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T Baron

Publications and source records attributed to T Baron.

At least 37 records · Page 2Linked to original sources
PubMed2000

A comparative study of immunohistochemical methods for detecting abnormal prion protein with monoclonal and polyclonal antibodies.

Transmissible spongiform encephalopathies are associated with the accumulation of abnormal prion protein (PrP(Sc)) in the central nervous system which can be detected immunohistochemically. Using a monoclonal antibody (L42) to an epitope on the first alpha-helix of ruminant PrP, we compared previously reported immunohistochemical antigen unmasking and "visualization" systems. In addition, a variety of polyclonal and monoclonal antibodies to other epitopes on ruminant PrP were assessed. Antigen unmasking by hydrated autoclaving and proteinase K treatments, and antigen detection with L42 and an avidin-biotin complex system, enabled intra- and extra-neuronal PrP(Sc)to be demonstrated in scrapie-affected sheep carrying three different PrP alleles, as well as in cases of bovine spongiform encephalopathy.

Amino Acid Sequence↗
PubMed1999

Overexpression and purification of an immunologically reactive His-BIV capsid fusion protein.

The gene of the capsid protein of bovine immunodeficiency virus (BIV) was linked to a sequence encoding for six histidines and expressed as the (His)6 p26 capsid fusion protein. The fusion protein was strongly expressed as both soluble and insoluble forms after induction by isopropylthio-beta-d-galactoside. Purification was based on interaction of the hexa-histidine polypeptide with metal ions. Expression could represent 11% of the total protein in Escherichia coli, allowing more than 20 mg of highly purified protein to be obtained per liter of bacterial culture. The (His)6 p26 capsid fusion protein purified by immobilized metal affinity chromatography reacted specifically in Western blot with sera from cattle experimentally infected by BIV, as well as with two monoclonal antibodies directed against different epitopes of the Gag protein. The ease of expression, purification, and specificity of this fusion protein should permit a thorough study of prevalence of BIV infection in large-scale serological studies of field samples.

Animals↗
PubMed1998

Detection of bovine immunodeficiency-like virus infection in experimentally infected calves.

Detection of BIV virus infection by serological means, PCR and virus isolation in experimentally infected calves is described. Viral sequences were specifically detected by PCR in peripheral blood mononuclear cells (PBMCs), with primer systems located in the gag, pol and tat regions of the viral genome. An enzyme-linked oligosorbent assay (ELOSA) in microtiter plates is described, for the detection of PCR products, the sensitivity of which was shown to be comparable to that of membrane hybridization detection. Serological response of the animals against the BIV p26 protein was shown, using a recombinant fusion protein ((His)6p26) expressed in E. coli and purified by metal affinity chromatography, in ELISA and Western blot studies. The presence of infectious virus was demonstrated by its rescue, by virus isolation in cell cultures, from PBMCs during a one year follow-up.

Animals↗
PubMed1998

Sensitivity of the Western blot detection of prion protein PrPres in natural sheep scrapie.

The sensitivity of Western blot detection of PrPres using two different extraction procedures on brain material of 30 scrapie-affected sheep was compared. Whereas PrPres could be detected in all sheep after extraction with the first method, 30% did not give any signal after extraction with the second method. However, the second method, when positive, permitted the detection of PrPres from smaller amounts of infected brain tissue. When used with the ruminant specific monoclonal antibody p4, the second method gave positive signals corresponding to less than 12.5 microg of scrapie-infected brain, that, up to now, is the highest sensitivity described for PrPres detection from naturally infected ruminant brains. The overall results showed highly variable levels of PrPres between sheep and are presented in relation to breed, survival time and animal genotype data. Further progress can thus be expected for PrPres detection in prion diseases, if more efficient extraction procedures and more sensitive immunological reagents are used. Such technical improvements could contribute to more accurate diagnosis in animals affected naturally.

Animals↗
PubMed1997

Biochemical properties of protease resistant prion protein PrPsc in natural sheep scrapie.

Prions are infectious agents involved in neurodegenerative diseases, such as scrapie in sheep and goats, bovine spongiform encephalopathy (BSE) in cows and Creutzfeldt-Jakob disease (CJD) in humans. These pathogens are characterized by unusual properties, and, in particular, by their strong resistance to common procedures of disinfection used against conventional microorganisms. A major component of highly infectious fractions is a proteinase K-resistant prion protein PrPsc (PrP-res), the normal host prion protein PrPc being sensitive to PK (PrP-sen). We used a biochemical approach to further characterize PrPsc protein in natural sheep scrapie. Western blot analyses using rabbit antiserum that recognized both normal and pathologic sheep prion proteins, were undertaken to study the biochemical behaviour of PrPsc extracted from brains of sheep naturally infected with scrapie after protease digestion and under denaturing conditions. Increasing concentrations of urea (1-7 M) or GdnSCN (0.25-3 M) and different pH from 2 to 11 were tested for their effects on protease resistance of PrPsc. Alkaline pH (pH = 10) and high concentrations of urea (> 3 M) and GdnSCN (> 0.75 M) greatly decreased the protease resistance of the prion protein. Identical experiments carried out on three different sheep from the same flock gave similar results. The biochemical behaviour of PrPsc under denaturing conditions and in the presence of proteinase K could thus provide a biochemical means for further characterization of different natural scrapie isolates.

