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M J Hamilton

Publications and source records attributed to M J Hamilton.

At least 37 records · Page 2Linked to original sources

Induction of type 2 cytokines by a staphylococcal enterotoxin superantigen.

Persistent intramammary infections of dairy cows with Staphylococcus aureus may involve immunosuppression mediated by bacterial toxins such as enterotoxins and other super-antigens (SAgs). Previously we found that stimulation of bovine PBMC with staphylococcal enterotoxin C (SEC) induced a unique phenotype of activated CD8+ T cells expressing a newly identified activation molecule, ACT3. In the present study we found that SEC induced the expression of interleukin (IL)-4 and IL-10 mRNAs, two cytokines associated with type 2 responses. Elevated levels of IL-4 and IL-10, observed between day 0 and day 4 of culture, were associated with temporary inhibition of proliferative responses of T cells, evidenced by a decrease in numbers of CD4+ T cells and a small increase in numbers of CD8+ T cells. Vigorous proliferation of T cells occurred between days 4 and 7 of culture and with a bias towards CD8+ T cells. Acquisition of the ACT3+ phenotype by CD8+ T cells was preceded by induction of IL-4 mRNA. Thus, in the bovine system, SAgs may hinder protective responses by inducing type 2 cytokines, which interfere with immune clearance of many microbial pathogens. The results of the study are consistent with the hypothesis that SAgs are involved in immunosuppression, and suggest possible immunomodulatory mechanisms.

Animals↗

DNA synapomorphies for a variety of taxonomic levels from a cosmid library from the New World bat Macrotus waterhousii.

An effective method yielding taxon-specific markers from the genome of a single individual would be valuable for many types of scientific investigations, including systematic, forensic, conservation, and evolutionary studies. We explored the use of cosmid libraries, with insert sizes averaging 35 kb, to streamline the process of locating sequences of DNA that can serve as taxonomic markers from the specific to the ordinal levels. By screening approximately 2.6% of the leaf-nosed bat (Macrotus waterhousii) genome, we identified several potential DNA fragments that appear to be synapomorphic for a variety of taxonomic levels. A more thorough analysis of the markers documented that 17 Macrotus-specific clones represent three distinct DNA generic markers, whereas 30 microchiropteran clones represent multiple copies of a single family of repetitive DNA. The Microchiroptera taxon markers hybridize with representatives of most of the Microchiroptera families; however, no hybridization was detected for members of the superfamily Rhinolophoidea. These results demonstrate that cosmid libraries can be a valuable source for isolating taxon-specific markers from mammals even when the insert size is as large as 35 kb.

Animals↗

High levels of genetic change in rodents of Chernobyl.

Base-pair substitution rates for the mitochondrial cytochrome beta gene of free-living, native populations of voles collected next to reactor 4 at Chernobyl, Ukraine, were estimated by two independent methods to be in excess of 10(-4) nucleotides per site per generation. These estimates are hundreds of times greater than those typically found in mitochondria of vertebrates, suggesting that the environment resulting from this nuclear power plant disaster is having a measurable genetic impact on the organisms of that region. Despite these DNA changes, vole populations thrive and reproduce in the radioactive regions around the Chernobyl reactor.

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Analysis of monoclonal antibodies specific for the gamma delta TcR.

