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Biomedical subjects

R Bishop

Publications and source records attributed to R Bishop.

At least 55 records · Page 3Linked to original sources

The role of Gulf Coast oysters harvested in warmer months in Vibrio vulnificus infections in the United States, 1988-1996. Vibrio Working Group.

Vibrio vulnificus infections are highly lethal and associated with consumption of raw shellfish and exposure of wounds to seawater. V. vulnificus infections were reported to the Centers for Disease Control and Prevention from 23 states. For primary septicemia infections, oyster trace-backs were performed and water temperature data obtained at harvesting sites. Between 1988 and 1996, 422 infections were reported; 45% were wound infections, 43% primary septicemia, 5% gastroenteritis, and 7% from undetermined exposure. Eighty-six percent of patients were male, and 96% with primary septicemia consumed raw oysters. Sixty-one percent with primary septicemia died; underlying liver disease was associated with fatal outcome. All trace-backs with complete information implicated oysters harvested in the Gulf of Mexico; 89% were harvested in water >22 degrees C, the mean annual temperature at the harvesting sites (P < .0001). Control measures should focus on the increased risk from oysters harvested from the Gulf of Mexico during warm months as well as education about host susceptibility factors.

Adolescent↗

Production and characterization of a new monoclonal antibody effective in recognizing the CD3 T-cell associated antigen in formalin-fixed embedded tissue.

Phenotypic analysis of lymphoproliferative disorders is now considered mandatory for accurate classification which is the basis for optimum patient management. This is presently carried out in most cases using a range of antibodies recognizing B and T-cell antigens effective in paraffin sections, and an antibody to CD 3 is currently a key member of such panels, indicating T-cell phenotype. Current antibodies to CD3 are polyclonal with the inherent disadvantages of this type of reagent compared to monoclonal antibodies. In this study, we have used a recombinant fusion protein representing part of the epsilon subunit of the CD3 molecule to generate a novel monoclonal antibody (NCL-CD3-PS1) effective in paraffin sections. The antibody has been characterized biochemically and by immunohistochemistry using a wide range of normal and pathological tissues. Lineage and phenotype specificity have been supported in our study and results from other laboratories are awaited with interest.

Antibodies, Monoclonal↗

Effects of creatine supplementation on repetitive sprint performance and body composition in competitive swimmers.

In a double-blind and randomized manner, 18 male and female junior competitive swimmers supplemented their diets with 21 g.day-1 of creatine monohydrate (Cr) or a maltodextrin placebo (P) for 9 days during training. Prior to and following supplementation, subjects performed three 100-m freestyle sprint swims (long course) with 60 s rest/recovery between heats. In addition, subjects performed three 20-s arm ergometer maximal-effort sprint tests in the prone position with 60 s rest/recovery between sprint tests. Significant differences were observed among swim times, with Cr subjects swimming significantly faster than P subjects following supplementation in Heat 1 and significantly decreasing swim time in the second 100-m sprint. There was also some evidence that cumulative time to perform the three 100-m swims was decreased in the Cr group. Results indicate that 9 days of Cr supplementation during swim training may provide some ergogenic value to competitive junior swimmers during repetitive sprint performance.

Adolescent↗

Concerted evolution at a multicopy locus in the protozoan parasite Theileria parva: extreme divergence of potential protein-coding sequences.

Concerted evolution of multicopy gene families in vertebrates is recognized as an important force in the generation of biological novelty but has not been documented for the multicopy genes of protozoa. A multicopy locus, Tpr, which consists of tandemly arrayed open reading frames (ORFs) containing several repeated elements has been described for Theileria parva. Herein we show that probes derived from the 5'/N-terminal ends of ORFs in the genomic DNAs of T. parva Uganda (1,108 codons) and Boleni (699 codons) hybridized with multicopy sequences in homologous DNA but did not detect similar sequences in the DNA of 14 heterologous T. parva stocks and clones. The probe sequences were, however, protein coding according to predictive algorithms and codon usage. The 3'/C-terminal ends of the Uganda and Boleni ORFs exhibited 75% similarity and identity, respectively, to the previously identified Tpr1 and Tpr2 repetitive elements of T. parva Muguga. Tpr1-homologous sequences were detected in two additional species of Theileria. Eight different Tpr1-homologous transcripts were present in piroplasm mRNA from a single T. parva Muguga-infected animal. The Tpr1 and Tpr2 amino acid sequences contained six predicted membrane-associated segments. The ratio of synonymous to nonsynonymous substitutions indicates that Tpr1 evolves like protein-encoding DNA. The previously determined nucleotide sequence of the gene encoding the p67 antigen is completely identical in T. parva Muguga, Boleni, and Uganda, including the third base in codons. The data suggest that concerted evolution can lead to the radical divergence of coding sequences and that this can be a mechanism for the generation of novel genes.

