Biomedical subjects

D J Miller

Publications and source records attributed to D J Miller.

At least 91 records · Page 5Linked to original sources

PubMed1997

Internal and external relationships of the Cnidaria: implications of primary and predicted secondary structure of the 5'-end of the 23S-like rDNA.

Since both internal (class-level) and external relationships of the Cnidaria remain unclear on the basis of analyses of 18S and (partial) 16S rDNA sequence data, we examined the informativeness of the 5'-end of the 23S-like rDNA. Here we describe analyses of both primary and predicted secondary structure data for this region from the ctenophore Bolinopsis sp., the placozoan Trichoplax adhaerens, the sponge Hymeniacidon heliophila, and representatives of all four cnidarian classes. Primary sequence analyses clearly resolved the Cnidaria from other lower Metazoa, supported sister group relationships between the Scyphozoa and Cubozoa and between the Ctenophora and the Placozoa, and confirmed the basal status of the Anthozoa within the Cnidaria. Additionally, in the ctenophore, placozoan and sponge, non-canonical base pairing is required to maintain the secondary structure of the B12 region, whereas amongst the Cnidaria this is not the case. Although the phylogenetic significance of this molecular character is unclear, our analyses do not support the close relationship between Cnidaria and Placozoa suggested by previous studies.

Animals↗

PubMed1997

Measuring psychological distress in candidates for liver transplantation: a pilot study.

A protocol to evaluate the psychopathology of candidates for orthotopic (whole organ) liver transplantation (OLTX) is presented. Patients being considered for OLTX at the VA Pittsburgh Healthcare System were routinely referred for psychological evaluation consisting of a clinical interview and battery of standardized tests. The findings on 26 patients corroborate previous reports of psychological distress among transplant candidates. There is also evidence that higher levels of disturbance are observed among patients with a positive history of substance abuse. This study supports the value of standardized testing in the transplant population and suggests directions for further research.

Adult↗

PubMed1997

Preliminary characterisation of an Onchocerca volvulus aspartic protease.

PCR using 1 primer specific for aspartic proteases and 1 primer annealing to the vector allowed amplification of a 377 bp fragment encoding part of an aspartic protease from an Onchocerca volvulus cDNA library. Use of this fragment as a probe allowed the isolation of a larger cDNA clone. In common with 2 other nematode aspartic proteases, the O. volvulus predicted protein has several of the general characteristics of this class of proteins, but lacks specific determinants of lysosomal localisation.

Amino Acid Sequence↗

PubMed1997

Successful treatment of established relapsing experimental autoimmune encephalomyelitis in mice with a monoclonal natural autoantibody.

We postulated that humoral autoimmunity can play a beneficial role in CNS demyelinating diseases such as multiple sclerosis. We previously demonstrated that monoclonal natural autoantibody SCH94.03 suppresses CNS inflammation and promotes remyelination in a virus-induced model of chronic progressive multiple sclerosis. To further investigate the relationship between natural autoimmunity and CNS demyelination, we examined the effect of SCH94.03 treatment on clinical relapses and pathological disease in SJL/J mice with established adoptive-transfer relapsing experimental autoimmune encephalomyelitis. Treatment with SCH94.03 after recovery from the initial episode of clinical disease reduced relapse rates by half, prolonged relapse onset by 6 days and reduced spinal cord demyelination and meningeal inflammation by 40%. These results are consistent with the hypothesized immunomodulatory function of natural autoantibodies, and are the first direct demonstration that natural humoral autoimmunity can be beneficial in an autoimmune T-cell-mediated CNS demyelinating disease.

Animals↗

PubMed1997

Variation in the ribosomal internal transcribed spacers and 5.8S rDNA among five species of Acropora (Cnidaria; Scleractinia): patterns of variation consistent with reticulate evolution.

The ITS sequences of Acropora spp. are the shortest so far identified in any metazoan and are among the shortest seen in eukaryotes; ITS1 was 70-80 bases, and ITS2 was 100-112 bases. The ITS sequences were also highly variable, but base composition and secondary structure prediction indicate that divergent sequence variants are unlikely to be pseudogenes. The pattern of variation was unusual in several other respects: (1) two distinct ITS2 types were detected in both A. hyacinthus and A. cytherea, species known to hybridize in vitro with high success rates, and a putative intermediate ITS2 form was also detected in A. cytherea; (2) A. valida was found to contain highly (29%) diverged ITS1 variants; and (3) A. longicyathus contained two distinct 5.8S rDNA types. These data are consistent with a reticulate evolutionary history for the genus Acropora.

