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Biomedical subjects

M Girard

Publications and source records attributed to M Girard.

At least 109 records · Page 6Linked to original sources
PubMed1995

Picornavirus internal ribosome entry segments: comparison of translation efficiency and the requirements for optimal internal initiation of translation in vitro.

On the basis of primary sequence comparisons and secondary structure predictions, picornavirus internal ribosome entry segments (IRESes) have been divided into three groups (entero- and rhinoviruses; cardio- and and aphthoviruses; and hepatitis A virus). Here, we describe a detailed comparison of the ability of IRESes from each group to direct internal initiation of translation in vitro using a single dicistronic mRNA (the only variable being the IRES inserted into the dicistronic region). We studied the influence of various parameters on the capacity of six different picornaviral IRESes, and the non-picornaviral hepatitis C virus IRES, to direct internal initiation of translation: salt concentration, the addition of HeLa cell proteins to rabbit reticulocyte lysate translation reactions, the presence of foot-and-mouth disease virus Lb or human rhinovirus 2A proteinase. On the basis of the characteristics of IRES-driven translation in vitro, the picornaviral IRESes can be classified in a similar manner to when sequence homologies are considered. IRESes from each of the three groups responded differently to all of the parameters tested, indicating that while all of these elements can direct internal ribosome entry, the functional requirements for efficient IRES activity vary dramatically. In the individual optimal conditions for translation initiation, the best IRESes were those from the cardio- and aphthoviruses, followed by those from the enteroviruses, which exhibited up to 70% of the efficiency of the EMCV element in directing internal initiation of translation.

Base Sequence↗
PubMed1995

[Vaccination against the human immunodeficiency virus].

Much progress has been made in recent years in the development of anti-VIH vaccines. Nearly 20 such vaccines have reached phase 1 clinical study in seronegative volunteers. The responses invoked by these vaccines are, however, mediocre, both in quality (lack of cross over neutralisation in isolated "wild" viruses) and in their levels and length of action. New formulations of vaccines are under study, but their development is long and difficult, and researchers are still disarmed by the problem of the variability of the virus. Several years of study, both clinical and fundamental, will be necessary before an effective vaccine against HIV-1 is available.

AIDS Vaccines↗
PubMed1995

Purification and renaturation of Japanese encephalitis virus nonstructural glycoprotein NS1 overproduced by insect cells.

The nonstructural protein NS1 of Japanese encephalitis virus is a major immunogen produced during flavivirus infection. However, the function of this protein has not been identified. To analyze its biochemical properties and evaluate its potential activity in the virus life cycle, the protein was produced in Spodoptera frugiperda insect cells (Sf9), using a recombinant baculovirus, and purified. As described previously by M. Flamand, V. Deubel, and M. Girard (1992, Virology 191, 826-836), a small fraction of the synthesized recombinant protein could mature into a dimer, whereas the major part was retained in intracellular aggregates. This insolubility was used to recover the protein in a purified form using a two-step procedure. Isolated inclusion bodies, in which NS1 constituted over 60% of the protein, were solubilized in 8 M urea. NS1 was further purified by reverse-phase HPLC and recovered at over 90% purity with an overall yield of over 60%. Conditions promoting reoxidation-renaturation of the purified protein were then investigated at a concentration of 100 micrograms/ml at pH 8. The presence of 8 M urea during reoxidation of NS1 with oxidized glutathione was essential prior to renaturation by dialysis to avoid reaggregation, the main side pathway of refolding in vitro. Three major species, all monomeric, were resolved by nonreducing SDS-PAGE. The form showing the lowest apparent molecular weight comigrated with native unreduced NS1 and was recognized by a monoclonal antibody directed against a conformational epitope strictly dependent on the native structure of the protein.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Dose-response relationships for edrophonium antagonism of mivacurium-induced neuromuscular block during N2O-enflurane-alfentanil anaesthesia.

