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P Lucas

Publications and source records attributed to P Lucas.

At least 37 records · Page 2Linked to original sources
PubMed2000

Membrane raft microdomains mediate lateral assemblies required for HIV-1 infection.

HIV-1 infection triggers lateral membrane diffusion following interaction of the viral envelope with cell surface receptors. We show that these membrane changes are necessary for infection, as initial gp120-CD4 engagement leads to redistribution and clustering of membrane microdomains, enabling subsequent interaction of this complex with HIV-1 co-receptors. Disruption of cell membrane rafts by cholesterol depletion before viral exposure inhibits entry by both X4 and R5 strains of HIV-1, although viral replication in infected cells is unaffected by this treatment. This inhibitory effect is fully reversed by cholesterol replenishment of the cell membrane. These results indicate a general mechanism for HIV-1 envelope glycoprotein-mediated fusion by reorganization of membrane microdomains in the target cell, and offer new strategies for preventing HIV-1 infection.

Animals↗
PubMed1999

Influence of alveolar ventilation changes on calculated gastric intramucosal pH and gastric-arterial PCO2 difference.

OBJECTIVE: To evaluate the influence of changes in alveolar ventilation on the following tonometry-derived variables: gastric intramucosal CO2 tension (PtCO2), gastric arterial CO2 tension difference (PgapCO2), gastric intramucosal pH (pHi) and arterial pH-pHi difference (pHgap). DESIGN: Clinical prospective study. SETTING: A medical intensive care unit in a university hospital. PATIENTS: Ten critically ill, mechanically ventilated patients requiring hemodynamic monitoring with pulmonary artery catheter. INTERVENTIONS: Gastric tonometer placement. A progressive increase in tidal volume (V(T)) from 7 to 10 ml/ kg followed by an abrupt return to baseline V(T) level. MEASUREMENTS AND MAIN RESULTS: Tonometer saline PtCO2 and hemodynamic data were collected hourly at various V(T) levels: H0 and H0' (baseline V(T) = 7 ml/kg), H1 (V(T) = 8 ml/kg), H2 (V(T) = 9 ml/kg), H3 (V(T) = 10 ml/kg), H4 (baseline V(T)). During the "hyperventilation phase" (H0-H3), pHi (p<0.01) and pHgap (p<0.05) increased but PgapCO2 remained unchanged. Cardiac output (CO) was not affected by ventilatory change. During the "hypoventilation phase" (H3-H4), pHi fell from 7.27+/-0.11 to 7.23+/-0.09 (p<0.01) and PgapCO2 decreased from 16+/-5 mm Hg to 13+/-4 mm Hg (p<0.05). V(T) reduction was associated with a significant cardiac output elevation (p<0.05). CONCLUSIONS: PaCO2 and PtCO2 are similarly influenced by the changes in alveolar ventilation. Unlike pHi, the PgapCO2 is not affected by ventilation variations unless CO changes are associated.

Blood Gas Analysis↗
PubMed1999

Antibody repertoire against HIV-1 gp120 triggered in nude and normal mice by GM-CSF/gp120 immunization.

Granulocyte-macrophage colony stimulating factor (GM-CSF) facilitates the induction of primary immune responses by activating and recruiting antigen-presenting cells (APC), which efficiently present antigen determinants to Th cells. We have derived a functional GM-CSF/gp120 chimeric protein that, following immunization in soluble, adjuvant-independent form in normal mice, triggers highly specific, high affinity anti-gp120 antibodies. In contrast, nude mice respond with mutated, polyreactive, low affinity antibodies that mature further and increase in affinity in T cell-reconstituted nude mice. Anti-gp120 antibody production in nude mice is mediated principally by GM-CSF/gp120-triggered IL-4 production, since neutralizing anti-IL-4 abrogates the in vivo response. The anti-gp120 antibody response in normal, nude and T cell-reconstituted nude mice is encoded at a remarkably high frequency by the VH81X and VH7183 genes, a family used notably during fetal life and, when expressed at the adult stage, associated with autoimmune disease. We conclude that HIV gp120 binds and selects a subpopulation of developing B cells expressing a set of VH genes associated with immunodeficiency and autoimmunity.

Animals↗
PubMed1999

Development of multiple calcium channel types in cultured mouse hippocampal neurons.

