PubMed Health⌕ Search

Biomedical subjects

R Alain

Publications and source records attributed to R Alain.

At least 19 recordsLinked to original sources
PubMed2000

Mechanical and structural properties of milk protein edible films cross-linked by heating and gamma-irradiation.

The mechanical properties of cross-linked edible films based on calcium caseinate and two type of whey proteins (commercial and isolate) were investigated. Cross-linking of the proteins was carried out using thermal and radiative treatments. Size-exclusion chromatography performed on the cross-linked proteins showed that gamma-irradiation increased the molecular weight of calcium caseinate, while it changed little for the whey proteins. However, heating of the whey protein solution induced cross-linking. For both cross-linked proteins, the molecular weight distribution was >/=2 x 10(3) kDa. Combined thermal and radiative treatments were applied to protein formulations with various ratios of calcium caseinate and whey proteins. Whey protein isolate could replace up to 50% of calcium caseinate without decreasing the puncture strength of the films. Films based on commercial whey protein and calcium caseinate were weaker than those containing whey protein isolate. Electron microscopy showed that the mechanical characteristics of these films are closely related to their microstructures.

Gamma Rays↗
PubMed1998

Characterizations of natural and induced polyhedrin gene mutants of Bombyx mori cytoplasmic polyhedrosis viruses.

Bombyx mori cytoplasmic polyhedrosis virus (BmCPV) polyhedrin gene from natural mutant strain B, B2 and P was cloned and the amino acid sequence was compared with that of wild-type virus (strain H). Chimeric polyhedrin genes (8 types) containing only one site of mutation within the amino acid sequence were also constructed. Fifteen types of polyhedrin genes were introduced into a baculovirus expression vector and hexahedral, acicular, pyramidal, or amorphous polyhedra were formed in infected cells. These results demonstrated that the shape of polyhedra as well as the crystallization pattern of the polyhedrin could be changed by mutations at respectively N-terminal and C-terminal regions of BmCPV polyhedrin gene.

Animals↗
PubMed1995

Ultrastructural characteristics and morphogenesis of porcine reproductive and respiratory syndrome virus propagated in the highly permissive MARC-145 cell clone.

A Québec reference strain of PRRSV (IAF-KLOP) was successfully propagated in MARC-145 cells, a highly permissive cell clone to PRRSV derived from the MA-104 cell line. Purified extracellular virions appeared as pleomorphic but mostly spherical enveloped particles, 50-72 nm in diameter, with an isometric core about 25-30 nm. By indirect immunofluorescence, detection of viral antigens within the cytoplasm was possible as soon as 6 h.p.i. Nucleocapsids, budding at smooth endoplasmic reticulum (ER), and enveloped viral particles that tended to accumulate in the lumen of ER or Golgi vesicles, were the main features of the viral morphogenesis. The virus apparently was released by exocytosis.

Animals↗
PubMed1990

Growth of poliovirus using alginate entrapped-anchorage dependent Vero cells.

Obligatory anchorage dependant Vero cells were successfully grown in gelatinous-like macrocarriers made of calcium alginate. Entrapped single cells were immobilized within the polymerized alginate matrix divide to form large spherical clumps of cells. A cell density of 17 x 10(6) cells/ml of alginate with over 95% viability was obtained after 14 days in spinner flasks. When subjected to poliovirus type I infection, spherical masses of Vero cells progressively showed extensive cytopathic effect but remained entrapped in the alginate matrix of the macrocarriers. Virus was released into a cellular debris free-like supernatant and reached a peak titer of 10(8.0) TCID50/ml after 72 hours.

Alginates↗
PubMed1990

Electron microscopy of mouse thymic virus.

Thymuses of mice infected with mouse thymic virus (MTV), a herpesvirus, were examined with the electron microscope. Damage was manifested by distortions of cells, nuclei, and other organelles, with cellular aggregation apparent at advanced stages of necrosis. Infected cells also contained MTV particles and filaments. Negative staining showed that naked MTV capsids were icosahedral in shape with diameters ranging from 95 to 110 nm. Enveloped capsids appeared spherical and ranged in diameter from 125 to 165 nm. Immunogold labeling revealed the presence of antigenic sites around both naked and enveloped capsids, and also on the 10 nm filaments. The use of electron microscopy and immunohistochemical techniques as diagnostic tools to demonstrate MTV infection could prove valuable for the detection of the infection and the study of its pathogenesis.

