Search PubMed⌕ Search

Biomedical subjects

M D Parker

Publications and source records attributed to M D Parker.

At least 37 records · Page 2Linked to original sources
PubMed1991

Rotavirus particles function as immunological carriers for the delivery of peptides from infectious agents and endogenous proteins.

A major problem in the development of useful animal subunit vaccines has been the generation of immune responses to weakly immunogenic molecules. For this purpose a new and effective delivery system has been devised. This system is based upon the inner capsid of bovine rotavirus. Under the appropriate conditions, the inner capsid protein, designated BP6, can be made to self-assemble in vitro and form spherical particles. These particles possess an inherent capacity to target to cells of the immune system. Exploitation of these properties has led to the development of technology to couple antigens to the VP6 particles such that the sphere acts as a novel immunological carrier. This is based on a "binding peptide" derived from another rotavirus peptide, VP4, as well as on more traditional techniques of chemical coupling. We have coupled peptides or proteins to this carrier via the binding peptide and have shown that every epitope tested to date gave excellent immune responses. Furthermore, using this carrier, immunity has been developed without the use of adjuvants. This has far-reaching implications for animal and human immunization.

Amino Acid Sequence↗
PubMed1991

Expression of bovine herpesvirus 1 glycoprotein gIV by recombinant baculovirus and analysis of its immunogenic properties.

The gene encoding the gIV glycoprotein of bovine herpesvirus 1 has been inserted into the genome of Autographa californica baculovirus in lieu of the coding region of the A. californica baculovirus polyhedrin gene. Recombinant protein was identified by its reactivity with gIV-specific monoclonal antibodies and expressed at high levels (about 85 micrograms per 2.5 x 10(6) cells) in Spodoptera frugiperda (SF9) cells. The recombinant glycoprotein had an apparent molecular mass of 63 kDa, indicating that it was incompletely glycosylated. However, it was transported to and expressed on the cell surface of infected SF9 cells. Furthermore, reactivity with polyclonal and monoclonal antibodies specific for gIV suggested that most epitopes were functionally unaltered on the recombinant gIV. Immunization of cattle with recombinant gIV in crude, partially purified, or pure form resulted in the induction of neutralizing antibodies to BHV-1, which were reactive with authentic gIV. However, the neutralizing antibody titers were lower than those elicited by an equivalent amount of affinity-purified authentic gIV, which appeared to be mainly due to reduced recognition of one of the neutralizing antigenic domains of gIV, designated domain I. The potential use of this recombinant gIV glycoprotein as a vaccine to bovine herpesvirus 1 infection in cattle is discussed.

Animals↗
PubMed1991

Heterotypic passive protection induced by synthetic peptides corresponding to VP7 and VP4 of bovine rotavirus.

We have evaluated the potential of two peptides derived from highly conserved regions of rotavirus outer capsid proteins (VP7 and VP4) to act as a rotavirus vaccine. The capacity of peptides coupled to rotavirus VP6 spherical particles to provide passive protection in a murine model was compared with the protection induced by peptide-keyhole limpet hemocyanin (KLH) conjugates. Female mice were immunized a total of three times before and during pregnancy. Suckling mouse pups were challenged at 7 days of age with either homologous or heterologous rotavirus serotypes. The efficacy of vaccination was determined by analyzing the clinical symptoms and measuring xylose adsorption in the intestine. In this model the VP4 peptide-VP6 conjugate provided protection equal to that obtained using bovine rotavirus (BRV) as the immunogen. The VP7 peptide-VP6 conjugate provided slightly less protection than the VP4 peptide-VP6 conjugate. A mixture of the VP4 peptide-VP6 and VP7 peptide-VP6 conjugates provided better heterologous protection than immunization with BRV. In contrast, KLH-conjugated peptides provided only partial protection. The significance of a synthetic-peptide-based rotavirus vaccine in the prevention of rotavirus infections is discussed.

Amino Acid Sequence↗
PubMed1990

Analysis of the S spike (peplomer) glycoprotein of bovine coronavirus synthesized in insect cells.