The development of multiple calcium channel activities was studied in mouse hippocampal neurons in culture, using the patch-clamp technique. A depolarizing pulse (40-50 ms duration) from the holding potential of -80 mV to levels more depolarized than -40 mV produced a low threshold T-type current. The T-type current was observed in 52% of four days in vitro neurons. The number of neurons which expressed T-type current decreased with age of culture, so that the current was detected in only 18% of neurons after 16 days in vitro. The T-type current densities varied between 1.9 pA/pF and 3.29 pA/pF in the mean values during the period studied (4-16 days in vitro). A depolarizing pulse from -80 mV to levels more depolarized than -35 mV evoked a high threshold calcium channel current. The high threshold current density increased in the mean values from 3.9 pA/pF in four days in vitro neurons to 28 pA/pF in 16 days in vitro neurons. We have then examined the effect of nifedipine, omega-Agatoxin IVA and omega-conotoxin GVIA on the high threshold current. Nifedipine (1-5 microM) sensitive current density stayed in the range of 1.9-2.1 pA/pF during 4-16 days in vitro, while omega-Agatoxin IVA (200 nM) sensitive current density increased in the mean values from 1.54 pA/pF in four days in vitro neurons to 21.5 pA/pF in 16 days in vitro neurons. The omega-conotoxin GVIA sensitive N-type channel current was maximum at eight days in vitro (5.44 pA/pF) and it reduced progressively to reach almost half (2.46 pA/pF) in 16 days in vitro neurons. These results showed that diverse subtypes of calcium channels change in density during the early period of culture. We suggest that the temporal expression of each type of channel may be linked to the development of neural activities.

Amiloride↗
PubMed1999

Natural human antibodies retrieved by phage display libraries from healthy donors: polyreactivity and recognition of human immunodeficiency virus type 1gp120 epitopes.

We have characterized the human natural antibody repertoire that contains antibodies recognizing the human immunodeficiency virus type 1 (HIV-1) gp120. A panel of monovalent antigen-binding fragments (Fab) selected from IgM and IgG isotype libraries generated from peripheral blood mononuclear cells (PBMC) of a healthy, HIV-1 noninfected individual was analysed, reflecting that only IgM, but not IgG, Fab were able to recognize HIV-1 gp120. The IgM Fab antibodies were not restricted to any particular heavy chain variable region (VH) germ line gene. However, the recognition of gp120 is associated to polyreactive antibodies and all display low-affinity interaction. This correlates with the absence of any maturation process as somatic mutation or isotype switch as the nucleotide sequence analysis of the variable regions reveals they are expressed near to germline configuration. In addition, none of the antibodies showed any neutralizing activity on HIV-1-infected lymphocytes, reflecting that the natural anti-gp120 repertoire is not sufficient to neutralize HIV infection.

Antibodies, Monoclonal↗
PubMed1999

A human immunodeficiency virus type 1 Env-granulocyte-macrophage colony-stimulating factor fusion protein enhances the cellular immune response to Env in a vaccinia virus-based vaccine.

Vaccinia virus (VV) infection induces protective T- and B-cell responses, making recombinants based on VV good candidates for the development of effective vaccines to other viruses. VV recombinants expressing the human immunodeficiency virus (HIV) envelope protein (Env) have been generated in several laboratories and shown to induce anti-HIV cellular and humoral immune responses in vaccinated humans and in chimpanzees. To increase the immunogenicity of the Env antigen, a VV recombinant was generated that expresses a chimeric antigen consisting of the Env protein fused to an immunostimulatory cytokine, granulocyte-macrophage colony-stimulating factor (GM-CSF). The chimeric protein retained GM-CSF biological activity when expressed by this recombinant virus (VV-GM-gp120) in cells infected in vitro. Infection of BALB/c mice with VV-GM-gp120 triggered a higher HIV-specific cellular immune response, as measured by interferon-gamma production, than that induced by a VV recombinant expressing the native Env protein. Moreover, although anti-gp120 antibody titres were similar in sera from mice inoculated with either of the VV recombinants, immunization with the recombinant expressing the fusion protein elicited antibodies against a broader spectrum of Env epitopes. These results indicate that HIV Env antigen fusion to GM-CSF provides a means to improve the anti-HIV immune response.