The purpose of this study was to determine the dose-response relationships for edrophonium antagonism of mivacurium-induced neuromuscular block. Seventy-five ASA I or II adults were given mivarcurium 0.15 mg x kg(-1) followed by an infusion (7 micrograms x kg(-1) x min(-1) during alfentanil-propofol-N2O-enflurane anaesthesia. Train-of-four stimulation (TOF) was applied to the ulnar nerve every 20 sec and the response of the adductor policis was recorded (Relaxograph NMT-100, Datex, Helsinki, Finland). Mivacurium infusion was modified at five-minute intervals in order to keep the height of the first twitch in TOF(T1) at 5% of its control value. At the end of surgery, edrophonium (0.0, 0.125, 0.25, 0.5, or 1.0 mg x kg(-1) combined glycopyrrolate (0.0, 0.0012, 0.0025, 0.005, or 0.01 mg x kg(-1) were administered by random allocation. Edrophonium doses of 0.25, 0.5 and 1.0 mg x kg(-1) were different from placebo with regard to time to attain a TOF ratio (fourth twitch in TOF/T1) = 0.7 (13.8 +/- 4.5, 11.1 +/- 3.5, 11.4 +/- 3.0 vs 19.7 +/- 4.7 min P< 0.05). Doses of 0.5 and 1.0 mg x kg(-1) permitted faster recovery time of T1 from 10 to 95% (T10-95) than did placebo (7.5 +/- 3.8, 8.9 +/- 3.5 vs 14.5 +/- 5.0 min P<0.05). Edrophonium 0.5 mg x kg(-1) was different from placebo with regard to recovery time of T1 from 25 to 75% (T25-75) (3.3 +/- 2.0 vs 6.7 +/- 2.0 min P<0.05). Only edrophonium 0.5 mg x kg(-1) provided faster recovery than placebo with regard to all three indices. It is concluded that edrophonium 0.5 + glycopyrrolate 0.005 mg x kg(-1) allow the fastest recovery from a mivacurium-induced block during enflurane-N2O anaesthesia.

Adolescent↗
PubMed1995

Present status of vaccination against HIV-1 infection.

The development of an effective vaccine against the acquired immunodeficiency syndrome (AIDS) is a formidable task. The absence of any documented case of natural recovery from the disease raises the question of whether any vaccine against human immunodeficiency virus (HIV) could possibly be effective in preventing natural transmission of the virus. It also raises the issue of which type of immune response a vaccine should elicit to be protective. Many obstacles have to be overcome, including the considerable antigenic variability of the virus, its intracellular mode of transmission, its mucosal port of entry, and the persistent nature of the infection. The only animal model available for the development of prototype HIV-1 vaccines is the chimpanzee model. Chimpanzees infected with HIV-1 do not develop AIDS but remain persistently infected. This model, therefore, allows one to test for protection from infection, but not for protection from disease.

AIDS Vaccines↗
PubMed1995

Heterologous HIV-2 challenge of rhesus monkeys immunized with recombinant vaccinia viruses and purified recombinant HIV-2 proteins.

In an attempt to analyse the role of anti-envelope immunity in the protection of rhesus monkeys against an HIV-2 intravenous challenge, rhesus macaques were immunized twice with recombinant HIV-2 ROD vaccinia viruses (10(8) p.f.u. each) at days 0 and 30, followed by booster injections of purified HIV-2 proteins at months 8, 9, 15 and 27. One group of five macaques was immunized with the Gag, Pol, Vif and Nef antigens, whereas a second group received the same antigens with the addition of HIV-2 Env protein. Eight months after the last boost, the animals were challenged by intravenous injection of 100 AID50 of a monkey PBMC-grown stock of HIV-2 SBL. None of the animals was protected in spite of high humoral immune responses on day of challenge as determined by ELISA and Western Blot assays.

AIDS Vaccines↗
PubMed1995

Continuous intravenous administration of ketorolac reduces pain and morphine consumption after total hip or knee arthroplasty.

The purpose of this study was to determine the analgesic efficacy, opioid-sparing effect, and tolerability of ketorolac administered as an intravenous (i.v.) bolus followed by a continuous infusion after total hip or knee arthroplasty. After general anesthesia, patients received either placebo or ketorolac 30 mg i.v. as a bolus over 15-30 s followed by a continuous i.v. infusion of ketorolac 5 mg/h for 24 h. All patients received patient-controlled i.v. morphine with no background infusion. Patients were assessed at 2, 4, 6, and 24 postoperatively with respect to analgesia, morphine consumption, side effects, and blood loss. Patients receiving ketorolac reported were less sedated and required fewer antiemetics. There was no difference in blood loss. Patients receiving ketorolac reported better analgesia and used less morphine (35% for hips and 44% for knees) than those receiving placebo.

Aged↗
PubMed1995

Vaccine-induced protection of chimpanzees against infection by a heterologous human immunodeficiency virus type 1.