gamma delta T cells in ruminants can be subdivided in two or more subpopulations on the basis of the expression of surface antigen WC1, which can exist in different isoforms. In this study, 18 monoclonal antibodies (mAbs) submitted to the Third International Workshop that were predicted to react with gamma delta TcR molecules were analysed and expression of their antigens was investigated on the different gamma delta T cell subpopulations. A set of control mAbs positive for TcR1 (86D), BoCD3 (MM1A), WC1 (B7A1, BAQ4A, CACTB32A, and BAQ89A) was included for comparative studies. Previous investigations demonstrated eight of the mAbs immunoprecipitated peptides with apparent M(r)s of 37 and/or 47 kDa, indicating they recognized determinants on the T cell receptor, TcR1. Two color flow cytometric analyses in the present study demonstrated the mAbs formed three groups; group 1, a set of mAbs that recognize TcR1 determinants expressed on all gamma delta T cells and groups 2 and 3, sets of mAbs that recognize TcR1 determinants on some gamma delta T cells: TcR1-N6 and TcR1-N7 respectively. mAbs from the latter groups define families of TcR1 molecules that express either one or both of the determinants. These antigenically distinct forms of TcR1 are expressed in equal proportion on the two gamma delta T cell populations that express one of the mutually exclusive isoforms of WC1, WC1-N3 and WC1-N4. The data indicate usage of the mAb-defined families of the gamma delta TcR is primarily restricted to the WC1+ subpopulation of gamma delta T cells. However, a small subpopulation of CD2+, WC1- gamma delta T cells expresses a form of TcR1 positive for the determinant TcR1-N6.

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Analysis of monoclonal antibodies reactive with molecules upregulated or expressed only on activated lymphocytes.

Monoclonal antibodies potentially specific for antigens expressed or upregulated on activated leukocytes were selected for further analysis from the panel submitted to the third international workshop on ruminant leukocyte antigens. The kinetics of expression of these activation antigens on resting peripheral mononuclear cells (PBMC) and PBMC stimulated with concanavalin A or staphylococcal superantigen SECI for 4, 24 or 96 h were compared, as well as their appearance on various subsets of cells. For some of them, a molecular mass could be determined after immunoprecipitation from radio-labeled, lectin-stimulated cells. Based on the results from the clustering, kinetic studies and biochemical data, evidence was gathered for assigning two additional mAbs to cluster BoCD25 (IL-2 receptor) and two mAbs to cluster BoCD71 (transferrin receptor). Four mAbs recognized an early activation antigen predominantly expressed on gamma delta T cells in short-term cultures. A number of other activation antigens were further characterized.

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Early speech changes in children with multichannel cochlear implants.

A number of studies have demonstrated that cochlear implants provide an improved auditory signal and enhance the development of speech-perception and production skills for profoundly deaf children. However, exactly when these early speech skills begin to develop remains unclear. To explore this issue, we observed, for a 1-year period, four prelingually deaf children who underwent implantation consecutively within 1 month of each other, and we paid particular attention to the first few months of rehabilitation. We found immediate speech scores as early as the first day of implant tune-up. Speech production continued to improve rapidly throughout the first 4 months but exhibited a generally slower rate of progress in some of the speech-production skills at 1 year. We also found vowel-production skills to be the easiest to achieve, with word-pattern recognition and consonant voicing of intermediate difficulty. Consonant placing and manner of consonant production were the hardest skills to achieve. Results of speech-perception tests 1 year after implantation were markedly improved over preimplantation levels in three of the four children. These early speech changes stress the need for maximization of the capability of the cochlear implant by institution of immediate and intensive speech rehabilitation efforts for prelingually deaf children.

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Platelet deposition and fibrinogen binding on surfaces coated with heparin or friction-reducing polymers.

The blood-contacting properties of polyethylene coated with a lubricious hydrophilic coating; an uncoated polyethylene; or a photoheparin-treated polyethylene-negative control sample were compared by measuring fibrinogen adsorption, antifibrinogen binding, and platelet attachment from human plasma. The polyethylene surfaces coated with a hydrophilic polymer were found to be similar to surfaces coated with heparin. Fibrinogen adsorption on the hydrophilic coating was 60% lower than adsorption on either the uncoated or heparin-coated polyethylene samples. Antifibrinogen binding from buffer to the hydrophilic coating was also reduced more than 85% from binding to uncoated polyethylene samples. Both the hydrophilic coating and heparin coating showed a reduction in platelet attachment by a factor of 100 over the uncoated sample as well as significantly reduced platelet activation.

Acrylic Resins↗

Differential distribution of gamma delta T-cell receptor lymphocyte subpopulations in blood and spleen of young and adult cattle.