Animals↗

PubMed1997

Sperm plasma membrane remodeling during spermiogenetic maturation in men: relationship among plasma membrane beta 1,4-galactosyltransferase, cytoplasmic creatine phosphokinase, and creatine phosphokinase isoform ratios.

Sperm creatine phosphokinase (CK) concentrations and the synthesis of the CK-M isoform reflect normal spermiogenesis and predict maturity and fertilizing potential of ejaculated human spermatozoa. Immature spermatozoa, characterized by cytoplasmic retention and low CK-M to CK-B isoform ratios, are deficient in zona binding and fail to cause pregnancies. Because these sperm lack zona-binding ability, we examined in this study whether beta 1,4-galactosyltransferase (GalTase), a key element of sperm-zona interactions in mice, is diminished in immature human sperm. Unexpectedly, GalTase was overexpressed in immature sperm relative to mature sperm: the levels of cytoplasmic CK and plasma membrane GalTase were positively correlated (r = 0.78, p < 0.001, n = 88). Sperm populations with various levels of cellular maturity, prepared by Percoll gradients, had different CK and GalTase concentrations, but within each subpopulation the relationship between CK and GalTase was maintained (p < 0.01-0.001). GalTase activities in intact and vortex-disrupted sperm fractions were similar, showing that GalTase is present on the surface membrane of human sperm--similar to the situation in all other species assayed. The changes previously reported by our laboratory in zona-binding ability and lipid peroxidation rates (which occur simultaneously with cytoplasmic extrusion), decline in CK activity, and increased expression of the CK-M isoform are suggestive of a remodeling of the sperm surface concomitant with cytoplasmic maturation. The changes reported here in GalTase expression on the surface of maturing spermatozoa prove this hypothesis.

Animals↗

PubMed1997

Sperm from a variety of mammalian species express beta1,4-galactosyltransferase on their surface.

In mice, initial gamete recognition is mediated by the binding of sperm surface beta1,4-galactosyltransferase (GalTase) to a glycoprotein of the zona pellucida, ZP3. When sperm bind to the zona pellucida, ZP3 induces the acrosome reaction by aggregating GalTase. The acrosome reaction releases acrosomal enzymes allowing sperm to pass through the zona pellucida, bind to the egg membrane, and activate development. In addition to GalTase, there is evidence that other sperm proteins may also bind ZP3. Although fertilization in the mouse is morphologically similar to fertilization in most other mammalian species, the degree of parallelism at the molecular level is not well defined. Less information is available about the molecular details of fertilization in other species. The aim of this work was to determine whether sperm from other mammalian species express GalTase on their surface. We performed GalTase enzyme assays on sperm from six species, and all six expressed GalTase on their surface. The amounts of GalTase varied between species. Guinea pig, mouse, and rat sperm had higher levels of GalTase than bovine, porcine, and rabbit sperm. GalTase was localized by immunofluorescence on live and fixed sperm to the anterior portion of the sperm head in all species examined. This is the expected location for a receptor that binds the zona pellucida. Biotinylation of sperm surface proteins confirmed that GalTase detected by immunofluorescence and enzyme assay was expressed on the sperm surface. These results demonstrate that various mammalian species express GalTase on their surface and that it is found in the proper location to bind to the zona pellucida.

Acrosome↗

PubMed1997

Two graduating master's students struggle to find meaning.

This article was initiated to help two graduating Master's students learn what might be expected of them upon graduation. The purpose of this article is to provide insight from reviewing the current literature on clinical nurse specialist's (CNS's) characteristics. The authors believe that this information is especially useful for graduate CNS students and helpful for those who currently hold CNS positions. Based on a literature review, the article categorizes descriptions of the characteristics of the CNS role as perceived by the CNS, management, staff nurses, and physicians. Within the identified perceptions of each group were the following similarly common CNS role components: (a) clinical practice, (b) education, (c) administration, (d) research, and (e) consultation. The key to congruent expectations and understanding of the CNS role is clear communication between the members of each group. Without this communication, the common result is ambiguity, conflict, and frustration.

Hospital Administrators↗

PubMed1996

Effects of a multidimensional anabolic steroid prevention intervention. The Adolescents Training and Learning to Avoid Steroids (ATLAS) Program.

