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Biomedical subjects

M L Miller

Publications and source records attributed to M L Miller.

At least 91 records · Page 5Linked to original sources
PubMed1995

Relationships between the nuclear membrane, nuclear pore complexes, and organelles in the type II pneumocyte.

Functional relationships among organelles of the type II cell are suggested based upon the proximity of organelles to specialized areas of the plasma- and nuclear membranes. In a two-dimensional morphometric analysis of the profiles of organelles in type II cells of the ferret and rat (and beagle dog), lamellar bodies were more likely to be located near the nuclear membrane than at the alveolar space (where exocytosis occurs). The size of lamellar body profiles was not correlated with distance from the nuclear membrane; however, large profiles were nearer the apical membrane, and smaller ones nearer the basement membrane. Profiles of highly branched mitochondria were 10 times more frequently associated with nuclear pore complexes than with the inter-pore nuclear membrane. Forty percent of all mitochondrial profiles were within 0.25 microns of the nucleus, 5% were within 0.02 microns and half of these appeared to touch the filaments of the nuclear pore complexes. The size of mitochondrial profiles was not correlated with distribution. In the ferret and rat, 8.6% and 2.5% respectively, of the nuclear pore complexes were associated with mitochondria. Sebaceous cells, from control mice, demonstrated a spatial distribution of granules which was size dependent but unrelated to the nuclear membrane.

Animals↗
PubMed1995

Transglutaminase cross-linking of the tau protein.

Tissue transglutaminase (EC 2.3.2.13) is a calcium-activated enzyme that cross-links specific substrate proteins into insoluble, protease-resistant, high molecular weight complexes. Because the neurofibrillary tangles in Alzheimer disease have similar biochemical characteristics, and because the microtubule-associated protein tau is the predominant component of these structures, the substrate properties of tau with respect to transglutaminase were investigated. Bovine tau and recombinant human tau isoforms rapidly form high molecular weight, cross-linked polymers on incubation with transglutaminase. Polyamine incorporation assays indicate that bovine tau is an excellent substrate of transglutaminase, with a Km of 10.4 +/- 2.2 microM and a Vmax of 40.9 +/- 4.5 nmol/mg of enzyme/min. Individual recombinant human tau isoforms are not equivalent with respect to transglutaminase, as the smallest isoform T3 (352 amino acids) is not as good a substrate as the larger isoforms T4 (383 amino acids) and T4L (441 amino acids). To determine which segments of the tau protein are susceptible to modification by transglutaminase, tau was labeled with [3H]putrescine by transglutaminase and proteolyzed with alpha-chymotrypsin, and the breakdown products were analyzed. These experiments demonstrate that the enzyme modifies tau at only one or a few discrete sites, primarily in the carboxyl half of the molecule. Thus, the reaction is specific for only a small number of the many glutamine residues in tau. Furthermore, a tau deletion construct (T264) containing a portion of the microtubule-binding domains, which is a substrate of transglutaminase, cannot be cross-linked by the enzyme. This provides evidence that the cross-linking reaction is specific, and requires that the substrates be appropriately associated for cross-linking to occur.

Alzheimer Disease↗
PubMed1995

The effect of a heparin removal filter on platelet aggregation studies in heparin-induced thrombocytopenia.

Patients having a heparin-associated platelet antibody who are receiving heparin at the time of testing for heparin-induced thrombocytopenia (HIT) by platelet aggregometry may exhibit aggregation in the negative control channel. Filtering plasma to remove the heparin in may produce a nonaggregating negative control channel. The effect of the Pall Hepchek (Pall Biomedical, East Hills, NY) heparin removal filter on platelet aggregation studies was evaluated. Samples were studied from 10 patients with clinically established HIT. The pre-filtration platelet aggregation studies were unequivocally positive for heparin antibody. The remainder of each sample was filtered and the aggregation studies were repeated. Of the four patients on IV heparin, only one remained positive post-filtration. Of the six patients receiving subcutaneous heparin or flushes, one remained strongly positive, one was borderline, and four became negative. Pall Hepchek heparin filtration unpredictably alters the results of platelet aggregation studies, and should not be used routinely to remove heparin in the presence of aggregation in the negative control.

Filtration↗
PubMed1995

Time course of airway hyperresponsiveness and remodeling induced by hyperoxia in rats.