Amino Acid Sequence↗

Theileria: improved species discrimination using oligonucleotides derived from large subunit ribosomal RNA sequences.

Partial nucleotide sequences of large subunit ribosomal RNA (LSU rRNA) coding genes of Theileria parva and T. taurotragi were determined. Alignment of the T. parva and T. taurotragi sequences revealed species-specific regions within the LSU rRNA genes. A major species-variable region, located between the first and second conserved secondary structure domains of LSU rRNA, was identified. The sequence of the major species-variable region was determined for T. annulata, T. buffeli, and an unclassified Theileria species isolated from buffalo. Species-specific oligonucleotides complementary to LSU rRNA sequences were designed for five species of Theileria. The oligonucleotide probes discriminated between Theileria species when hybridized to amplified parasite LSU rRNA genes or directly to parasite rRNA. Three species-specific oligonucleotides derived from nonoverlapping sequences were developed for each of T. parva and T. taurotragi. A simplified procedure was developed in which T. parva- and T. taurotragi-specific oligonucleotides were used to discriminate the two species under nonstringent hybridization and washing conditions. Use of the nonstringent conditions also increased the sensitivity of target detection. Comparison of Theileria species-specific oligonucleotides derived from large subunit and small subunit rRNA genes demonstrated that, for closely related species, probes derived from LSU rRNA sequences had improved specificity.

Animals↗

Structure and sequence variation of the genes encoding the polymorphic, immunodominant molecule (PIM), an antigen of Theileria parva recognized by inhibitory monoclonal antibodies.

The polymorphic, immunodominant molecule (PIM) of Theileria parva is the predominant antigen recognized by sera from infected cattle and by monoclonal antibodies (mAb) used to differentiate parasite strains. As such, the antigen is under consideration as a diagnostic antigen, and since the mAbs can neutralize sporozoite infectivity in vitro, in immunization experiments. Initial comparison of two PIM cDNA sequences suggested that the PIM genes consist of conserved 5' and 3' termini flanking a central variable region. We present further evidence, based on sequence analysis, supporting this general structure for the PIM genes. Evidence is also presented for a single copy of the PIM gene per haploid genome, implying that the different versions of PIM are encoded by distinct alleles. The central variable region of the PIM allele from the T. parva (Marikebuni) stock was found to contain 13 copies of the tetrapeptide repeat Gln-Pro-Glu-Pro. We also detected point mutations in the 5' and 3' termini of the PIM alleles, including regions recognized by the neutralizing and typing mAb. This contrasted with the high sequence conservation of the two introns of the genes, suggesting that the protein is undergoing rapid evolution. Sequence comparison of PIM genes from buffalo- and cattle-derived parasites supported earlier results that the parasites infecting buffaloes constitute a more heterogeneous population than those from cattle.

Amino Acid Sequence↗

Simultaneous administration of oral rhesus-human reassortant tetravalent (RRV-TV) rotavirus vaccine and oral poliovirus vaccine (OPV) in Thai infants.