AIDS Vaccines↗
PubMed1999

Pharmaceutical and biological properties of poly(amino acid)/DNA polyplexes.

Physicochemical properties of polyplexes formed between pRSVlacZ and poly(amino acid)s were investigated as a paradigm of more complex, synthetic virus-like, DNA delivery systems, that are of interest to many gene delivery laboratories. We observed the interaction between polymer and DNA using ethidium exclusion, and determined the size distributions and the zeta potentials of polyplexes. We correlated these properties with their fundamental interactions with cultured B16 murine melanoma cells, and the resulting efficiency of transfection. A variety of poly(amino acid)s each condensed DNA to produce particles with mean hydrodynamic diameters of approximately 100 nm (a typical span of a population was 80-120nm). Poly(amino acid) polyplexes were unstable in electrolyte solutions such as cell culture media. The apparent particle size increased in electrolyte, depending on the charge ratio, to diameters up to 700 nm. This was thought to be due to aggregation, since neutral particles were most sensitive. When the charge ratio (+/-) exceeded unity polyplexes had positive zeta potentials (which peaked at approximately +30 mV), bound non-specifically to cells, were internalised and in the presence of an endosomolytic agent were able to transfect cells. Though all cationic poly(amino acid)s investigated formed polyplexes with similar physical properties, their biological properties were significantly different. Polyplexes prepared with poly-L-ornithine were the most effective transfection agents, but poly(lys-co-ala, 1: 1) systems appeared to be inactive. This may reflect the differences in uncoupling of DNA and polymer, which is expected to be necessary for passage through the nuclear pore. Uncoupling of polycation and DNA was investigated by exposing the complexes to dextran sulphate. Release of DNA was detected by increased fluorescence at 600 nm in the presence of ethidium. Release of DNA was incomplete from polyplexes formed with high molecular weight polylysine. This may explain the lower levels of transfection observed with high molecular weight polylysine. The significance of these observations for design of advanced non-viral gene delivery systems is discussed.

Amino Acids↗
PubMed1999

[Driver's licence: responsibility, ethics and deontology].

The new law enacted on march 23rd 1998 regarding driving-licence, which became effective on october 1st 1998, modifies the method for obtaining the different types of driving-licences. Its 6th annex enumerates a list of physical impairments and pathologies implying the denial, the limitation or the withdrawal of the driving-licence; it determines if a medical advice, and if the case which kind of medical advice, is required. This leads to new situations, for which no jurisprudence has been established to date, where not only the responsibility of the licence's applicant but also that of the examining physician could be suspected. The physicians are also confronted to ethical an deontological problems on which the medical authorities have not always given an opinion. This paper examines them in order to give some bases for reflexion and discussion.

Automobile Driver Examination↗
PubMed1999

[Endemic dracunculiasis in new villages: re-emergence of the disease or failure of the surveillance system].

According to data sent to Bamako in March 1998 for the annual review of National Programs for the Eradication of the Guinea Worm (NPEGV), dracunculiasis was observed in 211 villages in Burkina Faso in 1997. Of this total, 110 (52 p. 100) were new villages not previously reporting dracunculiasis. A study focusing on these new villages was carried out in June 1998. The aim of the study was to evaluate the quality of the disease monitoring system and determine if endemicity was spreading to new villages. A twofold method was used first to analyze the data used in the geographic information section of the disease monitoring system and second to test the validity of this data in field surveys. Findings showed the actual number of new villages showing endemicity could be reduced from 46 p. 100 to 12 p. 100 since 67 p. 100 of the new villages had in fact previously reported endemicity but had been wrongly eliminated from monitoring records. This probably long-standing problem has been uncovered with a decrease in the number of villages showing endemicity and an increase in prevention activities. In addition this study identified several confounding factors and defects in disease monitoring and data reporting techniques. Based on this study, several recommendations were made to improve the quality of the disease monitoring system.

Bias↗
PubMed1999

A diagnostic advice system based on pathophysiological models of diseases.