The purpose of this study was to answer two questions concerning hyperoxia-induced airway hyperresponsiveness: 1) What is the time course of the development of airway hyperresponsiveness? 2) What is the relationship between the increase in responsiveness and smooth muscle area? Segments of intrapulmonary bronchi were isolated from male Sprague-Dawley rats that had been exposed to 80-85% O2 for a period of 1, 3, 5, or 7 days and from aged-matched control animals that breathed room air. Hyperoxia increased the sensitivity (log concentration or frequency that elicited a half-maximal response) and reactivity (maximum tension developed) of the airways to electrical field stimulation (EFS) after 3, 5, and 7 days; sensitivity to acetylcholine was not affected, but reactivity was increased after 7 days. Hyperoxia increased smooth muscle area beginning 5 days after commencing the exposure. After normalizing tension responses to smooth muscle area, reactivity of the airways to the stimuli was not different between the two groups, but sensitivity to EFS was still increased. The increase in reactivity observed after 5 and 7 days of exposure can be explained by an increase in smooth muscle area that occurred at these time points. The fact that the sensitivity of the airways to EFS remained increased after normalization, together with the fact that the increase in airway responsiveness after 3 days of exposure occurred at a time when smooth muscle area was not different from control, suggests that mechanisms other than increased smooth muscle area contribute to the development of hyperoxia-induced airway hyperresponsiveness.

Acetylcholine↗
PubMed1995

Refractory ceramic fibers activate alveolar macrophage eicosanoid and cytokine release.

Refractory ceramic fiber has been developed for industrial processes requiring materials with high thermal and mechanical stability. To evaluate the biological activity of this fiber, rat alveolar macrophages were exposed for < or = 24 h to 0-1,000 micrograms/ml of refractory ceramic fiber, crocidolite asbestos, silica (fibrogenic particles), or titanium dioxide (a nonfibrogenic particle), and eicosanoid, tumor necrosis factor-alpha (TNF), and lactate dehydrogenase release were measured. Particle dimensions were determined by electron microscopy. Radioactivity coeluting with leukotriene B4 (LTB4) and immunoreactive LTB4 and TNF release increased after refractory ceramic fiber and were similar in magnitude after asbestos but less than after silica. For example, the total [3H]eicosanoid release increased 3.9-fold after refractory ceramic fiber, 4.6-fold after asbestos, and 8.7-fold after silica. Refractory ceramic fiber and asbestos also have similar particle dimensions (diameter, length, and surface area). Inasmuch as macrophage-derived LTB4 and TNF are potent mediators in inflammatory events, including migration and activation of neutrophils, these findings suggest that refractory ceramic fiber can activate macrophages in vitro to release mediators relevant to in vivo findings of inflammation and fibrotic lung disease in laboratory animals.

Animals↗
PubMed1995

Presence of multiple anti-phospholipid antibody specificities in a pediatric population.

Thrombotic related events are thought to be associated with the presence of anti-phospholipid antibodies (APA). However, the association of anti-cardiolipin antibody is much weaker than the association with antibodies to other phospholipids. Much of the literature equates antiphospholipid antibodies and anticardiolipin antibodies because of the relationship of APA and false positive tests for syphilis. However, recently the presence of antibodies to naturally occurring phospholipids other than cardiolipin have been reported. In fact, some investigators report that antibodies to phosphatidylserine appear to correlate more closely to disease processes than anti-cardiolipin antibodies. We describe here the presence of non-anti-cardiolipin antiphospholipid antibodies in a pediatric population that lack anti-cardiolipin antibodies and demonstrate the association of these antibodies with thrombotic disease. Antibodies to phosphatidic acid were the most prevalent and correlated (p < .001) with thrombotic disease and idiopathic thrombocytopenia purpura. The rank order of prevalence of antibodies to phospholipids was phosphatidic acid, phosphatidylglycerol, phosphatidylinosital, phosphatidylserine, cardiolipin and phosphatidylethanolamine. Antiphospholipid antibodies of the three major sera isotypes were present in the positive sera examined. These descriptive findings suggest that the significance of APA other than anti-cardiolipin antibodies in pediatric patients should be further investigated.

Antibodies, Anticardiolipin↗
PubMed1995

Elements in the 5' flanking sequences of the mouse low-affinity NGF receptor gene direct appropriate CNS, but not PNS, expression in transgenic mice.