Animals↗
PubMed1989

Comparison of caprine, human and bovine strains of respiratory syncytial virus.

A new continuous ovine kidney cell line allowing the growth of caprine, human and bovine respiratory syncytial virus was used to minimize host cell related variations for the direct comparison of the viral ultrastructures, serological relationships and structural protein profiles. Results show that all three strains are closely related although a closer relationship was found between bovine and caprine RS.

Animals↗
PubMed1989

A new bacteriophage of Corynebacterium glutamicum isolated from swine waste.

A bacteriophage for Corynebacterium glutamicum strain LP-6 was isolated from swine waste. It belongs to the Siphoviridae family or Bradley morphologic group B, has a narrow host range, and is sensitive to chloroform and resistant to carbon tetrachloride. The phage is unstable (96% inactivation) in swine waste stored for 4 months at 22 C. The DNA has a molecular weight of approximately 20 Md, cohesive ends, and numerous restriction endonuclease sites. The phage differs from other known C glutamicum phages.

Animals↗
PubMed1988

Rapid titration of bovine, caprine and human RS virus by a micro-immunoperoxidase assay using a monoclonal antibody and a permissive ovine kidney cell line.

An indirect immunoperoxidase micro-assay, using a continuous cell line derived from ovine kidney cells (OK) and a previously characterized monoclonal antibody (7C2), specific for an exposed and highly conserved epitope of the fusion protein of different strains of RS virus, was used advantageously to rapidly titrate bovine, caprine and human strains of RSV by either quantal (TCID50) or plaque forming assays. Virus titers, obtained in less than 36 h, were in agreement with those obtained by the conventional plaque assays which required an incubation period of 4 days or more. This assay is also applicable to micro-neutralization of fusion inhibition assays for testing serum or screening monoclonal antibodies.

Animals↗
PubMed1988

Further biological, serological and biochemical characterization of North American, European and Southeast Asian strains of bovine herpesvirus 1 compared with other alphaherpesvirinae members.

Hemagglutination activity, structural protein profiles and neutralization assays were used in a comparative study of bovine herpesvirus 1 strains from the U.S.A., Canada, Great Britain, Denmark and Malaysia with equine, feline and human herpesviruses in order to further characterize the bovine herpesvirus 1 hemagglutinin. Bovine herpesvirus 1 strains of different geographical origins all showed hemagglutinating activity for mouse erythrocytes; furthermore, feline herpesvirus 1 was also shown to hemagglutinate mouse erythrocytes. Analyses of partly purified viruses showed that a distinctive and specific polypeptides profile is associated with each species of herpesviruses used in our study; strains of bovine herpesvirus 1 from North America, Europe and Southeast Asia however, presented a remarkable similarity as to their electrophoretic protein patterns. A protein similar to the 97-kDa bovine viral hemagglutinin was not identified with the hemagglutinating feline herpesvirus. An important neutralization epitope on the bovine viral hemagglutinin was also not found on feline, equine and human herpesviruses but was identified on all bovine strains tested from North America, Europe and Southeast Asia stressing the importance of the bovine hemagglutinin for eventual prophylactic purposes.

Animals↗
PubMed1988

Electron microscopic evidence for bridges between bovine respiratory syncytial virus particles.

Electron microscopic examination of ultrathin sections of a continuous cell line of ovine kidney (OK) origin, infected by bovine respiratory syncytial virus (BRSV), revealed the presence of well defined bridges between virus particles. This is the first report of this novel structure. Observation of ultrathin sections of human RSV Long strain also grown on OK cells did not show inter-particle bridges and therefore suggested that this structure could be specific to BRSV. The biological significance of these bridges is not clear at this time; a possibility is that the bridges are formed by the fusion protein of BRSV which is known to cause cell fusion. Besides the structural implications, the importance is in relation to purification strategies for this virus, which must now take into account that most of the viral particles occur in large aggregates.

Animals↗
PubMed1987

Rapid virus subunit visualization by direct sedimentation of samples on electron microscope grids.