Medical decision-support systems in which uncertainty plays an essential role are increasingly based on the formalism of probabilistic networks. Although this formalism is very powerful, the construction of actual networks is not straightforward, and requires the availability of clearly structured medical domain models as a starting point. In this paper it is argued that medical pathophysiological knowledge constitutes a good start for the development of such models, even though pathophysiological knowledge is semantically different from probabilistic knowledge. Two models concerning anaemia, which are part of a broad system covering the domain of anaemia, are discussed to illustrate the general approach.

Anemia, Pernicious↗
PubMed1999

A probabilistic approach to improved antibiotic therapy.

PTA is a decision-theoretic expert system that aims to assist clinicians in diagnosing and treating patients with pneumonia in the intensive care unit. Its underlying probabilistic network model includes knowledge for diagnosing pneumonia on the basis of the likelihood of tracheobronchial-tree colonisation by pathogens, and symptoms and signs actually present in the patient. Optimal antibiotic therapy is selected by balancing the expected efficacy of treatment, which is related to the likelihood of pathogen-specific pneumonias, against costs and side effects of treatment. In this article, the model underlying the system and results of a preliminary evaluation are described.

Anti-Bacterial Agents↗
PubMed1998

Functional characterization of a new class of odorant-binding proteins in the moth Mamestra brassicae.

A new protocol of binding assay allowed us to functionally characterize two additional odorant-binding proteins in antennae of the moth Mamestra brassicae. These proteins have no N-terminal sequence homology with the moth pheromone-binding proteins and general odorant-binding proteins previously described. One of the two proteins designated MbraAOBP2 is between 60 and 73% similar in N-terminal to several proteins characterized in chemosensory organs of Diptera, Hymenoptera, Lepidoptera, and Phasmids, indicating that these proteins constitute a new group of odorant-binding proteins. A particularly high similarity between MbraAOBP2 and ejaculatory bulb specific protein III of Drosophila suggested that vaccenyl acetate could be a specific ligand for these proteins. As a matter of fact, MbraAOBP2 bound vaccenyl acetate in vitro, but we failed to detect any receptor cell in long and short sensilla trichodea of males. This protocol could be used as a rapid method to identify new odorant-binding proteins in chemosensory organs or tissues.

Amino Acid Sequence↗
PubMed1998

Suppression of HIV-1 infection in linomide-treated SCID-hu-PBL mice.

BACKGROUND: Proinflammatory cytokine overproduction, as well as synthesis of the inducible form of nitric oxide synthase (iNOS), are known to play a major role in HIV-1-triggered disease. AIDS patients show increased serum tumour necrosis factor (TNF)-alpha and interferon (IFN)-gamma levels, which synergize with HIV-1-produced nitric oxide (NO) to augment viral replication. Linomide has strong immunomodulatory effects in animals and humans, yielding promising clinical benefits in several pathological disorders including septic shock and autoimmune disease, processes largely mediated by overproduction of these cytokines. In peripheral T cells, linomide also prevents apoptosis triggered by a variety of stimuli, including superantigens, dexamethasone and vaccinia virus. DESIGN AND METHODS: Linomide inhibits production of proinflammatory cytokines such as TNF-alpha, interleukin-1 beta and IFN-gamma, as well as iNOS synthesis. The SCID-hu-PBL mouse model was used to analyse the effect of linomide on HIV-1 infection. T-cell frequency was characterized in reconstituted animals, and the frequency of infected mice and viral load of infected animals were studied. RESULTS: Linomide promotes an increase in human CD4+ T-cell counts in the peritoneal cavity of HIV-1-infected, linomide-treated mice. Linomide also prevents human TNF-alpha and IFN-gamma production, as well as iNOS expression and affects the viral load, promoting potent suppression of HIV-1 infectivity as detected in peritoneal cavity and spleen. CONCLUSIONS: The combination of linomide's properties, namely, blockage of proinflammatory cytokine and NO production, as well as prevention of apoptosis, is of paramount interest, making linomide a potential candidate for combating HIV-1 infection or preventing some of its associated pathological manifestations.

Adjuvants, Immunologic↗
PubMed1998

Polycation-DNA complexes for gene delivery: a comparison of the biopharmaceutical properties of cationic polypeptides and cationic lipids.