We have initiated a characterization of the cis-acting regulatory elements of the murine low-affinity NGF receptor (p75NGFR) gene. Despite studies in cultured cells that suggest the p75NGFR promoter is constitutive, a detailed analysis of this promoter in five lines of transgenic mice demonstrated a high degree of cell-type specificity: 8.4 kb of 5' flanking sequence directs expression of a lacZ reporter to retinal and CNS neurons normally expressing p75NGFR. A transgene with 470 bp of 5' flanking sequence is also expressed in the CNS, but its regulation is aberrant, with a loss of basal forebrain expression. In non-neural tissues, both transgenes were expressed only in the testis, kidney, anterior pituitary, and pancreatic islets; with the exception of the renal pattern of expression, transgene activity was confined to appropriate cells within these tissues. In contrast, although expression of both transgenes was prominent in adrenal medulla and gastrointestinal myenteric neurons, neither construct was active in several sensory or sympathetic ganglia that strongly express the endogenous p75NGFR gene, indicating that genetic elements necessary for expression in these neurons are not present in these promoter sequences. In addition, neither transgene was activated in Schwann cells during Wallerian degeneration of sciatic nerve. We conclude that regulation of the p75NGFR gene is complex, with the first 470 bp of 5' flanking sequence sufficient for expression in enteric and CNS neurons and additional elements within the first 8.4 kb of 5' flanking sequence required for restriction to appropriate CNS neurons. Further regulatory elements are possibly required for expression in at least some sensory and sympathetic neurons in the PNS and in Schwann cells. To identify potential regulatory elements in the 470 bp of 5' flanking sequence from the smaller transgene, we compared the sequences of equivalent regions from the mouse, rat, and human p75NGFR genes. This "phylogenetic footprint" identified conserved motifs potentially important for the regulation of this gene in the CNS.

Animals↗
PubMed1995

Abnormal muscle MRI in a patient with systemic juvenile arthritis.

Although myositis has been described in children with systemic-onset juvenile arthritis (JA), its documentation by MRI has not been reported. We describe a 13-year-old boy with systemic-onset JA, severe myalgia, and elevated muscle enzymes, but normal muscle strength, who had an MRI consistent with myositis. Magnetic resonance imaging can identify the specific location of myositis, allowing more precise definition of a potential complication of systemic JA.

Adolescent↗
PubMed1994

Carcinogenicity of 7H-dibenzo[c,g]carbazole, dibenz[a,j]acridine and benzo[a]pyrene in mouse skin and liver following topical application.

N-Heterocyclic aromatics are by-products of incomplete combustion of organic material. The overall objective of this study was to determine the relative carcinogenic potencies of 7H-dibenzo[c,g]carbazole (DBC) and dibenz[a,j]acridine (DBA) in a bioassay of complete carcinogenicity on mouse skin in a sensitive strain (Hsd:ICR(Br)) which has been used in metabolism and DNA binding studies of N-heterocyclic aromatics. No-treatment, acetone and benzo[a]pyrene (BaP)-treated animals were used as negative and positive control groups. DBC (50 nmol), DBA (50 nmol), BaP (50 nmol) or DBC plus BaP (25 nmol + 25 nmol) were applied twice weekly in 50 microliters acetone to the backs of 50 female mice/group for 99 weeks or until the appearance of a tumor. DBC, DBA, BaP and DBC plus BaP produced skin tumors in 43, 32, 49 and 42 of 50 mice each based on weekly visual observations with latent periods of 55.1, 62.2, 33.4 and 33.8 weeks, respectively. The histopathology data indicated primary skin lesions in 42, 27, 48 and 47 mice for DBC, DBA, BaP and DBC plus BaP, respectively. In addition, primary liver lesions in 37 mice were present in the DBC group. The morphological and morphometric data indicated a significant increase (P < or = 0.05) in mononuclear cells in the dermis for the BaP and DBC plus BaP groups relative to the control group. Significant increases (P < or = 0.05) were observed in the nuclear area, nucleoli per nucleus and cellular area of hepatocytes in the DBC treatment group relative to the control group. These data indicate that DBC is a potent liver carcinogen as well as a skin carcinogen following topical application.

Acridines↗
PubMed1994

Olfactory neurons in vitro show phenotypic orientation in epithelial spheres.

Production and differentiation of olfactory neurons occur in spherical, multi-neuronal aggregates that form in cultures where dissociated newborn rat nasal cells are plated on to CNS glial cells. We show here that neuronal cell bodies were primarily located in the peripheral layers of the spheres, and almost every neuronal sphere contained one or several non-cellular central cavities. The dendrite-like processes of the olfactory neurons, immunostained for neuron-specific tubulin or the olfactory marker protein, were aligned and directed towards the central cavities. Olfactory neurons in the intact animal show a similar relationship with the nasal lumen. Non-neuronal cells formed multiple layers centrally, bordering the cavities. This degree of phenotypic re-creation is unusual in a dissociated monolayer culture system.

Animals↗
PubMed1994

Escape behavior in task situations: task versus social antecedents.

