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Biomedical subjects

M Girard

Publications and source records attributed to M Girard.

At least 307 records · Page 17Linked to original sources

[Automated management of the Center of Transfusion Blood Bank equipped with Autogrouper 16 C].

We describe in this paper a first processing of direct connection and data management from the Autogrouper 16 C. It bas been used since November 1980 in the Centre Départemental de Tranfusion Sanguine des Hauts-de-Seine. The automation of validation procedures, updating blood donors' file and labelling blood units allows the suppression of clerical errors that are the major cause of Trannsfusion accidents. The progressive development of programming lets a good adaptation of working methods and makes the setting of the system much easier in the Blood Bank.

Antibodies↗

Molecular cloning of the genome of poliovirus type 1.

Poliovirus cDNA.RNA hybrids were prepared from the Mahoney strain of poliovirus type 1 by using reverse transcriptase (RNA-dependent DNA nucleotidyltransferase) and cloned in the Escherichia coli plasmid pBR322. Bacteria colonies carrying recombinant plasmids were selected by in situ hybridization with virus-specific RNase T1-resistant oligonucleotides. Analysis of the cDNA inserts by restriction mapping and electron microscopy showed that the cloned cDNAs, the longest of which was 3.2 kilobase pairs, originated from various parts of the viral RNA, covering at least 99% of the genome length. Due to overlapping of the clones, the restriction map of the poliovirus genome could be reconstructed. The complete 5' end of the genome was successfully cloned in at least one of the recombinant plasmids, pPV1-366.

Cloning, Molecular↗

Protein kinases associated with the adenovirus single-stranded DNA-binding protein.

Protein kinase activities copurifying with the 72000-Mr DNA-binding protein of adenovirus on DNA-cellulose chromatography and gel filtration in acrylamide/agarose have been partially characterized and purified. One of these kinases was found to phosphorylate efficiently the viral DNA-binding protein in vitro and to be stimulated severalfold by the addition of histones, protamine, or polyamines. The kinase does not, however, phosphorylate histones, protamine, casein, or phosvitin. A second protein kinase was also recovered from single-stranded DNA-cellulose which is able to phosphorylate the 72000-Mr DNA-binding protein, but which is inhibited by the addition of histones. Phosphorylation in vitro of the 72000-Mr DNA-binding protein from the ts125 mutants of adenovirus by the histone-stimulated protein kinase was found to be thermosensitive.

Adenoviridae↗

Restriction map of poliovirus type 2 cDNA.

Poliovirus type 1 RNA was reverse-transcribed into c-DNA and inserted at the Pst I site of the plasmid vector pBR322 of E. coli. Resulting recombinant plasmids were analyzed by hybridization with RNase T1-resistant 32P-labeled oligonucleotides, and by restriction enzyme mapping. All of the genome was cloned in a series overlapping cDNA inserts, the longest of which was 3.2 kb. The restriction map of the poliovirus cDNA is presented.

Avian Myeloblastosis Virus↗

[Determination of Lewis and P blood group phenotypes with the aid of the Autogrouper 16C].

For more than one year, we are using the Autogrouper 16 C, at the beginning the prototype machine according to Rechsteiner, for ABO, Rhesus and Kell determinations and now Lewis and P grouping increases its possibilities, using free chanels. The connection to a PDP 11 computer enables us to sophisticate the interpretation of results and to use, for example, unadsorbed reagents.

Autoanalysis↗

Characterization of a new adenovirus type 5 assembly intermediate.

Adenovirus H5 (Ad5) DNA-protein complexes were extracted with ammonium sulphate (0.2 M) from virus-infected HeLa cell nuclei at 18 h after infection. Analysis of the material by centrifugation through discontinuous sucrose gradients in heavy water revealed the existence of several populations of molecules which were identified, in order of increasing buoyant density, as mature DNA-protein complexes, replication complexes, assembly intermediates and virions. When observed under the electron microscope, some of the assembly intermediates showed a capsid with a tail of entirely double-stranded (ds) DNA, or of dsDNA continued by a portion of single-stranded (ss) DNA thickened by a coat of E-72 K DNA binding protein. Singly or doubly-forked Ad5 replicating DNA molecules partially packaged in virus capsids were also observed. It is suggested that these molecules could be assembly intermediates, i.e. one of the first steps of assembly corresponding to virus DNA entering pre-formed capsids or their precursors. The fact that replication was still going on at one end of many of the DNA molecules in the intermediates, while encapsidation was taking place at the other, raises the possibility of a coupled DNA replication-packaging process in the formation of adenovirions.

Adenoviruses, Human↗

The influence of fluoro-phenyl-alanine on the synthesis of simian virus 40 DNA and T antigens.