The extraordinary genetic diversity of human immunodeficiency virus type 1 (HIV-1) is a major problem to overcome in the development of an effective vaccine. In the most reliable animal model of HIV-1 infection, chimpanzees were immunized with various combinations of HIV-1 antigens, which were derived primarily from the surface glycoprotein, gp160, of HIV-1 strains LAI and MN. The immunogens also included a live recombinant canarypox virus expressing a gp160-MN protein. In one experiment, two chimpanzees were immunized multiple times; one animal received antigens derived only from HIV-1LAI, and the second animal received antigens from both HIV-1LAI and HIV-1MN. In another experiment, four chimpanzees were immunized in parallel a total of five times over 18 months; two animals received purified gp160 and V3-MN peptides, whereas the other two animals received the recombinant canarypox virus and gp160. At 3 months after the final booster, all immunized and naive control chimpanzees were challenged by intravenous inoculation of HIV-1SF2; therefore, the study represented an intrasubtype B heterologous virus challenge. Virologic and serologic follow-up showed that the controls and the two chimpanzees immunized with the live recombinant canarypox virus became infected, whereas the other animals that were immunized with gp160 and V3-MN peptides were protected from infection. Evaluation of both cellular and humoral HIV-specific immune responses at the time of infectious HIV-1 challenge identified the following as possible correlates of protection: antibody titers to the V3 loop of MN and neutralizing antibody titers to HIV-1MN or HIV-1LAI, but not to HIV-1SF2. The results of this study indicate that vaccine-mediated protection against intravenous infection with heterologous HIV-1 strains of the same subtype is possible with some immunogens.

AIDS Vaccines↗
PubMed1995

Attenuated Mengo virus: a new vector for live recombinant vaccines.

Several features make Mengo virus an excellent candidate for use as a vaccine vector. The virus has a wide host range, including rodents, pigs, monkeys, and most likely humans, and expresses its genome exclusively in the cytoplasm of the infected cell. Stable attenuated strains exist which are deleted for part of the 5' noncoding region of the genome. Here we report an attenuated Mengo virus recombinant, vLCMG4, that encodes an immunodominant cytotoxic T-lymphocyte epitope of the lymphocytic choriomeningitis virus (LCMV) nucleo-protein. vLCMG4 induced protective immunity against lethal LCMV infection after a single, low-dose immunization in BALB/c mice and elicited an LCMV-specific CD8+ cytotoxic T lymphocyte response. This demonstrates the potential of recombinant Mengo virus vaccines to confer protection against infectious diseases by the induction of cellular immune responses.

Amino Acid Sequence↗
PubMed1994

Attenuated Mengo virus as a vector for immunogenic human immunodeficiency virus type 1 glycoprotein 120.

Introduction of a sequence encoding 147 amino acids from human immunodeficiency virus type I (HIV-1) strain MN glycoprotein gp120 into the RNA genome of the stably attenuated Mengo virus strain vM16 yielded an infectious recombinant virus, vMLN450, which expressed the heterologous HIV-1 sequence along with the normal Mengo virus proteins. The HIV-1 gp120 sequence, fused to the amino terminus of the short, nonstructural Mengo virus leader polypeptide was recognized by a gp120 V3 loop-specific monoclonal antibody. When inoculated into mice, recombinant virus vMLN450 elicited a high-titer anti-HIV-1 antibody response as well as an HIV-1MN-specific cytotoxic cellular immune response. An anti-HIV-1 antibody response could also be detected in cynomolgus monkeys after a single immunization. We propose that attenuated Mengo virus can serve as an effective expression vector in cell systems and various animal species and offers another approach to the development of new, live recombinant vaccines.

Amino Acid Sequence↗
PubMed1994

Programmed cell death and AIDS: significance of T-cell apoptosis in pathogenic and nonpathogenic primate lentiviral infections.

We have proposed that inappropriate induction of programmed cell death (PCD) or apoptosis, a physiological cell-suicide process, may play a role in the pathogenesis of AIDS. This model has been supported by several reports of abnormal levels of PCD in vitro in both CD4+ and CD8+ T cells from human immunodeficiency virus type 1 (HIV-1)-infected persons. To further assess the significance of such a process in AIDS pathogenesis, in vitro PCD was compared in HIV-1-infected persons and in various primate models that allow discrimination between pathogenic chronic lentiviral infection either in the same species, such as rhesus macaques infected with different simian immunodeficiency viruses (SIV), or in different species, such as SIV-infected African green monkeys and HIV-1-infected chimpanzees. Abnormal levels of PCD in CD4(+)-T-cell-depleted peripheral blood mononuclear cells (PBMC), containing the CD8+ T cells, were observed in both pathogenic and nonpathogenic models. However, abnormal levels of PCD in the CD8(+)-T-cell-depleted PBMC, containing the CD4+ T cells, was only observed in the two models leading to AIDS: HIV-1-infected persons and rhesus macaques infected with a pathogenic strain of SIV. This suggests that inappropriate T-cell PCD in HIV-1-infected persons involves two distinct processes: one, concerning CD4+ T cells, is closely related to AIDS pathogenesis; and the other, concerning CD8+ T cells, may be a consequence of immune stimulation with no direct link to AIDS pathogenesis.

