[Primary tuberculous infection and spontaneous pneumothorax].
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Humans↗
Biomedical subjects
M Girard
Publications and source records attributed to M Girard.
Use of capillary electrophoresis for the characterization of human serum albumin heterogeneity.
Human serum albumin (HSA) is used in large amounts as an excipient in many biopharmaceutical formulation to prevent loss of the active ingredient through adsorption and/or degradation. Traditionally, iso-electric focusing has been used to demonstrate charge heterogeneity in HSA preparations. In an effort to develop new methods for the analysis of formulation components, a capillary zone electrophoresis method was developed for the analysis of HSA. Under initial separation conditions using untreated silica capillaries and 20 mM sodium phosphate, pH 6.0 as electrophoretic buffer, HSA migrated as a single peak. Addition of 1,4-diaminobutane allowed separation of several components which could be further resolved by varying the buffer pH. Optimal separation conditions were attained at 5 mM 1,4-diaminobutane and pH 8.5. The reproducibility of the separation conditions was verified by using capillaries from a different manufacturer. A comparative analysis of HSA preparations from different manufacturers provided evidence that the method may be used to qualitatively differentiate individual preparations. The analysis of rhEPO formulations, composed largely of HSA, showed levels of heterogeneity comparable to that of HSA preparations. Electrospray ionisation mass spectrometry (ESA-MS) was used as an independent method to confirm the heterogeneous nature of HSA.
Electrophoresis, Capillary↗
An aid to correct positioning of the ProSeal laryngeal mask.
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Anesthesiology↗
Prevalence of hepatitis B virus infection in the Austral archipelago, French Polynesia: identification of transmission patterns for the formulation of immunization strategies.
A sero-epidemiological survey of hepatitis B virus (HBV) infection in a randomly selected sample of 957 persons from the population of the Austral Island group in French Polynesia was conducted as a first step before developing an immunization programme strategy. Prevalence rates of HBsAg ranged from 3.09% to 27% in the different islands of the group with a weighted mean of 10.48%, while the prevalence rate for at least one marker ranged from 46.91% to 81.03% with a weighted mean of 64.12%. In the 0-11 months and 1-4 years age groups, 2.08% and 10.57%, respectively, of the children were HBsAg carriers. These findings, when compared to the mean population carrier rate of 10.48%, suggest that HBV transmission occurred mostly after the first year of life. The highest prevalence rate for HBeAg positivity was in the 5-19 years age group (more than 40% of the HBsAg carriers were HBeAg positive), suggesting that contagiousness was greatest in childhood and adolescence. HBsAg was found in 11.45% of women of child-bearing age and HBeAg in 19.09% of women positive for HBsAg. It is concluded that immunization of newborns and infants, using vaccine alone, should be the most effective strategy for reducing HBV infection in the Austral Islands archipelago.
Age Factors↗
Reliability of data on haematotoxicity of antidepressants. A retrospective assessment of haematological monitoring in clinical studies on tricyclics.
At the present time, there is some concern over the haematological adverse effects of antidepressants. The present paper examines retrospectively 314 clinical trials in depression which were published in English or French between 1958 and 1986 and which used at least one of the following tricyclics: amitriptyline, clomipramine, imipramine. Two decision tables assess the depth of detail with which haematological monitoring and safety were described by the authors of each study. The results indicate that more than 98% of the papers under scrutiny either provide no information on the subject or do so in a manner which is entirely subjective. Thus, it seems that current knowledge of haematotoxicity of antidepressants is almost entirely the result of spontaneous reports from doctors, with its well-known drawbacks.
Amitriptyline↗
Expression of a poliovirus neutralization epitope at the surface of recombinant bacteria: first immunization results.