Rhesus-human reassortant tetravalent (RRV-TV) oral rotavirus vaccine was given at the same time as oral poliovirus vaccine (OPV) or inactivated parenteral poliovirus vaccine (IPV) to Thai infants at 2, 4 and 6 months of age. Sera for rotavirus antibody studies were taken prior to and one month after each vaccination. After the first dose of vaccine at 2 months of age, 37% of the infants receiving rotavirus vaccine with IPV but only 10% of those receiving it with OPV showed a seroconversion by rotavirus IgA ELISA antibody test (p < 0.001). Likewise, neutralizing antibody seroconversion rates in initially seronegative subjects to rhesus rotavirus type 3 (RRV-3) after the first dose of RRV-TV vaccine were higher if the vaccine was given with IPV (74%) than if given with OPV (39%) (p = 0.0069). After the second and third doses of vaccine, the rotavirus IgA ELISA and RRV-3-neutralizing antibody response rates were not different between groups. Development of neutralizing antibodies to human rotavirus serotypes 1, 2 and 4 in the first seven months of life in vaccinees receiving rotavirus vaccine with OPV tended to occur at a lower rate than in those receiving rotavirus vaccine with IPV but the antibody levels were not significantly different at 7 months of age. Poliovirus type 2 and type 3 antibody responses were not different in infants receiving the rotavirus vaccine with OPV as compared with infants receiving only OPV. The mean poliovirus type 1 antibody level was slightly but not significantly lower at 5 and 7 months of age in infants that received both rotavirus vaccine and OPV.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗

Vaccination of Thai infants with rhesus-human reassortant tetravalent oral rotavirus vaccine.

In a randomized double blind placebo-controlled study, the rhesus-human reassortant tetravalent oral rotavirus vaccine (dose 4 x 10(4) plaque-forming units) was evaluated in Thai infants immunized at ages 2, 4 and 6 months. To investigate dose responses and to compare vaccine-induced and naturally acquired rotavirus immunity in the study population blood specimens were collected before and 1 month after each vaccination and at 12 months of age. No adverse reactions attributable to the vaccine were detected in the vaccinees. Sixty-three of 94 (67%) vaccine recipients showed a seroconversion in rotavirus IgA enzyme-linked immunosorbent assay antibodies after one or more doses, whereas only 15 of 93 (16%) placebo-vaccinated control children showed an IgA enzyme-linked immunosorbent assay antibody response, suggestive of natural rotavirus infection, between 2 and 7 months of age. By measuring rhesus rotavirus-neutralizing antibodies a seroconversion was detected in 49% of the vaccinees and 14% of the controls between 2 and 7 months of age. The geometric mean titers of neutralizing antibodies to human rotavirus serotypes 1, 2, 3 and 4 after the completion of vaccinations and at 12 months of age were higher in the vaccinees than in the controls. It is concluded that, even though maternally acquired rotavirus antibodies are commonly present, the rhesus-human reassortant tetravalent vaccine is immunogenic in many Thai infants ages 2 to 6 months. The immunogenicity of this vaccine is enhanced by multiple doses.

Administration, Oral↗

Theileria parva: detection of genomic polymorphisms by polymerase chain reaction amplification of DNA using arbitrary primers.

Genomic DNA fingerprints were generated from Theileria parva piroplasm DNA by polymerase chain reaction amplification using arbitrary decamers as primers. Four of 12 primers tested generated amplified fragments which were polymorphic between different T. parva stocks or clones. Hybridization of radiolabeled total amplification products to Southern blots of T. parva piroplasm and uninfected bovine DNA confirmed that none of the amplified fragments were derived from any contaminating bovine DNA which may have been present in small quantities. Polymorphic DNA fragments were reproducibly amplified from independent DNA preparations made from cloned parasites. Individual amplified DNA fragments were gel purified and shown to hybridize to size-polymorphic EcoRI fragments in T. parva genomic DNA. Hybridization of individual amplified fragments to total fingerprints, generated using the same arbitrary primer, demonstrated that a single primer detected at least three independent genomic polymorphisms in T. parva DNA.

Animals↗

Worldwide experience with the CR326F-derived inactivated hepatitis A virus vaccine in pediatric and adult populations: an overview.

