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Biomedical subjects

J Frey

Publications and source records attributed to J Frey.

At least 199 records · Page 11Linked to original sources
PubMed1991

Use of antibodies against the P36 protein of Mycoplasma hyopneumoniae for the identification of M. hyopneumoniae strains.

Mycoplasma hyopneumoniae, the principal aetiological agent of porcine enzootic pneumonia, synthesizes a 36 kDa protein (P36) which is an early and strong immunogenic factor in experimentally and naturally infected swine. Polyclonal antibodies were made against the recombinant P36 protein in rabbits and used for the identification of M. hyopneumoniae by the immunoblot technique. The proteins from the M. hyopneumoniae reference strains and from 13 M. hyopneumoniae field strains isolated from naturally infected pigs in Switzerland, Hungary, France and Canada were analysed by the immunoblot technique using anti-P36 antibodies. All 13 field strains and the three reference J strains of M. hyopneumoniae, received from different collections and laboratories, exhibited a strong reaction with a protein of 36 kDa indicating that the P36 protein is a common M. hyopneumoniae antigen. None of the different porcine Mycoplasma species including M. flocculare, M. hyorhinis, M. hyosynoviae, A. axanthum, A. laidlawii and A. granularum showed any reaction on the immunoblot with the anti-P36 antibodies. In addition, we have found no reaction with anti-P36 antibodies using 47 different Mycoplasma or Acholeplasma species isolated from human, mice, rat, poultry, ruminant, dog and cat. In conclusion we have shown that P36 is a protein that is a common antigen of M. hyopneumoniae strains and is not found in other Mycoplasma or Acholeplasma species tested. Because of its high specificity, P36 protein, or antibodies made against this protein can be used for the identification of M. hyopneumoniae strains.

Acholeplasma↗
PubMed1991

Cloning and expression of a species-specific early immunogenic 36-kilodalton protein of Mycoplasma hyopneumoniae in Escherichia coli.

Mycoplasma hyopneumoniae, the etiologic agent of porcine enzootic pneumonia, synthesizes a 36-kDa protein which is an early and strong immunogenic factor in experimentally and naturally infected swine. The gene encoding this protein was cloned by screening a gene library of M. hyopneumoniae DNA with rabbit hyperimmune serum made against whole M. hyopneumoniae cells and convalescent-phase swine serum. Analysis of the recombinant protein expressed in Escherichia coli by immunoblot techniques showed that the protein is expressed in E. coli in its full length and does not cross-react with proteins from M. flocculare or M. hyorhinis. Genetic analysis showed that the gene was expressed from the lac promoter of the vector and seems to be translationally initiated from its own ribosome binding site. Subcloning in a transcriptional fusion vector to optimize expression resulted in production of the 36-kDa protein in E. coli at levels up to 30% of total protein.

Animals↗
PubMed1991

Nucleotide sequence of the hemolysin I gene from Actinobacillus pleuropneumoniae.

The DNA sequence of the gene encoding the structural protein of hemolysin I (HlyI) of Actinobacillus pleuropneumoniae serotype 1 strain 4074 was analyzed. The nucleotide sequence shows a 3,072-bp reading frame encoding a protein of 1,023 amino acids with a calculated molecular size of 110.1 kDa. This corresponds to the HlyI protein, which has an apparent molecular size on sodium dodecyl sulfate gels of 105 kDa. The structure of the protein derived from the DNA sequence shows three hydrophobic regions in the N-terminal part of the protein, 13 glycine-rich domains in the second half of the protein, and a hydrophilic C-terminal area, all of which are typical of the cytotoxins of the RTX (repeats in the structural toxin) toxin family. The derived amino acid sequence of HlyI shows 42% homology with the hemolysin of A. pleuropneumoniae serotype 5, 41% homology with the leukotoxin of Pasteurella haemolytica, and 56% homology with the Escherichia coli alpha-hemolysin. The 13 glycine-rich repeats and three hydrophobic areas of the HlyI sequence show more similarity to the E. coli alpha-hemolysin than to either the A. pleuropneumoniae serotype 5 hemolysin or the leukotoxin (while the last two are more similar to each other). Two types of RTX hemolysins therefore seem to be present in A. pleuropneumoniae, one (HlyI) resembling the alpha-hemolysin and a second more closely related to the leukotoxin. Ca(2+)-binding experiments using HlyI and recombinant A. pleuropneumoniae prohemolysin (HlyIA) that was produced in E. coli shows that HlyI binds 45Ca2+, probably because of the 13 glycine-rich repeated domains. Activation of the prohemolysin is not required for Ca2+ binding.