The bovine coronavirus (BCV) spike glycoprotein precursor (S, formerly termed peplomer) and its two subunit polypeptides (S1 and S2) were individually expressed in Spodoptera frugiperda (Sf9) insect cells. Each recombinant baculovirus expressed both glycosylated (S, 170K; S1, 95K; S2, 80K) and unglycosylated (S0, 140K; S10, 75K; and S20, 65K) forms of BCV spike polypeptides in Sf9 cells. The mature 95K S1 polypeptide was secreted whereas the S and S2 polypeptides remained cell-associated. The S precursor was partially cleaved in Sf9 cells, and the resulting S1 was also released into the medium. Neutralizing monoclonal antibodies representing two antigenic domains bound to recombinant S and S1 but not the S2 polypeptides, indicating that two major epitopes for BCV neutralization are located on the S1 subunit.

Amino Acid Sequence↗
PubMed1990

Primary structure of the S peplomer gene of bovine coronavirus and surface expression in insect cells.

The nucleotide sequence of the S peplomer gene of bovine coronavirus (BCV) has been determined. A single open reading frame of 4089 nucleotides encodes a polypeptide of 150K with 20 potential sites for addition of N-linked oligosaccharides. Expression of the cloned BCV S gene by a recombinant of Autographa californica nuclear polyhedrosis virus resulted in production of a 180K glycosylated polypeptide which was transported to the surface of the cell. Comparison of the BCV S gene with the analogous genes of murine hepatitis viruses shows that the BCV S polypeptide contains a unique domain of 138 amino acids not present in murine hepatitis virus strain JHM, but which has a partially homologous counterpart in strain A59. This domain accounts for most of the differences in size of the S gene products of these coronaviruses.

Amino Acid Sequence↗
PubMed1990

Expression and secretion of the bovine coronavirus hemagglutinin-esterase glycoprotein by insect cells infected with recombinant baculoviruses.

A cDNA fragment representing the hemagglutinin-esterase (HE) gene of bovine coronavirus (BCV) was inserted into the genome of Autographa californica nuclear polyhedrosis virus. Infection of insect cells with the recombinant virus resulted in the production of a 120-kilodalton disulfide-linked dimeric form of the BCV HE polypeptide. Deletion of the carboxy-terminal hydrophobic domain from the HE polypeptide resulted in secretion of a dimeric form of the truncated HE polypeptide. The acetylesterase activity of the BCV HE was detectable in insect cells expressing the BCV hemagglutinin and was inhibited by two monoclonal antibodies which also inhibit hemagglutination.

Acetylesterase↗
PubMed1989

Dysphagia due to cervical osteophytes: a controversial entity revisited.

Although cervical osteophytes have been reported to cause dysphagia, most remain clinically silent. A case of osteophytic dysphagia is reported in which an unusually large osteophytic ridge caused marked deviation and narrowing of the cervical esophagus. The subject of osteophytic dysphagia is reviewed.

Aged↗
PubMed1989

Evolving concepts in the diagnosis of xanthogranulomatous pyelonephritis.

Xanthogranulomatous pyelonephritis (XGP) is a rare inflammatory process of the kidney that results in focal or diffuse renal enlargement and nonexcretion. In the past, XGP often has been misdiagnosed as renal carcinoma. Newer investigative modalities and an increased awareness of XGP should make preoperative diagnosis possible. Thirty-two cases of XGP are presented, with emphasis on recent advances in radiographic approach to diagnosis.

Adult↗
PubMed1989

Multiple ureteral diverticula: a possible radiographically demonstrable risk factor in development of transitional cell carcinoma.

Six cases of multiple ureteral diverticula are reported. Most were associated with urinary tract obstruction or stasis and infection. One-third had transitional cell carcinoma, a frequency similar to that reported in previous series. These diverticula are a radiographically demonstrable reflection of urothelial hyperplasia/metaplasia and may represent a risk factor for development of transitional cell carcinoma.

Adult↗
PubMed1989

Baculovirus expression of the M genome segment of Rift Valley fever virus and examination of antigenic and immunogenic properties of the expressed proteins.