A panel of monoclonal antibodies to bovine leukocyte differentiation molecules was used to evaluate peripheral blood and splenic lymphocytes from cattle of various ages. The major population of peripheral blood lymphocytes from neonatal calves was gamma delta T-cell receptor (TCR1) positive, as determined by TCR1-N12 expression. TCR1-N12+ lymphocytes were decreased in number in older calves, and were lowest in adult cattle. The major subpopulation of TCR1-N12+ cells from peripheral blood coexpressed WC1, but not BoCD2. A small subpopulation of peripheral blood TCR1-N12+ cells from cattle of all ages coexpressed BoCD2, but not WC1. The TCR1-N12+ BoCD2+ lymphocytes made up the largest TCR1-N12+ lymphocyte subpopulation in spleens of both calves and adults. The TCR1-N12+WC1+ splenic lymphocytes were present as a small population. The data indicate that two subpopulations of TCR1+ lymphocytes are present in cattle of all ages. These two subpopulations are differentially distributed between blood and spleen, with TCR1-N12+WC1+ lymphocytes predominating in blood, and TCR1-N12+BoCD2+ cells predominating in spleen.

Aging↗

Cytogenetic nomenclature of deer mice, Peromyscus (Rodentia): revision and review of the standardized karyotype. Report of the Committee for the Standardization of Chromosomes of Peromyscus.

A revision of the standardized karyotype of deer mice (Peromyscus) is presented. This revision addresses short-comings of the original standardization, contains a substantial increase in the number of G-band markers and provides a nomenclature for the G-bands of each autosome and the X chromosome. Using the revised standardized karyotype, we specify the particular G-bands or patterns that identify each chromosome and catalog the more problematic chromosome identifications and likely misidentifications. For each chromosome, we present an overview of previously reported variation in euchromatic arrangement and heterochromatic constitution. We then review previous applications of the standardized karyotype and summarize the predominant findings from cytogenetic and cytosystematic studies of Peromyscus and related taxa.

Animals↗

Suppression of proliferative response of BoCD4+ T lymphocytes by activated BoCD8+ T lymphocytes in the mammary gland of cows with Staphylococcus aureus mastitis.

Investigations were conducted to determine the mechanisms that account for differences in the responses of BoCD4+ lymphocytes from mammary gland secretions (MGS) in healthy cows and in cows with Staphylococcus aureus infection. The proliferative response to lectins and S. aureus antigens of mammary gland lymphocytes from healthy, S. aureus immunized cows was less than the response of peripheral blood lymphocytes. The lower responses of mammary gland lymphocytes were attributable both to less efficient antigen presentation by mammary gland antigen-presenting cells (APC) than by peripheral blood APC, and to lower responsiveness of mammary gland lymphocytes to lectins and antigen. In addition, the proliferative response of infected mammary gland lymphocytes was less than the response of uninfected mammary gland lymphocytes. This difference resulted from decreased proliferation of BoCD4+ lymphocytes in infected MGS. Flow cytometric analysis revealed that infected MGS contained increased numbers of BoCD8+ cells which coexpressed an activation molecule, ACT2, relative to BoCD8+ cells from uninfected MGS. Removal of BoCD8+, ACT2+ lymphocytes resulted in increased antigen responsiveness by lymphocytes from infected mammary glands. Also, when purified BoCD4+ lymphocytes were stimulated with antigen in the presence of varying numbers of ACT2+, BoCD8+ lymphocytes, antigen responsiveness was decreased in a dose-related manner. These data demonstrate that hyporesponsiveness of mammary gland lymphocytes to lectins and S. aureus antigen is, in part, mediated by activated BoCD8+ lymphocytes and suggest that this population enhances persistent intramammary infection by S. aureus.

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Identification of a monoclonal antibody reactive with the bovine orthologue of CD3 (BoCD3).

