Biomedical subjects

F Nau

Publications and source records attributed to F Nau.

At least 19 recordsLinked to original sources

PubMed2005

Hen egg white fractionation by ion-exchange chromatography.

Major hen egg white proteins have been widely studied for their functional properties but these studies still are unable to explain, alone, all of the biological properties of hen egg white. Hence, it is still interesting to produce pure and non-altered proteins to improve our knowledge on the biological properties of hen egg white. Presently, identification and characterization of both bioactive peptides and minor proteins from hen egg white is essential work for progressing in the understanding of hen egg white biological properties. With this objective in mind, a new process for a complete "mucin free" hen egg white fractionation based on ion exchange chromatography is proposed. "Mucin free" egg white is fractionated into six different fractions. Four of them are high-recovery yield purified fractions of lysozyme, ovotransferrin, ovalbumin and flavoprotein. The two other fractions are enriched in recently detected minor proteins in hen egg white.

Animals↗

PubMed2003

Cloning and characterization of HEP21, a new member of the uPAR/Ly6 protein superfamily predominantly expressed in hen egg white.

Using two-dimensional (2D)-PAGE, partial protein internal sequencing, and PCR with degenerate primers, we cloned a novel cDNA named HEP21 from hen egg white. The 0.5-kb cDNA encodes a 106 amino acid protein with a cysteine spacing pattern suggesting that HEP21 is a new member of the uPAR/CD59/Ly-6/ snake neurotoxin superfamily. The closest homology of HEP21 is to mouse Ly-6C. Unlike most members of this protein family, HEP21 is not glycosylphosphatidylinositol (GPI)-anchored but is a secreted protein, as indicated by its localization and the presence of a signal peptide in its sequence. Moreover, HEP21 appears as an original member of this protein superfamily because it is predominantly expressed in a tissue, i.e., the oviduct, and especially the magnum where the egg white components are secreted.

Amino Acid Sequence↗

PubMed2001

Comparison of different electrophoretic separations of hen egg white proteins.

The hen egg white protein composition has not yet been fully defined. To improve the knowledge of this biological fluid, the most usual and recently developed electrophoretic methods have been used: SDS-PAGE, native-PAGE, isoelectric focusing (IEF), and 2-dimensional electrophoresis (2DE). Seven of the major known proteins were thus identified in at least one electrophoretic system. Isoforms of ovotransferrin, ovalbumin, and ovomucoid were visualized when pI was used for the separation. Two-dimensional electrophoresis allowed separation of a very large number of spots. In each of the four systems, some components were revealed but not identified, and unknown spots were particularly numerous with 2DE. With this technique, many spots corresponding to small acidic proteins were highlighted, among which was the Ch21 protein, whose presence in hen egg white was thus confirmed. This study thus constitutes, to our knowledge, the first proteomic investigation of hen egg white.

Animals↗

PubMed2000

Simple rapid procedure for preparation of large quantities of ovalbumin.

A simple rapid procedure for preparation of large quantities of highly purified homogeneous ovalbumin from egg white by using an anion exchanger is described. It is based on the principle of frontal chromatography. The volume of "mucin-free" egg white loaded onto the column was determined in order to exceed resin capacity. Thus, competition between proteins for resin sites was created. Owing to its high negative charge density, ovalbumin drives other egg white proteins from the column progressively. Two hundred and fifty milliliters of Q-Sepharose FF gel eluted isocratically with 0. 5 M NaCl extracted 9.55 g of ovalbumin with a purity rate of 83%. A 6.75 g amount of ovalbumin, with a purity rate of 94%, was recovered with an isocratic elution program using 0.14 M NaCl. Purified ovalbumins were compared by electrophoresis and analytical chromatography with other ovalbumin preparations.

Chromatography, Ion Exchange↗

PubMed1999

High intensity pulsed electric fields applied to egg white: effect on Salmonella Enteritidis inactivation and protein denaturation.