Airfuge direct ultracentrifugation of viral samples on electron microscope grids offers a rapid way for concentrating viral particles or subunits to facilitate their detection and study. Using the A-100 fixed angle rotor (30 degrees) with a K factor of 19 at maximum speed (95,000 rpm), samples up to 240 microliters can be prepared for electron microscopy observation in a few minutes: observation time is decreased and structural details are highlighted. Using latex spheres to calculate the increase in sensitivity compared to the inverted drop procedure, we obtained a 10- to 40-fold increase in sensitivity depending on the size of particles. Application of this technique to rubella virus permitted better visualization of viral membrane subunits on the particles. Rubella hemagglutinin immuno-stimulating complexes preparations were also better visualized and their morphology conserved after direct ultracentrifugation on the specimen grids. Similar observations are reported for respiratory syncytial virus associated subunits.

Animals↗
PubMed1985

An enteric coronavirus of the rabbit: detection by immunoelectron microscopy and identification of structural polypeptides.

The immunoelectron microscopy (IEM) technique has been used for the detection of a rabbit enteric coronavirus (RECV). Immune serum was prepared in guinea pigs; the viral antigen used for the immunization procedure was obtained from the caecum of a sick rabbit, concentrated by centrifugation and purified on Percoll gradient. In order to identify the viral particles used in the immunization procedure, the protein pattern of the particles was determined by electrophoresis and compared with the pattern of other known coronaviruses. Analysis of structural polypeptides of the purified viral particles revealed a pattern similar to that reported for other coronaviruses. These polypeptides cross reacted with two other coronavirus specific immune sera (IBV and TGE). IEM assay of fecal samples collected from healthy and sick rabbits showed the presence of immune aggregates in specimens from both sick and healthy rabbits. Those aggregates contained viral particles sharing morphological characteristics with other coronaviruses. Furthermore, IEM assay was shown to be more sensitive than a direct EM procedure to detect coronavirus particles in rabbit feces. This assay also allowed the detection of a larger number of chronic carriers.

Animals↗
PubMed1984

Morphology and ultrastructure of Lymphocystis disease virus, a fish iridovirus, grown in tissue culture.

The morphology and the ultrastructure of the Lymphocystis disease virus (LDV) strain Leetown , a fish iridovirus , was studied by electron microscopy. The virus was grown on bluegill fry (BF-2) cells at 21 degrees. LDV showed an icosahedral shape by ultrathin sections and negative staining, with a diameter of about 200 nm. The shell of the virion seemed to be composed of two unitary membranes with a total diameter of 16 nm. The outer membrane demonstrated swelling in negative staining, exhibiting a central core and the presence of globular subunits at its external surface organized in geometrical arrays of 60 and 90 degrees. Glutaraldehyde fixation preserved very effectively the icosahedral structure with the subunits remaining invisible. The internal structure of the virions was composed of osmiophilic threads or granules of 6 to 8 nm in diameter surrounded by an amorphous material of 10 to 20 nm in thickness. External filaments were observed at the surface of the particles in ultrathin sections, giving the appearance of a halo surrounding the shell. In negative staining these filaments were rarely observed; in one virus preparation, they appeared in bundles.

Animals↗
PubMed1982

Mammalian cells contain a DNA-recombining activity that can be dissociated from topoisomerase activity.

The role of the topoisomerase enzyme in DNA recombination was investigated by extracting chromosomal deoxyribonucleoproteins from a variety of cultured mammalian cells and assaying for the formation of recombinant DNA structures. Although each of the crude deoxyribonucleoprotein preparations contained topoisomerase activity, they did not all contain DNA-recombining activity. A distinct, perhaps novel, enzyme may therefore promote DNA recombination in these cell-free systems.

Animals↗
PubMed1982

Electron microscopic study of bluegill virus.

Bluegill virus (BGV) grown in BF-2 cells was studied by negative staining and ultrathin sections of infected cells. Although BGV resembles bunyaviruses in gross physical appearance, it differs from this group in several important aspects. Thus, BGV cannot be classified as a member of the Bunyaviridae family and could be a representative of a novel family of enveloped RNA viruses.

Animals↗
PubMed1981

Comparison of three electron microscopy techniques for the detection of human rotaviruses.

Rotavirus detection by direct electron microscopy was compared with direct and indirect immune electron microscopy techniques. The latter two approaches permitted the enumeration of 25 and 103 times more rotaviruses respectively, than direct electron microscopy. Also, 70% and 90% of the virus particles were aggregated by direct and indirect immune electron microscopy techniques respectively, thus facilitating their detection.

Antibodies, Viral↗