OBJECTIVE: To test a team-based, educational intervention designed to reduce adolescent athletes' intent to use anabolic androgenic steroids (AAS). DESIGN: Randomized prospective trial. SETTING: Thirty-one high school football teams in the Portland, Ore, area. PARTICIPANTS: Seven hundred two adolescent football players at experimental schools; 804 players at control schools. INTERVENTION: Seven weekly, 50-minute class sessions were delivered by coaches and student team leaders, addressing AAS effects, sports nutrition and strength-training alternatives to AAS use, drug refusal role play, and anti-AAS media messages. Seven weight-room sessions were taught by research staff. Parents received written information and were invited to a discussion session. MAIN OUTCOME MEASURES: Questionnaires before and after intervention and at 9- or 12-month follow-up, assessing AAS use risk factors, knowledge and attitudes concerning AAS, sports nutrition and exercise knowledge and behaviors, and intentions to use AAS. RESULTS: Compared with controls, experimental subjects at the long-term follow-up had increased understanding of AAS effects, greater belief in personal vulnerability to the adverse consequences of AAS, improved drug refusal skills, less belief in AAS-promoting media messages, increased belief in the team as an information source, improved perception of athletic abilities and strength-training self-efficacy, improved nutrition and exercise behaviors, and reduced intentions to use AAS. Many other beneficial program effects remained significant at the long-term follow-up. CONCLUSIONS: This AAS prevention program enhanced healthy behaviors, reduced factors that encourage AAS use, and lowered intent to use AAS. These changes were sustained over the period of 1 year. Team-based interventions appear to be an effective approach to improve adolescent behaviors and reduce drug use risk factors.

Adolescent↗

PubMed1996

Nucleotide sequence of ub52 from the cnidarian Acropora millepora reveals high evolutionary conservation.

We report the cDNA sequence encoding Ub52 from the cnidarian Acropora millepora. As in other eukaryotes, the N-terminal region of the deduced amino-acid sequence is a ubiquitin moiety and the C-terminal region CEP52, a protein component of the large ribosomal subunit. A millepora Ub52 is highly homologous (> 95% identical) with the corresponding Drosophila and vertebrate sequences, the cnidarian sequence having higher identity with these 'higher' metazoan homologs than does the corresponding Caenorhabditis elegans protein. However, the cnidarian sequence contains two unique substitutions in the ubiquitin moiety, Ser22 and Thr28. Northern analysis indicates that A. millepora Ub52 is encoded by a small (< 800 nt) mRNA present in eggs and is first expressed after the early gastrula stage in embryonic development.

Amino Acid Sequence↗

PubMed1996

The dipeptide carnosine constricts rabbit saphenous vein as a zinc complex apparently via a serotonergic receptor.

1. The endogenous dipeptide carnosine (beta-alanyl-L-histidine), at 0.1-10 mM, provokes sustained contractures in rabbit saphenous vein rings with greater efficacy than noradrenaline (NA). 2. The effects of carnosine are specific; anserine and homocarnosine are ineffective, as are carnosine's constituent amino acids histidine and beta-alanine. 3. Maximum carnosine-induced tension is enhanced by Zn ions (e.g. to 127.5 +/- 13.1% of control at 10 microM total Zn concentration, Zntot) and the sensitivity to carnosine potentiated (mean [carnosine] required for half-maximal tension, K1/2, reduced from 1.23 mM to 17.0 microM carnosine with 15 microM Zntot). 4. The dipeptide apparently acts as a zinc-carnosine complex. The effects of carnosine at concentrations of 1 microM to 10 mM in the presence of 1-100 microM Zntot, can be described as a unique function of the concentration of Zn-carnosine, with an apparent K1/2 for the complex of 7.4 x 10(-8) M. 5. Contractures are reduced at low [Ca2+], unaffected by adrenoceptor antagonists, but can be blocked by serotonergic receptor antagonists including ketanserin and methiothepin. 6. Competition between albumin and carnosine for Zn ions, as might occur in plasma, can be demonstrated experimentally. 7. The mode of action of carnosine is virtually unique: a vascular muscle receptor apparently transduces the action of a dipeptide in the form of a metal chelate.

Adrenergic alpha-Antagonists↗

PubMed1996

cDNAs from Onchocerca sp. encoding members of the MRS3/MRS4 class of mitochondrial solute carriers.

cDNA clones from the parasitic nematodes Onchocerca volvulus and Onchocerca gibsoni encode homologs of the yeast proteins MRS3 and MRS4. Together with an uncharacterised ORF on chromosome III of Caenorhabditis elegans, these constitute a new class of proteins belonging to the mitochondrial solute carrier protein superfamily. So far, five other members of this protein family have been identified in C. elegans, but levels of identity between these and the Onchocerca proteins were considerably lower. Consideration of cysteine content and overall charge implies that the natural substrates of the nematode proteins are small ions.