High-intensity electric fields have been successfully applied to the destruction of Salmonella Enteritidis in diaultrafiltered egg white. The effects of electric field strength (from 20 to 35 kV x cm(-1)), pulse frequency (from 100 to 900 Hz), pulse number (from 2 to 8), temperature (from 4 to 30 degrees C), pH (from 7 to 9), and inoculum size (from 10(3) to 10(7) CFU x ml(-1)) were tested through a multifactorial experimental design. Experimental results indicate that, for Salmonella inactivation, the electric field intensity is the dominant factor with a strongly positive effect, strengthened by its positive interaction with pulse number. Pulse number, temperature, and pH have also significant positive effects but to a lesser extent. In the most efficient conditions, the pulsed electric field (PEF) treatment is capable of 3.5 log10 reduction in viable salmonellae. Simultaneously, the measure of surface hydrophobicity does not indicate any increase after PEF treatment. These results suggest that no protein denaturation occurs, unlike what is observed after comparable heat treatment in terms of Salmonella inactivation (55 degrees C for 15 min).

Colony Count, Microbial↗

PubMed1998

cDNA cloning, structural organization, and expression of the sheep NRAMP1 gene.

Mouse resistance to several intracellular pathogens including Mycobacteria, Leishmania, and Salmonella is under the control of the Chromosome (Chr) 1 Natural Resistance Associated Macrophage Protein I gene (Nramp1). This gene could have an economic and health importance for domestic animals and humans as well. Therefore, equivalents of the NRAMP1 gene have been cloned by several research groups in various animal species. To study in sheep the influence of the NRAMP1 gene on the susceptibility to intracellular pathogens induced diseases, we have cloned the sheep NRAMP1 cDNA by screening a splenic cDNA library. The genomic organization of the sheep NRAMP1 gene was then determined by sequencing the exon/intron boundaries. The transcription start points (tsp) from the NRAMP1 mRNA have been located with primer extension experiments. RT-PCR reactions have been used to determine the profile of mRNA expression of this gene.

Amino Acid Sequence↗

PubMed1997

Genomic organization of the sheep immunoglobulin JH segments and their contribution to heavy chain variable region diversity.

The sheep immunoglobulin heavy chain Igh-J locus has been characterized in order to determine the genomic organization of JH segments and their contribution to heavy chain diversity. The locus contains six segments, of which two are functional and four are apparently pseudogenes. These segments span a 1.8 kilobase (kb) region. The distance between JH-ps4 (the 3'-most segment) and the first domain of the mu-chain encoding constant gene is about 5 kb. The two functional JH segments have a standard upstream recombination signal sequence, including heptamer and nonamer sequences separated by a 22-23 nucleotide spacer, and end with a RNA donor splice site. These two segments possess all the characteristic JH invariant residues and are found in expressed mu heavy chain variable regions. The 5' functional JH1 segment is used in more than 90% of the cDNAs sequenced to date. The contribution of JH segment germline multiplicity to variable regions diversity appears therefore to be minimal. Comparison with other mammalian JH segments shows that all loci are very closely related and probably have evolved from a common ancestral locus.

Amino Acid Sequence↗

PubMed1996

The sheep Ig variable region repertoire consists of a single VH family.

Nine germ-line Ig heavy chain variable (VH) segments (including three pseudogenes) were isolated from a genomic DNA library, and the other six were obtained by PCR, using 5'and 3' primers deduced from the first three. They appear to belong to a homogeneous VH gene family, with >80% sequence identity. This sheep VH gene family is related to the human VH4 family and to the murine VH1 subgroup (clan II). Southern blot analysis shows a maximum of 10 positive restriction fragments; therefore, the nine VH genes isolated probably constitute the major part of the repertoire. Thirty-one expressed mu variable regions (and one gamma 1 variable region) were obtained from adult spleen by either cDNA cloning or anchored reverse transcriptase-PCR; they are >80% similar to each other (in their leader to framework 3 regions) and to the germ-line sequences as well. The sheep VH repertoire thus seems to derive from a small (approximately 10 members) germ-line gene family, and its diversification must rely chiefly on junctional (D and/or N regions) diversity and somatic hypermutations.

Animals↗

PubMed1992

Primary structure of a monoclonal kappa chain in myeloma with light chain deposition disease.