The worldwide experience to date with VAQTA, a highly purified formalin-inactivated hepatitis A vaccine containing alum-adjuvant, is reviewed. No serious adverse experience related to vaccination has been reported. The vaccine has proven highly immunogenic, with seroconversion detectable after a single dose in 90-99% of children 2-16 years old, and of adults under 77 kg (170 lb) body weight. There is a trend toward lower one-dose seroconversion rates with increasing age and with weight > 77 kg. Early seroconversion in the latter groups may require two 25-unit doses given 2, 4 or 8 weeks apart, or a higher priming dose. Seroconversion induced by this vaccine has been shown to signify protection from clinical hepatitis A disease. The few vaccines whose titers have waned to borderline levels responded anamnestically to a booster, suggesting that the vaccine induces an immune memory response and should provide long-term protection.

Adolescent↗

Serum and salivary responses to oral tetravalent reassortant rotavirus vaccine in newborns.

Serum and salivary responses of 95 infants to either a standard (4 x 10(4) plaque-forming units (PFU), 47 neonates) or a high dose (4 x 10(5) PFU, 48 neonates) of tetravalent reassortant rhesus rotavirus vaccine (administered at 2 days and at 6 weeks of age) were evaluated in a double-blind clinical trial. Serum and salivary IgA antibodies to the rotavirus group A common antigen were determined by ELISA and radioimmunoassay (RIA). Serum neutralizing antibodies to rhesus rotavirus were determined by fluorescent focus reduction assay. No significant differences in responses to the high versus standard dose were noted in serum or saliva. Response was influenced by cord blood antibodies. All infants who were cord blood-negative for rhesus rotavirus neutralizing antibodies (nine who received the standard dose and 20 who received the higher dose) had serum responses, compared with 42-70% of those who were cord blood-positive. The serum response rate recorded for babies with cord blood neutralizing titres > 1000 was 44%. Infants being bottle fed had a higher serum response rate than did babies being breast fed exclusively. If serum and salivary responses were combined, the response rate reached 80% for bottle fed infants. Thus, determination of serum responses alone underestimates vaccine 'take' in infants, and more so in highly endemic areas than in areas subject only to sporadic outbreaks. However, determination of salivary responses in newborn breastfed infants may be inaccurate, due to possible persistence of antibodies derived from colostrum or breast milk.

Antibodies, Viral↗

Detection of a carrier state in Theileria parva-infected cattle by the polymerase chain reaction.

Two sets of oligonucleotide primers, one derived from a repetitive sequence and the other from the gene encoding a 67 kDa sporozoite antigen of Theileria parva, were used to amplify parasite DNA from the blood of T. parva-infected carrier cattle using the polymerase chain reaction (PCR). PCR amplification products were obtained from 15 carrier cattle infected with one of 4 different T. parva stocks. Successful amplifications were performed using DNA from 2 cattle infected with T. p. parva Pemba Mnarani, 10 cattle infected with T. p. parva Marikebuni, 2 cattle infected with T. p. bovis Boleni and 1 animal infected with T. p. lawrencei 7014. No amplification products were obtained from any of 7 cattle which had been infected with the T. p. parva Muguga stock. A synthetic oligonucleotide, which hybridized specifically to T. p. parva Marikebuni DNA among 6 T. parva stocks tested, was designed using sequence data from within the region of the T. parva genome amplified by the repetitive sequence primers. The oligonucleotide was used to probe PCR products and to increase the sensitivity and specificity of carrier animal detection. Southern blot analysis using a T. parva repetitive sequence probe demonstrated the existence of restriction fragment length polymorphisms between parasites isolated from T. p. parva Marikebuni-infected carrier cattle. The use of the PCR and other methods of carrier animal detection are discussed.

Amino Acid Sequence↗

Performance of the Reflotron in Massachusetts' Model System for Blood Cholesterol Screening Program.

The precision, accuracy, and durability of the Reflotron were evaluated by the Massachusetts Model Systems for Blood Cholesterol Screening Program. Screenings were conducted in diverse community settings over 16 months. Fingerstick samples from 10,428 individuals were tested. None of the four analyzers met the 1992 standards for precision, although two met the 1992 standards for accuracy. More than 40% of Reflotron values differed from the reference laboratory values by upwards of 5%. More than 16% of individuals were misclassified in terms of their risk category. All four instruments malfunctioned during the project.

