissolved in 400 ml of water. Over the four hours of the exercise at 45% VO2max, the amount of exogenous glucose oxidized was 89.5 +/- 5.9 g from the 100 g ingested. In all exercise bouts, the oxidation of exogenous glucose already began during the first 30 min after ingestion and peaked at 120 min. The maximum oxidation rates averaged 0.64 +/- 0.07, 0.75 +/- 0.04, and 0.63 +/- 0.08 g/min, and the mean amounts of exogenous glucose oxidized over the first two hours averaged 51.7 +/- 8.0, 61.5 +/- 6.6 and 50.9 +/- 8.45 g, at 45, 60 and 75% VO2max respectively. The contribution of the oxidation of exogenous glucose to the total energy supply progressively decreased when the power output increased, from 19.6 to 12.2%. In the meantime, the contribution of total carbohydrates (exogenous+endogenous) progressively increased from 55.1 to 77.8% while the contribution of lipids decreased from 35.5 to 16.6%. In conclusion, exogenous glucose ingested during exercise is largely oxidized and strongly contributes to the energy supply. The oxidation rate first increases with the power output, but levels off or even decreases at high intensity exercise.

Adult↗

PubMed1995

[Chronic urticaria and autoimmune thyroid diseases].

INTRODUCTION: Chronic urticaria results from multiple interactive causes. For certain authors dythyroidism is significantly more frequent in patients with chronic urticaria, often associated with other autoimmune disorders. PATIENTS AND METHODS: This study included 45 patients with chronic urticaria (29 men and 16 women, mean age 45.6 years). All underwent clinical examination for thyroid disease with assay of free T3 and T4, ultrasensitive TSH and antimicrosome and antithyroperoxidase antithyroid antibodies. RESULTS: Among the 45 patients, 8, all women, had an autoimmune thyroid disease: Graves' disease (n = 1), juvenile chronic thyroiditis (n = 1), autoimmune disease n = 6). All had goiter but most had no clinical sign suggestive of hypothyroidism or hyperthyroidism. Other clinical features or laboratory findings suggested autoimmune disorders in 3. DISCUSSION: These results suggest that a complete thyroid examination with hormone assay, especially in women, should be performed in patients with chronic urticaria. TSH and antiperoxidase antibodies appear to be the most sensitive and specific assays in patients with autoimmune thyroid disease.

Adult↗

PubMed1994

Mapping of linear antigenic sites on the S glycoprotein of a neurotropic murine coronavirus with synthetic peptides: a combination of nine prediction algorithms fails to identify relevant epitopes and peptide immunogenicity is drastically influenced by the nature of the protein carrier.

The elucidation of the antigenic structure of the S glycoprotein of murine coronaviruses will provide further understanding of the complex pathogenicity of these viruses. In order to identify linear antigenic determinants, the primary structure of the S glycoprotein of murine hepatitis virus strain A59 was analyzed with a combination of nine epitope prediction algorithms. Fifteen potential epitopes were synthesized chemically and injected into BALB/c mice to study their biological relevance. This approach failed to identify novel important epitopes. Furthermore, the algorithms were unable to identify as antigenic the previously mapped immunodominant epitope A [C. Daniel, R. Anderson, M. J. Buchmeier, J. O. Fleming, W. J. M. Spaan, H. Wege, and Talbot, P. J. (1993). J. Virol. 67, 1185-1194]. Interestingly, peptide A coupled to KLH induced an immune response that simulated the immune response induced by the corresponding region of the protein much more accurately than when the same peptide was coupled to BSA. This included drastically enhanced competition with monoclonal antibodies and protection from virus challenge. These findings emphasize the shortcomings of amino acid sequence-based epitope prediction algorithms and demonstrate the critical importance of the carrier when synthetic peptides are considered as potential vaccines.

Amino Acid Sequence↗

PubMed1994

Regulation of lymphocyte calcitonin receptors by interleukin-1 and interleukin-6.

We have reported the existence of specific and high-affinity calcitonin (CT) receptors on normal human T-lymphocytes. Because of the increasingly recognized importance of interleukin-1 (IL-1) and IL-6 in the control of bone metabolism, we have examined their influence on the binding parameters of labeled salmon calcitonin (sCT) on lymphocytes. After a 24-hour incubation, IL-1 at 100-5000 U/ml and IL-6, at 1-1000 U/ml, decreased the apparent number of CT binding sites (Bmax) on T-lymphocytes. The effects of IL-6 on purified T-lymphocytes were dose related and 100 U of IL-6/ml reduced sCT binding to 57 +/- 16% (mean +/- SD) of the control values (n = 6). There was no significant change in CT binding affinity (Kd, 0.71 +/- 0.54 x 10(-10) M for controls versus 0.90 +/- 0.55 x 10(-10) M after IL-6) and the decrease in Bmax was reversible after 48 hours. The effects of IL-1 appeared to be mediated through an increased production of IL-6 as they were neutralized by a polyclonal antiserum against IL-6. Added alone, the antiserum caused a slight increase in the apparent number of CT binding sites on T-lymphocytes to 115 +/- 5% of control values (n = 3). In summary, IL-1 and IL-6 can induce a marked apparent loss of CT binding sites on normal T-lymphocytes at concentrations known to be active on bone metabolism. The contributions of our observations to the osteolytic activity of these cytokines deserve further investigation.