Actinobacillus↗
PubMed1991

Early treatment of ischemic stroke with a calcium antagonist.

We performed a feasibility and safety study (phase II) of nicardipine, a calcium antagonist, in 57 patients. The objectives of the study were to begin therapy as early as possible (less than or equal to 12 hours) after the onset of ischemic stroke and to administer as high a dose as possible. All patients received an intravenous infusion of nicardipine for 72 hours, starting with a dose of 3 mg/hr and increasing to a maximum dose of 7 mg/hr. Upward titration of the dose was limited by a 10% decrease in blood pressure or a 20 beats/min increase in pulse. Intravenous therapy was followed by 30 days of oral therapy. The mean +/- SD interval from onset of stroke to commencement of therapy was 9.1 +/- 5.4 hours. Adverse reactions consisted primarily of hypotension requiring discontinuation of therapy in four patients. Score on a graded neurologic examination increased from 41/100 at baseline to 64/100 at 3 months for the 41 patients completing follow-up. There was no correlation between the dose of nicardipine administered and outcome, but the 11 patients starting therapy less than or equal to 6 hours after onset did better than those starting therapy 6-12 hours after onset. Further study of very early therapy with nicardipine is justified.

Blood Pressure↗
PubMed1991

[Lysine and collagen].

The amount of lysine in collagen is only 3 or 4% of total aminoacids, but it has an important function in the constitution of the cross-links between the molecules to built the fibrils and the fibers of collagen. For this function, some lysine molecules must be hydroxylated and other oxidized to aldehyde compounds. Some deficits in these metabolic pathways are responsible for heritable diseases of the connectivite tissue as types IV and IX of the Ehlers-Danlos syndrome, lathyrism, the Menkes kinky hair syndrome, Cutis-Laxa or the type II of osteogenesis imperfecta. The contribution to fibrosis is also discussed.

Collagen↗
PubMed1991

Endocytosis of human IgG:Fc receptor complexes by transfected BHK cells.

We have analyzed the mode of uptake of human beta FcRII molecules expressed in BHK cells (clone 2/14). When challenged with aggregated human IgG (ahIgG), these cells bind the ligand at 4 degrees C and endocytose the IgG: receptor complexes rapidly upon warming to 37 degrees C, as seen by fluorescence microscopy with antibodies directed against human IgG. Using 125I-labeled ahIgG, we found that 40% of the bound ligand was internalized within 15 min, and approximately 60% within 2 h. Surface replication and thin sectioning combined with immunogold labeling revealed that the ligand was taken up by coated vesicles and was transferred to the endosomal/lysosomal compartment. This was confirmed by confocal laser microscopy of cells double labeled for clathrin and ahIgG. After modulation of the coated vesicle pattern by hypertonic medium, ahIgG transport was impaired. These data show that a single isoform of human FcRII, expressed in an animal cell negative for Fc receptors, can use the coated vesicle based endocytic pathway of the host cell. Reincubation of cycloheximide-treated cells with a second batch of ligand showed that approximately 20% of the beta FcRII was recycled. This finding is in apparent contrast to the fate of the endogenous Fc receptors expressed on mouse macrophages.

Animals↗
PubMed1990

[The role of intraoperative angioplasty performed simultaneously with reconstructive vascular surgery].