DNA plasmids formed particulate complexes with a variety of cationic polyamino acids and cationic lipids, which were used to transfect mammalian cells in culture. Complexation was studied by assaying for exclusion of ethidium using a fluorometric assay, which indicated that complexation with cationic polyamino acids took place with utilisation of the majority of charged functional groups. The particle sizes and zeta potentials of a range of complexes were determined. Generally polyamino acids formed uniform particles 80-120 nm in diameter in water, but their particle size increased on dilution of the particles in electrolytes or cell culture media. The efficiency of transfection was compared using complexes of pRSVlacZ, a reporter construct which expressed beta-galactosidase under the control of the Rous sarcoma virus promoter. Positively charged DNA/polyamino acid complexes were taken up by cells but required an endosomolytic agent, such as chloroquine, to facilitate transfection. Polyornithine complexes resulted in the highest levels of expression, in comparison with other homopolyamino acids (polyornithine>poly-L-lysine=poly-D-lysine>polyarginine). Copolyamino acids of lysine and alanine condensed DNA but were less active in transfection experiments. Copoly(L-Lys, L-Ala 1:1) was inactive even in the presence of chloroquine. In contrast DNA/cationic lipid complexes transfected cells spontaneously, and chloroquine did not improve the extent of expression, rather it usually reduced efficiency. There was little correlation between comparative efficiencies of lipid complexes between cell lines suggesting that the nature of the cell membrane and differences in mechanisms of internalisation were determinants of efficiency. In an effort to explore better cell culture models for gene delivery, monolayers of Caco-2 cells were transfected in filter culture. As the cells differentiated and formed a polarized monolayer, expression of beta-galactosidase was reduced until at day 27 expression was not significantly different from basal activity. The Caco-2 filter culture model merits further attention as a model of gene delivery to epithelial surfaces, such as would be encountered in the lung after inhalation.

Animals↗
PubMed1998

Emergency removal of football equipment: a cadaveric cervical spine injury model.

STUDY OBJECTIVE: To determine the influence of football helmet and shoulder pads, alone or in combination, on alignment of the unstable cervical spine. METHODS: The alignment of the intact cervical spine in 8 cadavers was assessed radiographically under 4 different football equipment conditions: (1) no equipment, (2) helmet only, (3) helmet and shoulder pads, and (4) shoulder pads only. Each specimen was then surgically destabilized at C5-C6 to simulate a flexion-distraction injury. Repeat radiographs were obtained under the same 4 equipment conditions, and alignment of the unstable segment was analyzed. RESULTS: Before the destabilization, neutral alignment was maintained when both helmet and shoulder pads were in place. The "helmet only" condition caused a significant decrease in lordosis (mean, 9.6 +/- 4.7 degrees), whereas the "shoulder pads only" condition caused increased lordosis (13.6 +/- 6.3 degrees). After destabilization, the "helmet-only" condition demonstrated significant mean increases in C5-C6 forward angulation (16.5 +/- 8.6 degrees), posterior disc space height (3.8 +/- 2.3 mm), and dorsal element distraction (8.3 +/- 5.4 mm). CONCLUSION: Our flexion-distraction model demonstrated that immobilization of the neck-injured football player with only the helmet in place violates the principle of splinting the cervical spine in neutral alignment. By extrapolation to an extension-type injury, immobilization with only the shoulder pads left in place similarly violates this principle. In order to maintain a neutral position and minimize secondary injury to the cervical neural elements, the helmet and shoulder pads should be either both left on or both removed in the emergency setting.

Aged↗
PubMed1998

HIV-1 envelope protein gp120 triggers a Th2 response in mice that shifts to Th1 in the presence of human growth hormone.

Immunization of mice with HIV-1-gp120 results in predominant activation of the Th2 lymphocyte subset, leading to enhanced IL-4 production. Administration of human growth hormone at the time of gp120 immunization provokes a change in the cytokine production pattern, with lower IL-4 and higher gamma-IFN and IL-2 synthesis levels, indicating a preferential switch in stimulation from Th2 to Th1 cells. A growth hormone would thus be of great use for pharmacological intervention in those cases in which an infectious microorganism evades immune defenses by provoking a Th2 response. In addition, the ability of growth hormone to induce a Th1-type response upon vaccination with an HIV-antigen should be examined in the development of new therapeutic strategies or in the design of novel vaccines against HIV infection.

Animals↗