Adenosine Diphosphate↗

PubMed1996

Monoclonal autoantibody SCH94.03, which promotes central nervous system remyelination, recognizes an antigen on the surface of oligodendrocytes.

A monoclonal antibody SCH94.03, made in syngeneic mice by injection of spinal cord homogenate, promotes central nervous system remyelination when injected into SJL/J mice chronically infected with Theiler's virus. To elucidate the mechanism of antibody-mediated remyelination, SCH94.03 was investigated by immunocytochemistry, flow cytometry, immunoelectron microscopy, Western blotting, and immuno-thin layer chromatography. All cell types investigated in vitro showed strong cytoplasmic staining with a pattern resembling a cytoskeletal protein. In contrast, among the primary cultured cells studied, only oligodendrocytes showed strong surface reactivity. Other cell types, including astrocytes, microglia, Schwann cells, myoblasts, and T and B lymphocytes, were negative. Mouse and rat oligodendrocytes which showed strong surface reactivity exhibited a well-differentiated morphology, and approximately 50% expressed myelin basic protein. Since oligodendrocyte progenitors were negative for surface staining, the expression of the antigens recognized by this monoclonal antibody appears to be developmentally regulated, i.e., transiently expressed on younger, terminally differentiating oligodendrocytes. Among the cell lines studied, only two rat oligodendrocyte lineage cell lines showed surface reactivity with SCH 94.03. Western blotting of secondary isolated oligodendrocytes lysates revealed reactivity with multiple protein bands of 27, 32, 50, 100, and 106 kDa, whereas there was no reactivity to lipid antigens by immuno-thin layer chromatography. These results raise the possibility that SCH94.03 recognizes a novel oligodendrocyte-specific surface antigen, and may act directly on oligodendrocytes to promote remyelination.

Animals↗

PubMed1996

Spontaneous central nervous system remyelination in strain A mice after infection with the Daniel's (DA) strain of Theiler's virus.

Intracerebral infection of susceptible SJL/J (H-2s) mice with the Daniel's strain of Theiler's murine encephalomyelitis virus produces chronic, progressive, inflammatory central nervous system demyelination, with minimal spontaneous remyelination. To assess the role of host genetic factors in spontaneous myelin repair following chronic infection with the Daniel's strain of Theiler's virus, we examined demyelination and spontaneous remyelination in strain A mice after infection with Theiler's virus. We found that A.BY/SnJ (H-2a), and A.SW/SnJ (H-2s) mice all developed chronic demyelination with substantial spontaneous remyelination 90 days after infection. In the spinal cords of both A/J and A/WySnJ mice, one quarter of the total lesion area showed spontaneous remyelination, whereas in A.SW/SnJ mice, the extent of remyelination increased to two thirds of the total lesion area. The spontaneous remyelination seen in strain A mice was consistent with myelin repair by oligodendrocytes and Schwann cells, and occurred despite the presence of persistent virus antigen. These results indicate that host-pathogen interactions play an important role in myelin regeneration after virus-induced demyelination, and suggest that host genetic factors influence spontaneous remyelination.

Animals↗

PubMed1996

A monoclonal natural autoantibody that promotes remyelination suppresses central nervous system inflammation and increases virus expression after Theiler's virus-induced demyelination.

We have used an established experimental model of multiple sclerosis to investigate the potential beneficial relationship between natural autoimmunity and remyelination after central nervous system (CNS) demyelination. Intracerebral infection of SJL/J mice with Theiler's murine encephalomyelitis virus (TMEV) produces chronic, progressive, inflammatory CNS demyelination. Chronically infected SJL/J mice show minimal spontaneous remyelination, which is in part due to a T cell-mediated immune response inhibiting myelin repair. We previously identified a monoclonal natural autoantibody, designated SCH94.03, that promotes remyelination when passively transferred to chronically infected SJL/J mice. The mechanism whereby SCH94.03 promotes remyelination is unknown, although previous reports suggest that natural autoantibodies can modulate immune system function. In this report we demonstrate that treatment with SCH94.03 reduced by 2- to 3-fold the number of CD4(+) and CD8(+) cells infiltrating the CNS of SJL/J mice chronically infected with TMEV, in the absence of global lymphocyte depletion. Associated with the decreased inflammation was a 2- to 3-fold increase in virus antigen expression without a significant increase in viral RNA or virus titers. Treatment with SCH94.03 also suppressed the humoral immune response to a T cell-dependent antigen in chronically infected mice. Immunohistochemical staining showed that SCH94.03 labeled MHC class II-positive dendritic cells in peripheral lymphoid organs. These results are consistent with the proposed immunomodulatory function of natural autoantibodies and suggest that one mechanism whereby SCH94.03 promotes CNS remyelination in chronically infected SJL/J mice is through inhibition of a pathogenic immune response.