We devised a procedure to construct strains of Escherichia coli which expose at their surface a foreign antigen genetically inserted into LamB, an outer membrane protein. In particular, we showed that amino acid residues 93-103 of poliovirus type 1 capsid polypeptide VP1, which correspond to the C3 neutralization epitope, when inserted into two different external loops of LamB (after residues 153 and 374 of the mature protein), yielded the synthesis of stable hybrid proteins named, respectively, 153-C3 and 374-C3. The poliovirus epitope was accessible to monoclonal antibody C3 at the cell surface. In the present work, these two hybrid proteins were injected into rabbits by the intravenous route in the form of live recombinant bacteria, and the humoral response to the poliovirus epitope was studied. With construction 153-C3, the subcutaneous route was also assayed using solubilized hybrid protein. The C3 viral sequence inserted in the two different regions of LamB were found to be immunogenic. Different types of antibodies specific to the C3 peptide were raised with the two construction: anti-peptide and antiviral particle antibodies. These first results indicate that the LamB presentation vector system constitutes a mode of peptide coupling which may lead to the elaboration of a new type of live vaccine.
Animals↗
Episiotomy and faecal incontinence.
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Anal Canal↗
Expression of the yellow fever virus envelope protein using hybrid SV40/yellow fever viruses.
The cDNA coding for the yellow fever virus (YFV) envelope protein (E) was inserted into an SV40 vector under the control of the late promoter in place of the VP1 gene. The recombinant virus expressed a 52-Kd polypeptide which was detected by immunoprecipitation with a monoclonal antibody raised against the E protein. Surprisingly, this protein was visualized in the nucleus of the infected cells. The possible presence of a sequence involved in nuclear migration of the E protein and naturally ignored during virus infection is discussed. The sequence coding for the mature E protein is directly preceded in the YFV genome by a sequence of 45 nucleotides coding for a 15-amino-acid long hydrophobic oligopeptide. The sequence of this oligopeptide was inserted into the SV40 recombinant virus between the ATG codon and the first codon for the E protein. The E protein expressed by this new SV40 recombinant virus was found to be localized in the cytoplasm of the infected cells in association with intracellular membranes. These results strongly suggest that the 15-amino-acid long hydrophobic region naturally plays a role as a signal peptide for the E protein.
Animals↗
Construction of a chimaeric type 1/type 2 poliovirus by genetic recombination.
A chimaeric poliovirus carrying a type-2-specific neutralization epitope on a type 1 capsid was created by site-directed mutagenesis of the Mahoney strain of poliovirus type 1. An EcoRV and a HindIII restriction sites were first constructed in the cDNA of poliovirus type 1 at nucleotide positions 2756 and 2786, respectively, i.e. on either side of the sequence encoding neutralization epitope C3 (VP1 amino acids 93-103), which is part of neutralization site NImI. The cDNA sequence framed by the two sites was next taken out and replaced by custom-made oligonucleotides encoding the equivalent region of VP1 from the Lansing strain of poliovirus type 2. The DNA from the plasmid carrying such a hybrid construct was transfected onto CV1 cells generating a chimaeric virus, v510. Neutralization of v510 with a panel of monoclonal antibodies showed that v510 has lost the poliovirus type 1 C3 epitope but acquired a new, poliovirus type-2-specific neutralization epitope. Preliminary results indicate that v510 also shows neurovirulence for mice, which is a specific trait of the Lansing strain of poliovirus type 2.
Antibodies, Monoclonal↗
[cRNA probes (riboprobes) synthesized in vitro for detecting enterovirus by molecular hybridization].
Radioactively labelled RNA transcripts made in vitro of various fragments from cDNA clones of poliovirus type 1 and of hepatitis A virus under the control of bacteriophage T7 or SP6 promoters have been evaluated for diagnostic purposes. The RNA transcripts were 2 orders of magnitude more sensitive as hybridization probes than corresponding cDNA preparations labelled by the nick translation procedure. A combination of hybridization analysis and sequence comparison showed that some regions of the genome of a number of enteroviruses are highly conserved, while others show very little homology; the general order of conservation is: 5'-non-coding greater than 3'-terminal greater than central (2C) greater than VP3 greater than VP1. The 350 bases of the poliovirus VP1 region were highly specific for that virus, while the 450-bases of the 5'NC region showed extensive cross-reaction with other enteroviruses. However, these probes did not hybridize with HAV, which was detected only by HAV-specific riboprobes. The transcripts have been successfully applied as hybridization probes in diagnostic tests on supernatants of infected cell culture lysates and in clinical samples, mainly in stool extracts.