A monoclonal antibody (mAb), MM1A, that identifies a molecule expressed on a large percentage of bovine lymphocytes (60-80%) was examined to determine its specificity. Two and three colour immunofluorescence analysis using flow cytometry revealed the molecule is highly expressed on all CD4+ and CD8+ lymphocytes and lymphocytes that express WC1 and the gamma/delta TCR. In contrast to mAbs reactive with BoCD5, MM1A did not react with B lymphocytes. Biochemical analysis revealed that the mAb immunoprecipitates a molecular complex comprising a set of peptides with M(r) approximately 12, 16, 22, 32, 36, and 44 kDa under reducing conditions and an additional 96 kDa peptide complex under non-reducing conditions. The data indicate that MM1A recognizes the bovine orthologue of CD3 (BoCD3).

Animals↗

Molecular phylogenetics of Stenodermatini bat genera: congruence of data from nuclear and mitochondrial DNA.

Within the tribe Stenodermatini the systematics of the complex of species allied with the genus Artibeus has generated several alternative phylogenetic hypotheses. The most recent treatment recognized four genera (Artibeus, Dermanura, Enchisthenes, and Koopmania) and suggested that the most recent common ancestor of these four genera would include the common ancestor of all other currently recognized Stenodermatini genera except Sturnira. To test this hypothesis, we examined an EcoRI-defined nuclear satellite DNA repeat and 402 bp of DNA sequence variation from the mitochondrial cytochrome b gene. Phylogenetic conclusions based on Southern blot analyses, in situ hybridization, and mitochondrial DNA sequence data indicate that Enchisthenes is not closely related to Dermanura, Artibeus, or Koopmania and that Dermanura, Artibeus, and Koopmania shared a common ancestor after diverging from the remainder of the Stenodermatini. If our conclusions are correct, then justification for recognizing Dermanura and Koopmania as generically distinct from Artibeus must be based on the magnitude of difference that distinguishes each rather than on the conclusion that to place them as congeneric with Artibeus creates a paraphyletic taxon.

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Transposable elements and the evolution of genome organization in mammals.

All mammalian transposable elements characterized to date appear to be nonrandomly distributed in the mammalian genome. While no element has been found to be exclusively restricted in its chromosomal location, LINE elements and some retrovirus-like elements are preferentially accumulated in G-banding regions of the chromosomes, and in some cases in the sex chromosomes, while SINE elements occur preferentially in R-banding regions. Four mechanisms are presented which may explain the nonrandom genomic distribution of mammalian transposons: i) sequence-specific insertion, ii) S-phase insertion, iii) ectopic excision, and iv) recombinational editing. Some of the available data are consistent with each of these four models, but no single model is sufficient to explain all of the existing data.

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Intragenomic movement and concerted evolution of satellite DNA in Peromyscus: evidence from in situ hybridization.

Four DNA probes isolated from Peromyscus leucopus were used to examine intra- and interspecific variation in the chromosomal location of satellite DNA in the genus Peromyscus. All four probes hybridized to the centromeric regions of all chromosomes in all species of Peromyscus examined but did not hybridize to the majority of heterochromatic regions in closely related non-Peromyscus species. One probe contains a nonsatellite repetitive sequence. The implications of these data to the evolution of genome organization are discussed.

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A rapid, reliable method of evaluating growth and viability of intraerythrocytic protozoan hemoparasites using fluorescence flow cytometry.

Fluorescence flow cytometry was employed to assess the potential of a vital dye, hydroethidine, for use in the detection and monitoring of the viability of hemoparasites in infected erythrocytes, using Babesia bovis as a model parasite. The studies demonstrated that hydroethidine is taken up by B. bovis and metabolically converted to the DNA binding fluorochrome, ethidium. Following uptake of the dye, erythrocytes containing viable parasites were readily distinguished and quantitated. Timed studies with the parasiticidal drug, Ganaseg, showed that it is possible to use the fluorochrome assay to monitor the effects of the drug on the rate of replication and viability of B. bovis in culture. The assay provides a rapid method for evaluation of the in vitro effect of drugs on hemoparasites and for analysis of the effect of various components of the immune response, such as lymphokines, monocyte products, antibodies, and effector cells (T, NK, LAK, ADCC) on the growth and viability of intraerythrocytic parasites.

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