Animals↗

PubMed1996

Central nervous system remyelination clinical application of basic neuroscience principles.

Studies in both humans and experimental animals have demonstrated that myelin repair in the CNS is a normal physiological response to myelin damage, similar to tissue injury elsewhere in the body. The unanswered question is why myelin repair is incomplete in multiple sclerosis patients. In this paper we review the morphological characteristics of remyelination, discuss the available animal models of CNS demyelination and their usefulness to identify the molecular, cellular, and morphological events involved in CNS myelin repair, examine the use of immunosuppression, immunoglobulins, protein growth factors, and glial cell transplantation at the primary experimental therapies designed to promote CNS remyelination, and address the potential electrophysiological and clinical benefits of myelin repair in the CNS.

Animals↗

PubMed1996

Enhanced SR function in saponin-treated ventricular trabeculae from rabbits with heart failure.

Cardiac sarcoplasmic reticulum (SR) Ca(2+)-loading ability was assessed in a coronary artery ligation model of heart failure. Heart failure was produced in New Zealand White rabbits by ligation of the left marginal coronary artery. Sham-operated animals were used as controls. After hemodynamic and echocardiographic assessment 8 wk after coronary ligation, a free-running trabecula was isolated from the left or right ventricle, mounted for isometric tension measurement, and permeabilized with the chemical skinning agent saponin, leaving the SR functionally intact. The SR was Ca2+ loaded by exposure of the preparation to a mock intracellular solution with a Ca2+ concentration ([Ca2+]) of 150-300 nM. The amplitude of the caffeine-induced contracture was used as a measure of Ca2+ loaded by the SR. The same preparation was then treated with Triton X-100 to disrupt all cell membranes, and Ca2+ sensitivity {expressed as [Ca2+] required to produce 50% of maximal activation (pCa50)} of isometric tension production and maximum Ca2+ activated force (Cmax) were measured. Ligated animals demonstrated enhanced SR Ca(2+)-loading ability that correlated with the degree of left ventricular dysfunction. Enhanced SR Ca2+ loading was associated with evidence of SR Ca2+ overload revealed as spontaneous tension oscillations. Cmax and pCa50 were not significantly different from controls. Increased SR Ca(2+)-loading ability may predispose the SR to Ca2+ overload and could contribute to both contractile dysfunction and arrhythmogenesis in heart failure.

Actin Cytoskeleton↗

PubMed1996

Multi-organ reactivity of a monoclonal natural autoantibody that promotes remyelination in a mouse model of multiple sclerosis.

A contemporary view of autoimmunity suggests that self-reactivity is a normal phenomenon, in contrast to the classical association between autoimmunity and immunopathology. We have previously demonstrated that monoclonal antibody SCH94.03, a natural autoantibody with polyreactivity towards several purified protein and hapten antigens, promotes central nervous system remyelination when passively transferred to SJL/J mice chronically infected with Theiler's murine encephalomyelitis virus, an established experimental model of multiple sclerosis. In this study we characterized the autoreactivity of SCH94.03 with endogenous mouse tissue using immunoperoxide and multiple-color immunofluorescence staining techniques on frozen tissue sections. Within the nervous system, SCH94.03 labeled fibrous astrocytes, ependymal cells, ganglion satellite cells, and a sub-population of microglia, oligodendrocytes, and peripheral nervous system neurons. Outside the nervous system, SCH94.03 labeled gastrointestinal tract smooth muscle and luminal epithelium, erythrocytes, and interdigitating dendritic cells in peripheral lymphoid organs. These data indicate that SCH94.03 is a multi-organ reactive autoantibody and support the hypothesis that autoantibodies can have a beneficial rather than a pathogenic function in central nervous system demyelinating diseases.

Animals↗