Animals↗

PubMed1994

Availability of glucose ingested during muscle exercise performed under acipimox-induced lipolysis blockade.

This study investigated the percentage of carbohydrate utilization than can be accounted for by glucose ingested during exercise performed after the ingestion of the potent lipolysis inhibitor Acipimox. Six healthy male volunteers exercised for 3 h on a treadmill at about 45% of their maximal oxygen uptake, 75 min after having ingested 250 mg of Acipimox. After 15-min adaptation to exercise, they ingested either glucose dissolved in water, 50 g at time 0 min and 25 g at time 60 and 120 min (glucose, G) or sweetened water (control, C). Naturally labelled [13C]glucose was used to follow the conversion of the ingested glucose to expired-air CO2. Acipimox inhibited lipolysis in a similar manner in both experimental conditions. This was reflected by an almost complete suppression of the exercise-induced increase in plasma free fatty acid and glycerol and by an almost constant rate of lipid oxidation. Total carbohydrate oxidation evaluated by indirect calorimetry, was similar in both experimental conditions [C, 182, (SEM 21); G, 194 (SEM 16) g.3 h-1], as was lipid oxidation [C, 57 (SEM 6); G, 61 (SEM 3) g.3 h-1]. Exogenous glucose oxidation during exercise G, calculated by the changes in 13C:12C ratio of expired air CO2, averaged 66 (SEM 5) g.3 h-1 (19% of the total energy requirement). Consequently, endogenous carbohydrate utilization was significantly smaller after glucose than after placebo ingestion: 128 (SEM 18) versus 182 (SEM 21) g.3 h-1, respectively (P < 0.05). Symptoms of intense fatigue and leg cramps observed with intake of sweet placebo were absent with glucose ingestion.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗

PubMed1994

Analysis of overlapping T- and B-cell antigenic sites on rubella virus E1 envelope protein. Influence of HLA-DR4 polymorphism on T-cell clonal recognition.

A CTL antigenic site located between residues 273 and 291 of the E1 envelope protein of RV was identified by 51Cr-release assays employing SPs. Two E1-specific CTL clones were examined for immune recognition of RV wild-type and attenuated vaccine strains and recombinant E1 protein. The exact sequence (273-284) recognized by both clones was delineated by using truncated and overlapping SPs covering these residues. The defined T-cell site overlapped almost completely with a virus neutralizing antibody-binding site previously identified with mouse monoclonal and human antibodies. A series of single aa-substituted SP analogues of E1(273-284) was used to define residues critical for T-cell recognition. Using EBV-BL displaying different HLA-DR haplotypes and -DR4 subtypes as targets to determine MHC class II restriction elements, immune recognition by both T-cell clones was shown to be associated with HLA-DR4. Three HLA-DR4 subtypes (DR4Dw13A, DR4Dw13B, and DR4KT2) sharing a common residue, glutamic acid at position 74 in their beta 1 chains, were able to present SP E1(273-284) to the T-cell clones.

Amino Acid Sequence↗

PubMed1994

Use of synthetic peptides to map regions of rubella virus capsid protein recognized by human T lymphocytes.

Synthetic peptides (SPs), 18-29 amino acids long, representing selected sequences of rubella virus (RV) capsid (C) protein were used in lymphocyte proliferation assays to identify antigenic regions recognized by T lymphocytes from healthy RV-reactive adults. Four SPs, C(1-29), C(90-114), C(108-134) and C(255-300), stimulated proliferation of peripheral blood mononuclear cells and RV-specific T-cell lines from the same donors. C(1-29V), an SP analogue containing an RA27/3 RV vaccine strain sequence, stimulated higher levels of proliferation in T cells obtained from RV-vaccinated subjects than did the comparable wild-type (M33 strain) RV sequence.

Adult↗

PubMed1994

Comparison of novel synthetic peptide-based DETECT-RUBELLA enzyme immunoassays with Enzygnost and IMx for detection of rubella-specific immunoglobulin G.

Enzyme immunoassays (EIAs) using synthetic peptides SP-E1 and SP-E1E2 (DETECT-RUBELLA [Bio-Chem]) were compared with two viral lysate-based EIAs (Enzygnost [Behring] and IMx [Abbott]) for the detection of rubella virus-specific immunoglobulin G antibodies. Sensitivities of 94.7, 100, 98.6, and 100% and specificities of 100, 97.4, 100, and 73.7% were found for the SP-E1, SP-E1E2, Enzygnost, and IMx EIAs, respectively.

Antibodies, Viral↗