21 arterial lesions were dilated intraoperatively in conjunction with primary vascular reconstructive procedures. Of these, five involved the iliac vessels, fourteen the femoropopliteal segment, one the subclavian artery and one the common carotid artery. 19 intraoperative balloon dilatations were successful and two were unsuccessful. Only one catheter-related complication were noted, it was a perforation of the wall of the artery caused by guidewire. No deaths occurred as a result of the combined procedure. Attention is drawn to the advantage of the intraoperative balloon dilatation and the authors recommend it on adequate conditions and correct indication.

Adult↗
PubMed1990

Distribution, inducibility and biological function of the cloned and expressed human beta Fc receptor II.

A cDNA encoding the human beta Fc gamma receptor II (FcRII) was isolated from a placental cDNA library. Analysis of the predicted amino acid sequence indicates that this receptor is synthesized with a 42-amino acid leader sequence. The mature protein consists of 249 amino acids. The leader sequence and the cytoplasmic domain are strikingly different from the CDw32 antigen but show great homology to the mouse beta 2FcR. RNA blot analysis of human cells using CDw32 and beta FcRII-specific DNA fragments demonstrated one beta FcRII transcript (1.7 kb) in B cells and in HL-60 cells which were induced to differentiate along a monocyte-macrophage pathway by phorbol 12-myristate 13-acetate treatment. Under these conditions the CDw32 transcripts (2.5 and 1.7 kb) are induced to a minor extent in HL-60 cells. In contrast, the 2.5-kb CDw32 transcript is strongly induced in HL-60 cells which have been induced to differentiate into granulocytes by exposure to dimethylsulfoxide. To determine the biological properties of the beta FcRII, we expressed the antigen in FcR- hamster cells. Only immune complexes but not monomeric human IgG were bound significantly. Bound ligand was efficiently internalized within 15 min and it was then found in vesicular structures. Thus the low-affinity beta FcRII is able to internalize ligands without cooperation with any other FcR.

Amino Acid Sequence↗
PubMed1990

Organization of human FcRII and FcRII-like (beta FcRII) genes: structural homology to HLA class I and class II genes.

Genomic EMBL 3 DNA clones representing part of the human Fc gamma receptor II and the beta Fc gamma receptor II genes were characterized. One of them contains the first five exons including the 5' flanking region of the beta FcRII gene. The signal peptide, the extracellular domains, the putative membrane spanning region, and the first amino acids of the cytoplasmic region encoded by these five exons are spread over approximately 11 kb. Another genomic DNA clone comprises four exons encoding the second extracellular domain and the transmembrane and cytoplasmic regions of the FcRII. Alignment of the genomic DNA clones reveals that these FcR genes are identically organized. Comparison of the corresponding regions of these clones shows that not only the exons are strikingly homologous but also the splice junctions and parts of the intervening sequences are conserved. Furthermore, the genomic organization of FcRII and HLA-class I resemble each other.

Antigens, Differentiation↗
PubMed1990

[Postoperative fever].

When faced with a case of postoperative fever, the diagnostic procedure is of paramount importance. It's first step is analytical and clinical, basing on examination of the patient and search for indicators of gravity. The first aim of examination is to recognize a septic collection related to surgery and/or intensive therapy. The following step is synthetical and has to consider the type of surgery and the status of the patient. The symptomatic therapy of high fever relies on opioids and non-steroidal anti-inflammatory agents; their indications and limits are discussed.

Anti-Bacterial Agents↗
PubMed1990

Isolation of the Actinobacillus pleuropneumoniae haemolysin gene and the activation and secretion of the prohaemolysin by the HlyC, HlyB and HlyD proteins of Escherichia coli.