Acquired Immunodeficiency Syndrome↗
PubMed1994

[The use of 2-chloroprocaine for a combined lumbar plexus and sciatic nerve block].

A combination of the lumbar plexus and sciatic nerve blocks using 2-chloroprocaine (2-CP) for unilateral lower extremity anaesthesia was studied. The purpose of this work was to evaluate the efficacy of 2-CP for ambulatory surgery of short duration with this combination of blocks. Twenty-five patients ASA 1 and 2 were studied. The techniques used have been described by Winnie and Labat. A total dose of 60 ml of 2-CP 3% was injected. Latency time before anaesthesia was approximately seven min for the lumbar plexus and 14 min for the sciatic nerve. The mean duration of the motor block was 71 +/- 16 min, 76.8 +/- 15.3 min and 75 +/- 12.7 min for the sciatic, femoral and obturator nerves respectively. The mean duration of the sensory block was 88.7 +/- 20.9 min, 83.3 +/- 16.4, 79.7 +/- 17.8 min and 93.7 +/- 22 min for the sciatic, femoral, obturator and lateral femoral cutaneous nerves respectively. Success rate was 92% and no major complication occurred. We conclude that a combination of the lumbar plexus and the sciatic nerve blocks with 2-CP is a useful technique for ambulatory surgery of short duration.

Adult↗
PubMed1994

Anatomy of the posterior approach to the lumbar plexus block.

The purpose of this study was to describe the relation of the lumbar plexus with the psoas major and with the superficial and deep landmarks close to it. Four cadavers were dissected and 22 computed tomography files of the lumbosacral region studied. Cadaver dissections demonstrated that the lumbar plexus, at the level of L5, is within the substance of the psoas major muscle rather than between this muscle and the quadratus lumborum. The femoral nerve lies between the lateral femoral cutaneous and obturator nerves. However, while the lateral femoral cutaneous nerve is in the same fascial plane as the femoral nerve, the obturator nerve can be found in the same plane as the two other nerves or in its own muscular fold. Radiological data provided the following measurements: the femoral nerve is at a depth of 9.01 +/- 2.43 cm; the psoas major medial border is at 2.73 +/- 0.64 cm from the median sagittal plane; and its lateral border is at 6.41 +/- 1.61 cm from the same plane. It is concluded that the lumbar plexus is within the psoas major, that the obturator nerve localization within the psoas major varies and that computed tomography data define precisely the relationship of the lumbar plexus with superficial and deep landmarks.

Adipose Tissue↗
PubMed1994

Posterior approach to the lumbar plexus combined with a sciatic nerve block using lidocaine.

A combination of lumbar plexus block, by a posterior technique, and sciatic nerve block can be a useful technique for outpatient anaesthesia. The purpose of this study was to examine the clinical characteristics of these blocks using lidocaine and to measure the serum lidocaine concentrations. Forty-five patients, undergoing lower extremity surgery, were studied. Sciatic nerve and lumbar plexus blocks were made with lidocaine, 680 mg with adrenaline 0.3 mg. For each patient the following data were collected: weight, age, sex, site of surgery, time to perform each block, needle depth, speed of onset of the sensory and motor blocks in the territories of the sciatic, femoral, obturator and lateral cutaneous (sensory) nerves and postoperative analgesic requirements. Lidocaine serum concentrations were measured in ten of these patients at 0, 2, 5, 10, 30, 60, 90 and 120 min after the second block. Analgesia was complete in 88% (40/45) of the patients. The remaining five patients needed analgesics (fentanyl 150 micrograms or less). Despite the high dose of lidocaine, the serum concentrations were within safe limits (mean +/- SD) (CMAX = 3.66 +/- 2.21 micrograms.ml-1). Only one patient had a serum concentration > 5 micrograms.ml-1 (CMAX = 9.54 micrograms.ml-1). This was associated with a contra-lateral extension of the block. We conclude that this combination of blocks is a valuable alternative for unilateral lower extremity anaesthesia. However, clinicians must be aware of the implications of a contra-lateral extension of the block.

Adolescent↗