Animals↗
Ability of linear and cyclic peptides of neutralization antigenic site 1 of poliovirus type 1 to induce virus cross-reactive and neutralizing antibodies.
Eight peptides encompassing neutralization antigenic site 1 of poliovirus type 1 (residues 93-103 of VP1) were synthesized in linear or cyclized form and used to immunize rabbits. The resulting anti-peptide antibodies were tested for their ability to react with linear peptide 95-104, with infectious virus D-particles and heated C-particles and for their capacity to neutralize poliovirus infectivity. A good correlation was observed between the ability of different peptide antisera to immunoprecipitate D-particles and neutralize virus infectivity. The peptides that induced a neutralizing antibody response in the highest number of immunized animals contained flanking residues 104-115 in addition to the 93-103 residues of the epitope. However, a high neutralizing antibody titre was also obtained in two of ten animals immunized with peptide 93-104 cyclized via an amide bond between Asp93 and Lys103. It seems, therefore, that, at least in rabbits, the T-cell epitope recently identified in residues 103-115 of VP1 need not be present in the peptide immunogen in order to obtain poliovirus-specific neutralizing antibodies.
Amino Acid Sequence↗
Preliminary characterization of cis interactions between mutations of poliovirus genome.
The effects of three distinct point mutations in the P3 genomic region of poliovirus type 1 (Mahoney strain) have previously been analysed. A U to C change at position 5658, which modifies the proteinase 3C, is responsible for a small plaque phenotype and defective RNA replication. An A to G change at position 7256, which modifies the replicase 3D, is responsible for temperature sensitivity. A C to U change at position 7348, which is silent at the protein level, has no apparent phenotypic effect. These three mutations were introduced into a single poliovirus genome by recombinant cDNA techniques. The resulting plurimutant virus, vHA507, was temperature-sensitive and exhibited a small plaque phenotype. Both these characters were accentuated compared to those of the viruses in which the mutations were on separate genomes. Spontaneous ts+ revertants of vHA507 arose by true reversion of the replicase mutation. The small plaque phenotype and defective RNA replication were partially suppressed in the case of these revertants, despite the fact that the proteinase mutation was still present on their genomes. This suppression was assumed to be due to the presence of the silent mutation at position 7348. This system provides a simple model of cis interactions within the poliovirus genome.
DNA-Directed RNA Polymerases↗
Study of the immunogenicity of different recombinant Mengo viruses expressing HIV1 and SIV epitopes.
Recombinant Mengo viruses expressing heterologous genes have proven to be safe and immunogenic in both mice and primates, and to be able to induce both humoral and cellular immune responses (Altmeyer et al., 1995, 1996). Several recombinant Mengo viruses expressing either a large region (aa 65-206) of the HIV1 nef gene product, or cytotoxic T lymphocyte (CTL) epitopic regions from the SIV Gag (aa 182-190), Nef (aa 155-178) and Pol (aa 587-601) gene products were engineered. The heterologous antigens were expressed either as fusion proteins with the Mengo virus leader (L) protein, or in cleaved form through autocatalytic cleavage by the foot-and-mouth disease virus 2A protein. Rhesus macaques and BALB/c mice inoculated with the Mengo virus SIV recombinants failed to develop CTL responses against the SIV gene products, while one of the HIV-Nef recombinants induced a weak CTL response in mice directed to an HIV1 Nef peptide spanning positions 182-198. In contrast, BALB/c mice immunized with vaccinia virus recombinants expressing HIV1 Nef developed a strong CTL response to the 182-198 peptide and also responded to a second peptide spanning positions 73-81. These results indicate that Mengo virus recombinants expressing HIV1 Nef and SIV CTL epitopes are weak immunogens. One of the fusion recombinants expressing SIV CTL epitopes failed to infect macaques even when used at high doses, while the recombinant expressing HIV1 Nef as a fusion protein failed to infect BALB/c mice. These results demonstrate that the expression of certain heterologous sequences as fusion proteins with L can result in the loss of the ability of the recombinant to infect normally susceptible animals.
Amino Acid Sequence↗
Patient-controlled epidural analgesia with fentanyl-bupivacaine: influence of prior dural puncture.