Treatment of Simian Virus 40 (SV40) infected monkey cells with fluorophenylalanine (FPA) resulted in increased uptake of thymidine by the cells, and progressive inhibition of both viral and cellular DNA synthesis. Viral DNA synthesis was more sensitive to inhibition by FPA than cell DNA synthesis. Synthesis of SV40 T antigens was however unaffected by FPA, as judged from immunofluorescence assays. The M.W. of the major polypetides immunoprecipitated from cell extracts by antibodies from tumor bearing hamster sera was similarly unaffected. It is suggested that T antigen synthesized in the presence of FPA is non functional.

Animals↗

Characterization of simian virus 40 transcriptional intermediates in infected CV1 cell nuclei.

Simian Virus (SV40) transcriptional intermediates (T.I.) were isolated from infected cell nuclei incubated in vitro in the presence of the four ribonucleoside triphosphates. The nascent mRNA strands in the viral DNA-RNA hybrid molecules were hydrogen bonded to their template by 200-250 nucleotides on the average, as judged from the extent of their RNase resistance and the aspect of T.I. under electron microscope after treatment with 50 per cent formamide. The RNA polymerase involved (RNA polymerase II) synthesized up to full length transcripts at a rate of approximately 150 nucleotides/min. at 25 degrees C. Each SV40 infected cell was found to contain about 200 active T.I. molecules at the peak of late transcription. The DNA in the T.I. molecules was exclusively form I DNA only in cell infected with the tsA30 mutant of SV40 that had been transferred to non-permissive temperature in order to arrest DNA replication, but both form I DNA and molecules behaving as replicative intermediates (R.I.) in wild type infected cells.

Animals↗

[Use of the BG 15 autoanalyzer for automatic recording and determination of ABO, Rhesus and Kell blood groups].

The use of the 15 channels blood grouping autoanalyzer with automated recording and interpretation proved its fiability through more than 30 000 tests for ABO, Rh and Kell determinations. Misreactions come mostly from the carry-over phenomenom. However no discrepancy was noticed for ABO determination. Only few ones occured for Rhesus and Kell, leading to false positive interpretations which were always corrected by checking the event recorder track. New improvements are expected from the automatic phasing using a microprocessor and further extents are setting up with the connection to a computer.

ABO Blood-Group System↗

Storage of proteins in the rough endoplasmic reticulum of human hepatocytes in a patient with normal blood proteins, on oral contraceptives.

Aspects of protein storage in the rough endoplasmic reticulum of hepatocytes, comparable with those reported in alpha1-antitrypsine (AAT) deficiency, have been observed in the course of jaundice in a woman presenting no evident abnormality in AAT or other blood proteins. In light microscopy, most hepatocytes contained characteristic globular inclusions but they were PAS negative and did not react with anti-AAT antibodies. This storage of protein ceased at the time the jaundice disappeared. Prolonged treatment with high doses of contraceptive steroids may have been involved in this peculiar reaction of the hepatocytes.

Adult↗

Selective inhibition of in vitro DNA synthesis dependent on phiX174 compared with fd DNA. I. Protein requirements for selective inhibition.

Crude extracts of Escherichia coli selectively convert fd viral DNA and not phiX174 DNA to duplex DNA via a complex series of reactions one of which involves RNA polymerase. Reactions leading to formation of fd duplex-replicative (RFII) structures have been reconstituted with purified proteins from E. coli. Maximal synthesis requires the combined action of E. coli binding protein, DNA elongation factor I, DNA elongation factor II preparations (which are a mixture of dna Z and DNA elongation factor III), DNA polymerase III, DNA-dependent RNA polymerase, Mg2+, dATP, dGTP, dCTP, dTTP, and ATP, GTP, CTP, and UTP. In contrast to crude extracts of E. coli, purified protein fractions do not distinguish between fd DNA and phiX174 DNA in duplex DNA formation. The addition of crude fractions of E. coli to the purified components listed above selectively permits fd RFII formation and prevents phiX RFII formation. This selective inhibition was used as an assay to isolate proteins essential for this phenomenon; they include RNase H, discriminatory factor alpha, and discriminatory factor beta.

Adenosine Triphosphatases↗

DNA polymerase activities in growing cells infected with simian virus 40.

Growing CV1 cells were infected with simian virus 40 (SV40), and the levels of DNA polymerases-alpha, -beta, and -gamma were analyzed in the cytoplasm, nuclear Triton wash, and nucleus. In the cytoplasmic fraction, the amount of alpha-, beta-, or gamma-polymerase remained unaltered after SV40 infection. The activity of DNA polymerase-alpha increased five- to sixfold in the nuclear Triton wash and threefold in the nuclei and then remained enhanced only inside the nuclei. That of DNA polymerases-beta and gamma increased mostly in the nuclei after infection. These results suggest that DNA polymerase-alpha could be the major enzyme involved in SV40 DNA replication.

Cell Line↗