Previous data suggest that structural abnormalities of immunoglobulin light chains may be responsible for non-amyloid light chain deposition disease (LCDD). We report on the complete primary sequence deduced from complementary (c)DNA analysis of a normal-sized kappa chain in a case of myeloma-associated LCDD. The patient's urine contained a kappa type Bence-Jones protein made of monomers and dimers of an unglycosylated kappa chain. The bone marrow myeloma cells contained intracellular kappa and gamma chains by immunofluorescence. Biosynthesis experiments showed the production of normal-sized gamma chains and of kappa chains with the same apparent molecular mass (Mr) in SDS gels as the urinary kappa chain (26,000-27,000). These kappa chains were secreted as assembled IgG molecules and as a large excess of free monomers and dimers. The complete sequence of two identical cDNA clones derived from a normal-sized kappa messenger RNA indicated that this kappa chain belonged to the rare V kappa IV subgroup. The kappa mRNA had an overall normal structure made up of the V kappa IV sequence rearranged to J kappa 1 and followed by a normal constant exon of the Km(3) allotype. The variable region differed from the V kappa IV-J kappa 1 germline sequence by 17 amino acid substitutions. The peculiar sequence of the variable region of this kappa chain of a rare subgroup might relate to its tissue deposition.

Aged↗

PubMed1990

Alternate splice sites within the human VH gene coding sequences lead to truncated Ig mu-chains.

The Burkitt's lymphoma cell lines JBL2 and Ly91 contain intracytoplasmic mu chains without L chain by immunofluorescence. These mu chains and the corresponding cDNA are abnormally short. Detailed analysis of the productive H chain alleles showed that the cell lines JBL2 and Ly91 use a VHIV and a VHIII subgroup gene, respectively. In both cell lines, the 3' part of the V exon was markedly abnormal and contained several stop codons. Point mutations affected the JH6 donor splice site in the JBL2 gene, precluding normal splicing into C mu. Although the JH2 splice site was unmodified in Ly91, it was not used. As in JBL2, splicing occurred at an alternate splice site present 5' of these alterations, thus removing all these abnormalities from the mature transcripts. This alternate splicing explains the presence in both cell lines of truncated mRNA and proteins lacking two-thirds of the V region.

Base Sequence↗

PubMed1990

Immunoglobulin light chain transcripts with altered V regions in Burkitt's lymphoma cell lines producing short mu chains.

The Burkitt lymphoma cell lines Ly91, Ly47 and Ly66 produce short mu chains in the absence of associated light (L) chains. mu mRNA lack two-thirds of the variable (V) region due to an alternate splicing event that takes place within V genes. L chain genes were found to be rearranged in these lines and to lead to transcripts with altered V regions. Although these abnormal transcripts displayed specific abnormalities in each cell line, they shared some features, including large deletions involving the V-J junction. In one cell line, L chain transcripts made up of either V kappa I subgroup gene J kappa 3 or V kappa IV gene-J kappa 4 recombination products bore a similar alteration: the donor splice site of the rearranged J kappa segment was modified, leading to the alternate use of the donor site of either the leader peptide exon or the next 3' unrearranged J kappa segment.

Base Sequence↗

PubMed1990

Production of an abnormal mu chain with a shortened VHIV subgroup variable region in a Burkitt's lymphoma cell line.

The Burkitt's lymphoma cell line Ly66 produces a short mu chain which lacks 4 kDa in apparent molecular mass. Study of the corresponding messenger RNA showed it to be 0.3 kb shorter than normal mu transcripts. The cDNA sequence of the mu transcripts began by a short VH region consisting of the first one-third of a VHIV subgroup gene segment. It was followed by a normal mu constant region. This VH region coded for 38 amino acids, thus differing from two truncated VHIV regions previously reported in other Burkitt's lymphoma cell lines, which were interrupted at codon +26 by an alternate splicing event. In addition, the Ly66 variable sequence bore several point mutations and contained two potential N-glycosylation sites.

Amino Acid Sequence↗

PubMed1988

Burkitt's lymphoma cell lines producing truncated mu immunoglobulin heavy chains lacking part of the variable region.