BACKGROUND AND OBJECTIVES: Combined spinal epidural anesthesia (CSEA) involves the epidural administration of local anesthetic and opioid solutions adjacent to the prior dural puncture, potentially increasing their diffusion into the subarachnoid space. This study was designed to evaluate the influence of dural puncture on the adequacy and extent of analgesia, and drugs requirements of patient-controlled epidural analgesia (PCEA) in the postoperative period. METHODS: In this prospective double-blind study, 40 patients undergoing major abdominal surgery under general anesthesia followed with PCEA were randomly assigned to either group I (preoperative insertion of an epidural catheter) or group II (preoperative dural puncture with a 25-g Quincke needle + insertion of an epidural catheter). Postoperatively, a PCEA pump delivered an infusion of 0.1% bupivacaine + fentanyl (3 microg/mL) at 5 mL/h. Participants were allowed to self-administer 5-mL boluses of the same solution with a 15-minute lock-out interval. Hourly epidural solution requirements were recorded for 40 hours. Sensory and motor block, and pain scores were also analyzed. RESULTS: There was no difference between groups with regard to epidural solution requirements, pain scores, spread of sensory blockade, or intensity of motor block. CONCLUSION: Dural puncture with a 25-gauge Quincke needle, performed as part of CSEA, does not influence the drug requirements when a combination of 0.1% bupivacaine and fentanyl (3 microg/mL) is used for PCEA after major abdominal surgery.
Abdomen↗
HIV F/3' orf encodes a phosphorylated GTP-binding protein resembling an oncogene product.
Apart from the retroviral gag, pol and env the HIV genome contains the F (3' orf) gene which encodes a polypeptide of 206 amino acids which is myristylated at the N-terminal and whose function is unknown. We have expressed the F gene in Escherichia coli and from a recombinant vaccinia virus, VVTGfHIV. The F-protein produced in VVTGfHIV-infected mammalian cells is myristilated, and is phosphorylated by protein kinase C at a residue close to the N-terminus like pp60-src (ref. 5). Purified bacterial F-protein also shows the GTPase, autophosphorylation and GTP-binding activities reported for the ras gene product. Furthermore, we show that expression of F in a CD4+ cell line down-regulates the CD4(T4) antigen. These results suggest that F is important in the pathophysiology of AIDS (acquired immune deficiency syndrome).
Antigens, Differentiation, T-Lymphocyte↗
Evaluation of a new blood typing instrument.
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Blood Grouping and Crossmatching↗
Inhibition of herpes simplex virus multiplication by 2,3-bis(acetyl mercaptomethyl)-quinoxalin.
The drug 2,3-bis(acetyl mercaptomethyl)-quinoxalin (BAMMQ) was 99.9% inhibitory for herpes simplex virus (HSV) multiplication in cell cultures at concentrations of 1.6 X 10(-5) M or less. The drug was not inactivating for the virus, did not interfere with adsorption and penetration of the virus, and was still active when added as late as 12 h after infection with HSV. BAMMQ did not prevent HSV DNA replication, but interfered with a late stage of virus assembly and/or maturation.
Cell Line↗
[Methodological obstacles encountered in the evaluation of psychotherapies of schizophrenic patients. A general review].
Through a critical review of literature, authors suggest a methodological reflexion about problems in evaluation of effects of psychotherapies of schizophrenics, taking into account not only individual techniques but also institutional ones with psychotherapeutic orientations. Among the main difficulties are: evaluation with too much unspecific and over simplified criteria, for example criterium of rehospitalisation; lack of real comparability between groups, or techniques, and lack of validity of randomisation, still considered as a pawn of methodological rigour; diversity of levels of experience, and education of therapists, too unmatched model of functional organisation of the different centers, and in the same time too unequal recruitment of patients and at length of proposed therapeutic programs. Moreover, and even before these methodological bias, a more basic obstacle must be considered. It is the non-opening of models to the falsification, denounced by Bignami as the impossible verification of the heuristic value of the proposed models, ie the impossible verification of the utility of the proposed programs for individual therapies of schizophrenics.
Bias↗