We designed an investigation to differentiate two types of challenging behaviors occurring in teaching situations: those evoked by task stimuli (i.e., task avoidance), and those evoked by social stimuli present in teaching situations (i.e., social avoidance). Four students with developmental disabilities who exhibited challenging behaviors in teaching situations were exposed to social interaction in a play situation and task demands in a teaching situation. Results indicated that the students exhibited two distinct behavior patterns. Two of the students exhibited a behavior pattern consistent with task avoidance and the other two students exhibited a behavior pattern consistent with social avoidance. Implications concerning task versus social avoidance and the need for more fine-grained analyses of the stimuli associated with escape behavior are discussed.

Autistic Disorder↗
PubMed1994

Granulocyte-macrophage colony-stimulating factor (GM-CSF) is expressed in mouse skin in response to tumor-promoting agents and modulates dermal inflammation and epidermal dark cell numbers.

In mouse dorsal skin multistage carcinogenesis models, tumor promotion can be mediated by chemical agents, but also by wounding or abrasion of the epidermis, suggesting that endogenous growth factors mediate this process. Granulocyte-macrophage colony-stimulating factor (GM-CSF) is one such factor that has been reported to be produced by keratinocytes in vitro, and has been suggested both to stimulate keratinocyte proliferation, and also to be a chemoattractant for neutrophils and macrophages. In this study we examined the expression and function of GM-CSF in mouse skin following the application of tumor-promoting agents. Both single and multiple applications of 12-O-tetradecanoylphorbol-13-acetate (TPA) resulted in accumulation of GM-CSF mRNA in the epidermis. Various phorbol and non-phorbol ester tumor promoters were found to induce increases in epidermal GM-CSF mRNA levels commensurate with their relative tumor promoting capabilities. Fluocinolone acetonide (FA) and tosyl phenylalanine chloromethyl ketone (TPCK), inhibitors of tumor promotion, inhibited tumor promoter-mediated GM-CSF accumulation, whereas all-trans-retinoic acid (RA) enhanced the TPA-induced increase. The retinoic acid analogue RO-109359 which, unlike RA, does not have tumor promoting activity per se, inhibited the TPA-induced increase in epidermal GM-CSF mRNA levels. When an antibody specific to GM-CSF was administered prior to TPA, the promoter-induced dermal inflammation and increase in epidermal dark cell number were reduced, yet promoter-induced epidermal hyperplasia was not. These findings implied that elevation of GM-CSF levels plays an important role in chemically-mediated mouse skin tumor promotion and principally via effects on promoter-induced inflammation and increased epidermal dark cell number.

Animals↗
PubMed1994

Induction of interleukin-6 in the epidermis of mice in response to tumor-promoting agents.

Recent reports imply that several epidermal cytokines have a functional role in the tumor promotion stage of the multistage carcinogenesis model in mouse dorsal skin. In this report we describe studies to assess the role of interleukin-6 (IL-6) in tumor promotion. Promoting, as well as non-promoting, hyperplastic agents were found to induce IL-6, as measured by mRNA expression. Inhibitors of tumor promotion inhibited tumor-promoter-mediated IL-6 induction. However, when mice were injected with a neutralizing antibody specific to murine IL-6, there was no effect on tumor-promoter-mediated epidermal hyperplasia and dermal inflammation. These studies suggest that even though IL-6 is produced in the epidermis following tumor promoter application, it does not modulate epidermal hyperplasia and dermal inflammation. Our findings also stress the importance of assessing, in in vivo studies, the function of those cytokines suggested by in vitro experiments to have important roles in tumor promotion.

Animals↗
PubMed1994

Cleanup protocol for 226Ra-contaminated cobbly soil at UMTRA Project sites.

The nonuniform distribution of 226Ra and other radiological contamination of cobbly soil encountered on several Uranium Mill Tailings Remedial Action Project sites is presented and discussed, and the concomitant challenges to the intent and implementation of the U.S. Environmental Protection Agency's soil cleanup standards are noted. In response to technical assessments and information presented to the U.S. Nuclear Regulatory Commission by the U.S. Department of Energy, the Nuclear Regulatory Commission has recently resolved the dilemma by concluding that compliance with Environmental Protection Agency soil cleanup standards for cobby soil at Uranium Mill Tailings Remedial Action Project sites would be adequately attained using bulk radionuclide concentrations, instead of requiring that the radionuclide concentration of the finer soil fraction passing a #4 mesh sieve met the standards. A Nuclear Regulatory Commission-approved procedure developed for cobbly soil remediation is outlined and discussed. The site-specific implementation of this procedure at Uranium Mill Tailings Remedial Action Project sites containing cobbly soil is estimated to save millions of dollars.

Colorado↗