Animals↗
PubMed1996

Serologic evidence for bovine immunodeficiency virus infection in France.

We report herein on the first serologic detection of antibodies to bovine immunodeficiency virus (BIV) in France. Serum samples from dairy and beef cattle from southwestern and western France (Landes and Vendée) were tested using a western blot assay with a recombinant 53 kDa gag precursor derived from the Louisiana BIV R29 isolate. We performed our study on the oldest animals from 37 different herds that were under serologic follow up for previous bovine leukemia virus infection. Overall, 398 selected bovine sera were assayed and 15 serum samples from 8 herds reacted with the recombinant 53 kDa BIV R29 gag. Interestingly, reactions obtained with French sera were weaker than with positive Louisiana sera, a finding that may indicate the occurrence of distinct French and Louisiana BIV variants.

Animals↗
PubMed1995

The bovine immunodeficiency-like virus (BIV) is transcriptionally active in experimentally infected calves.

We have studied the infection by the bovine immunodeficiency-like virus (BIV) in three experimentally infected calves, by polymerase chain reaction (PCR) and reverse transcription-polymerase chain reaction (RT-PCR), from the peripheral blood mononuclear cells (PBMC). Two primer pairs located in the gag and pol regions of the viral genome allowed to detect the viral genomic DNA by PCR, as well as the unspliced genomic viral RNA transcript, by RT-PCR. We also present the evidence of the presence in peripheral blood mononuclear cells (PBMCs) of a mRNA transcript of the regulatory trans-activator tat gene, according to the splicing pattern of the viral genome, by use of reverse transcription followed by nested PCR. The active expression of the virus in these animals was further assessed by the sequential rescue of the virus from unstimulated PBMCs in cell culture, from 4 weeks until 15 months following the infection.

Animals↗
PubMed1995

[Bovine spongiform encephalopathy].

A new Transmissible Sub-acute Spongiform Encephalopathy (TSSE) appeared in 1986 in the bovine species, mainly concerning Great Britain where more than 100,000 cases have now been identified. This review describes the features of the disease that showed it belonged to this particular group of diseases. The currently available hypothesis about the origin of the disease, in Great Britain as well as in other countries where it has also been recognized, are presented. Lastly, the potential risks for the future, in the bovine species but also for other animal species or man, are discussed, in light of the known experimental data.

Animals↗
PubMed1994

Long term in vitro growth of lymphoblastoid cells in the dog.

Attempts to grow in vitro cells from canine lymphoid organs led to observation of the occurrence of a spontaneous, delayed and transitory growth of cells in suspension in two lymphoma cultures (out of 20) but also from one (out of 20) dog with unrelated disorder. These cells, with a lymphoblastoid cytological appearance, lack detectable immunoglobulins in cases studied originating from lymphomatous dogs, and have a pseudodiploid canine karyotype. Neither appearing fully immortalized nor showing any tumorigenicity in nude mice or immunosuppressed newborn rats, these may be of lymphoblastoid nature, rather than arising from some tumorous cells. The mechanisms that have triggered the cell growth are still unknown, but such cells could remind of human lymphoblastoid cells, associated with lymphotrophic viruses, still unknown in the dog.

Animals↗
PubMed1992

Report on the first outbreaks of the porcine reproductive and respiratory syndrome (PRRS) in France. Diagnosis and viral isolation.

We describe the first known occurrence in France of the porcine reproductive and respiratory syndrome (PRRS), a new porcine disease that first appeared in Germany in November 1990. Outbreaks of the disease appeared in November 1991 in Brittany (France), with comparable clinical features to those observed in other countries, were definitively confirmed by a serological analysis from the affected animals using an immunoperoxidase monolayer assay with PRRS-virus infected alveolar macrophages. Furthermore, we report the isolation from one serum of a filtrable agent that, in view of its cytopathic effect for porcine alveolar macrophages and of the serologic reactivity of the infected cells, appears similar to the Lelystad virus that has been implicated in the etiology of the disease.

Animals↗
PubMed1992

An enzyme linked immunosorbent assay (ELISA) for the detection of antibodies to the porcine reproductive and respiratory syndrome (PRRS) virus.

An indirect enzyme-linked immunosorbent assay (ELISA) has been developed for the rapid detection of antibodies to the porcine reproductive and respiratory syndrome (PRRS) virus in pig sera. Compared to the immunoperoxidase monolayer assay (IPMA) which was the only test available up till recently for serodiagnosis of the disease, the ELISA test proved to be more sensitive, particularly for early detection of antibodies. The test was also highly specific for the PRRS virus inasmuch as it scored negative all the hyperimmune sera directed to other swine viruses and the field sera sampled before the outbreak of the disease in Brittany (France). This easy test now provides further possibilities for epidemiological surveys and could also be a reliable tool for new sanitary prophylaxis of the disease.

Animals↗