Bias↗

Strain specificity of bovine Theileria parva-specific cytotoxic T cells is determined by the phenotype of the restricting class I MHC.

To determine whether the major histocompatibility complex (MHC) phenotype of cattle could affect the parasite strain specificity of immunity to Theileria parva by influencing the antigenic specificity of Theileria-specific cytotoxic T lymphocytes (CTL), we investigated the parasite strain specificity of Theileria-specific CTL clones derived from cattle of different class I MHC phenotypes. Thirty-one class I-restricted CTL clones were generated from four cattle immunized with the Muguga stock of T. parva. The MHC restriction and parasite strain specificities were determined for each clone utilizing as targets, parasitized cell lines of different MHC phenotypes and cloned cell lines containing different parasite strains. CTL clones restricted by the same MHC determinant had similar parasite strain specificities. On the other hand, clones restricted by different MHC determinants exhibited different parasite strain specificities. This was true whether the clones were generated from the same animal or from different cattle and tested on a target cell line expressing both MHC determinants. These results provide strong evidence that differences in the strain specificities of CTL derived from animals immunized with the same parasite stock, are determined by the class I MHC phenotype of the immunized animal.

Animals↗

Does autonomic neuropathy in diabetes cause hearing deficits?

This study examined the effect of autonomic neuropathy on hearing in diabetes mellitus. Twelve diabetics with autonomic neuropathy and eleven healthy controls were examined for hearing loss using standard audiological tests. Several subjects in both groups had some abnormal hearing tests, but the incidence was no greater in the diabetic group. No group trends were apparent. Diabetic autonomic neuropathy is unlikely to be associated with hearing deficits.

Acoustic Impedance Tests↗

Studies on attenuation of rotavirus. A comparison in piglets between virulent virus and its attenuated derivative.

The development of rotavirus vaccines against acute gastroenteritis for human infants has been accorded a very high priority. Several vaccine candidates all of which are live cultivated strains of animal origin have been tested in humans. However the nature of attenuation of these viruses for humans is unknown. In this study we have attenuated a pig rotavirus by 15 sequential passages in cell culture after which the virus no longer causes diarrhoea in piglets. The pathogenesis of infection of the attenuated rotavirus strain (AT/76 P15) in gnotobiotic piglets was compared with that of the virulent parent strain (AT/76). The pattern of virus replication in the small intestine was judged by histology, disaccharidase assay, immunoperoxidase labelling of gut sections using group A specific rotavirus antibody, and rotavirus antigen assay of gut contents. The parent strain caused variable but extensive infection that resulted in the complete destruction of mature small intestinal enterocytes and villous contraction within 3 days. Membrane bound digestive enzymes were lost, and profound watery diarrhoea and dehydration resulted in causing piglets to become moribund. In contrast attenuated virus appeared to propagate at a much slower pace. Fewer infected epithelial cells were detected at any one time. Destruction of enterocytes was never extensive enough to cause marked mucosal changes in histology. Membrane bound digestive enzymes remained near normal levels and there was little or no diarrhoea. Virus replication ceased after 6 days. It is concluded that attenuation of the porcine rotavirus strain studied was associated with its decreased ability to propagate in enterocytes after adaption to culture.

Animals↗

Sch 38519, a novel platelet aggregation inhibitor produced by a Thermomonospora sp. Taxonomy, fermentation, isolation, physico-chemical properties, structure and biological properties.

The complex containing a new platelet aggregation inhibitor, Sch 38519, was recovered from the fermentation filtrate of Thermomonospora sp. SCC 1793. A chemically defined medium was developed which favored the production of Sch 38519. The antibiotic was isolated from the fermentation filtrate by absorption on macroreticular resin and further purified by ion exchange chromatography and reverse phase HPLC. Sch 38519 is an isochromanequinone structurally related to medermycin, lactoquinomycin and granaticin. It inhibits thrombin-induced aggregation of human platelets with an IC50 of 68 micrograms/ml. Sch 38519 is also active against Gram-positive and Gram-negative bacteria.

Actinomycetales↗