Burkitt's lymphoma cell lines Ly91, Ly47 and JBL2 contained intracytoplasmic mu chains without light chains and variably expressed membrane mu chains. All three cell lines produced similarly truncated heavy chains with an apparent molecular mass of 64 kDa instead of the normal 74 kDa. Both the intracellular and secreted forms of the mu chains were similarly truncated. The membrane and the secreted forms of mu mRNAs were shorter than normal. The cDNAs contained a leader sequence, followed by about one third of a variable region (VH), then by the constant region (C mu). Interruption of the variable region took place near the end of the framework region 1 (FR 1) in Ly 47 and JBL2, and within the complementarity determining region 1 (CDR1) in Ly91. In the three lines, the joining between VH and C mu maintained the normal open reading frame throughout the variable and constant regions.

Amino Acid Sequence↗

PubMed1987

[Behavior therapy in functional syndromes].

Twenty patients presenting functional syndromes of various kinds, localizations and severities without any tangible anatomic-morphological causes having been found, were treated on the basis of behavioural self-direction. Self-rating instruments were used for therapy control. Following a mean 45-61 sessions of individual therapy, highly significant reductions were stated in respect of the probable occurrence of the usually extensive symptoms. This positive outcome was found to persist at follow-up twelve months later. This management approach turned out a very economical one. Further investigations appear indicated.

Behavior Therapy↗

PubMed1983

A study of some molecular and kinetic properties of two tRNA methyltransferases from mouse plasmocytoma.

A tRNA(adenine-1)methyltransferase and a tRNA(cytosine-5)methyltransferase have been partially purified from mouse plasmocytoma MOPC 173. Their apparent Mr are 200000-230000 and 110000-140000, respectively, as determined by gel filtration and density gradient centrifugation. Both enzymes exhibit maximum activity in the presence of high concentrations of monovalent cations (0.175 M and 0.25 M KCl, respectively) and in the absence of magnesium. Their kinetic constants have been determined at various KCl concentrations, with several tRNA species as substrates. These constants may differ by more than one order of magnitude, depending upon the substrate used, and they are strongly dependent upon the ionic concentration as well. The possibility that the tRNA(adenine-1)methyltransferase from mouse plasmocytoma is different from the homologous enzyme purified from a normal rat tissue [Glick, J. M. and Leboy, P. S. (1977) J. Biol. Chem. 252, 4790-4795] is discussed.

Animals↗

PubMed1981

tRNA methylation. A rapid and simple method for determination of total radioactivity and methylated base distribution in the same sample.

A simple and rapid method is described for the analysis of in vitro methylated tRNA. The total radioactivity is first determined after trichloroacetic acid-precipitation and filtration. Then the tRNA is hydrolyzed while still on the filter, and the bases are separated by thin-layer chromatography. This method eliminates the need for duplicate sample preparation, and has proved especially useful when the amount of tRNA and/or tRNA methylase is limited.

Carbon Radioisotopes↗

PubMed1981

Discrepant effects of unilateral and bilateral forebrain lesions on the visual performance of pigeons.

The monocular and binocular performance of pigeons with bilateral, unilateral or sham lesions in the telencephalic Wulst was tested with visual discrimination tasks. Unilateral lesions yielded a marked deficit when the animals could only use the eye contralateral to the lesion. Otherwise the accomplishments of the ablated animals did not differ from that of the controls. The reciprocal inhibition of symmetrical visual brain stem centers is thought to have been unbalanced through the one-sided interruption of a known pathway descending from the Wulst.

Animals↗

PubMed1980

Intracellular distribution of antibodies in rat lymph node cells during primary response. A kinetic study.

The kinetics of appearance of immunoglobulin- and antibody-containing cells in rat lymph nodes was studied by immunoenzymatic techniques between days 12 and 90 after a single injection of horse-radish peroxidase. Three distinct phases appeared during the primary repsonse. Between days 8 and 17, essentially no antibody-containing cells were detectable, while non-specific immunoglobulin-containing cells amounted to up to 9% of the total cell population. The peak of antibody-forming cells was observed between days 18 and 24, and their number decreased slowly during the last phase. Days 18-20 were marked by a sharp peak of a distinct category of cells containing antibodies in a limited area of their cytoplasm. One day before this event, the percentage of immunoglobulin-forming cells decreased drastically, and remained very low throughout the response. These results are discussed on the basis of a previously published model, which proposes that non-specific immunoglobulin-containing cells are precursors to antibody-containing cells, and that the transformation between these two categories of cells involves a stage where antibody synthesis is restricted to a few endoplasmic reticulum vesicles.

Animals↗