The gene encoding the c. 105 kD secreted haemolysin protein of the porcine pathogen Actinobacillus pleuropneumoniae serotype 1 has been isolated by screening a lambda gt11 expression library in Escherichia coli with antiserum raised against the wild-type protein. A derivative recombinant DNA pJFF702 expressed the hlylA haemolysin gene from the pUC19 lac promoter but the resulting haemolysin I protein remained within the E. coli cell and was haemolytically inactive. Export of the intracellular A. pleuropneumoniae prohaemolysin out into the medium was achieved by the presence in trans of the E. coli haemolysin secretion genes hlyB and hlyD, and high levels of intracellular haemolytic activity were attained similarly by the E. coli post-translational haemolysin activator gene, hlyC. Southern hybridization of A. pleuropneumoniae parental DNA nevertheless indicated only a low degree of nucleotide sequence identity to the haemolysin structural and secretion genes hlyA and hlyB of E. coli. The data show that despite substantial nucleotide sequence divergence the A. pleuropneumoniae serotype 1 haemolysin determinant is closely related to that which is dispersed throughout other Gram-negative human and animal pathogens.

Actinobacillus↗
PubMed1990

Hemolysin patterns of Actinobacillus pleuropneumoniae.

The secreted hemolytic activities produced by the reference strains and field isolates of the 12 serotypes and 2 subtypes of Actinobacillus pleuropneumoniae were analyzed. Serotype 1 produced a Ca2(+)-inducible hemolysin, which was previously characterized as a 105-kilodalton protein and was named hemolysin I (HlyI). Serotypes 2, 4, 6, 7, and 8 produced a different hemolytic activity that was not inducible by Ca2+ but required this ion for its activity. The hemolytic activity produced by these serotypes was much weaker than that found in serotype 1 and was not neutralized by rabbit antibodies against HlyI. It was, however, neutralized by serum from pigs that were experimentally infected with a serotype 2 strain and was called hemolysin II (HlyII). Serotypes 5a, 5b, 9, 10, and 11 produced both HlyI and HlyII. In these strains, HlyI was the major contributor to the hemolytic activity. The remaining serotypes, 3 and 12, produced a very weak hemolytic activity, which was not further analyzed. Immunoblot analysis of the culture supernatants from all 12 serotypes with rabbit polyclonal antibodies directed against HlyI revealed reactions with a protein in the 105-kilodalton size range for all serotypes, indicating that HlyI and HlyII might be serologically related. Strains producing active HlyI seem to belong to serotypes that are generally considered to be virulent types and that are frequently isolated from pigs in severe pleuropneumonia outbreaks.

Actinobacillus↗
PubMed1990

[Long time hospitalization in pediatric surgical intensive care units].

If the great majority of children stay in intensive care unit only for few days, a small group of patients stay for a long period of time, varying from 3 months to five years or even more. The small group include two very different entities. On the one hand, newborns with a congenital malformation, often digestive e.a. with a short small intestine, need a long time parenteral nutrition. In such cases, the family disturbance, the opening of the unit to the parents, the awakening of the infant, his psychomotor and affective attainments are as many problems for the whole medical team. On the other hand, children severely injured in accidents, that occurred on public thoroughfares or in domestic house holds, most of the time are older than 3 years. These children may present important central neurologic sequelae, or other major sequelae (multiple trauma, burn injuries, hypoxic cervical spinal cord injury). Their education, activities, games, and family relations must be actively supported. Does the hospital structure, and particularly the intensive care unit, meet such requirements in very long time hospitalization?

Adolescent↗
PubMed1990

[Behavior on de-epidermized human skin of unspecified reconstituted collagen tissue].

The aim of our study was to assess the behaviour of a despecified collagen tissue laid on desepidermized human skin (taking area of skin graft). The collagen tissue was prepared according to Bell's method. The collagen was latticed, contracted by fibroblasts, chemically treated (formaldehyde or glutaraldehyde) to stabilize the fibrils and then despecified by cialit treatment. This tissue laid on superficial, non infected wounds very rapidly took a necrotic appearance and was totally lysed after 10 to 12 days without any modification of the healing course. No bacterial infection was observed. Histological and ultrastructural studies showed desorganization of collagen fibre bundles and tissue invasion by inflammatory cells. Circulating antibodies to collagen were absent at day 30. This model lacks interest as a substitute for superficial tissue replacement, but a healing function could be assigned to its high chemotactic power for polynuclears and macrophages and should allow its use in cases of deep tissue loss.

Aged↗