Autographa californica nuclear polyhedrosis viral recombinants containing coding information for the Rift Valley fever virus (RVFV) envelope glycoproteins (G1 and G2) and varying amounts of preglycoprotein coding sequences were prepared by using transfer vectors pAc373 or pAcYM1. Expression products were processed to yield proteins indistinguishable from authentic G1 and G2 by gel electrophoresis. The immunogenic properties of the expressed proteins were assessed by immunizing mice and challenging with RVFV. A single inoculation with lysates of cells infected with recombinants expressing both G1 and G2 induced neutralizing antibody responses in mice and protected them from an otherwise lethal challenge with RVFV. Lysates of cells infected with a recombinant expressing only G2 also induced a protective response after two immunizations. Survivors displayed elevated antibody titers to G1 and G2 and also developed antibodies to the RVFV nucleocapsid protein, the latter allowing discrimination from vaccinated mice and indicating that animals had survived infection. Nonimmune mice were protected from lethal RVFV infection by passive transfer of sera from animals immunized with recombinant antigens, indicating that a humoral immune response is sufficient to protect against RVFV.

Animals↗
PubMed1989

Streptococcal antigen-induced dislocation and dysplasia of the hip in newborn rats. Radiologic and histologic evaluation of a model of congenital dislocation of the hip.

Dislocation of the hip developed in 62% of newborn rats with streptococcal antigen-induced synovitis. Age at the time of the induction of synovitis is critical since dislocation is not observed in older rats. Synovitis with distention and laxity of the joint capsule is most likely responsible for the hip dislocation. Although congenital dislocation of the hip in children is not mediated by an inflammatory process, the current model of dislocation of the hip in rats is similar in being critically age-dependent, and associated with ligamentous laxity. Our model may be helpful in studying this important clinical entity.

Age Factors↗
PubMed1989

Cloning and in vitro expression of the gene for the E3 haemagglutinin glycoprotein of bovine coronavirus.

A cDNA clone representing the gene for the E3 glycoprotein, the haemagglutinin, of bovine coronavirus was isolated from a plasmid cDNA library of the viral genome and sequenced. The gene is located immediately 5' of the E2 glycoprotein gene on the viral genome. Nucleotide sequencing of the E3 gene predicts a polypeptide of 424 amino acids with an Mr of 47K. In vitro translation of mRNA transcribed from the cloned E3 gene yielded a polypeptide of Mr 45K, similar to that predicted from the nucleotide sequence. In the presence of microsomal membranes, the in vitro product was cotranslationally processed to a 62K polypeptide which comigrated on SDS-polyacrylamide gels with the E3 monomer (gp62) obtained from virus-infected cells. Both the 45K and 62K polypeptides were immunoprecipitated with E3-specific monoclonal antibodies, confirming the identity of the gene as that encoding the E3 glycoprotein. Finally, only monoclonal antibodies to the E3 protein inhibited haemagglutination by the virus thus confirming its identity as the haemagglutinin of bovine coronavirus.

Amino Acid Sequence↗
PubMed1989

Mapping of neutralizing epitopes to fragments of the bovine coronavirus E2 protein by proteolysis of antigen-antibody complexes.

Neutralizing antigenic domains on bovine coronavirus gp100/E2 were mapped to fragments of this protein by proteolytic cleavage and fragment analysis. The procedure involved analysis of fragments generated after incubation of E2-monoclonal antibody complexes with various proteases. The smallest antibody-bound fragments obtained were a 50K fragment following Staphylococcus aureus V8 protease and submaxillary protease digestion, and a 37K fragment following trypsin digestion. Trypsin also produced a transient antibody-bound 50K fragment. A 40K fragment which was not bound by antibody was observed following digestions with all three proteases. The 50K fragments generated by V8, submaxillary protease and trypsin comigrated on gels and displayed the same altered mobility under non-reducing conditions, suggesting identity of these fragments and indicating the presence of disulphide linkages in these fragments. The 40K fragments generated by these three enzymes also comigrated and displayed the same altered mobility under non-reducing conditions. The 37K trypsin fragment contained both neutralizing domains, A and B.

Animals↗
PubMed1989

Disappearing breast calcifications.

Seventeen patients aged 47-67 years had 20 foci of disappearing breast calcifications. Firm compression was used to obtain craniocaudal and oblique mediolateral views of each patient. The authors categorized calcifications according to shape, size, number, position, and likelihood of benignity or malignancy. The right breast was the site of disappearing calcifications more often than the left. The most common configuration of disappearing calcifications was round or oval. All calcifications but one were found in dense glandular areas of the breast. Dense glandular tissue within 1 cm of the border with the stroma was the most common location. Only one of 20 foci suggested malignancy. The disappearance of breast calcifications is uncommon but probably not rare.

Aged↗