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Biomedical subjects

R A Miller

Publications and source records attributed to R A Miller.

At least 73 records · Page 4Linked to original sources
PubMed2000

Differential longevity in mouse stocks selected for early life growth trajectory.

Small body size is associated with superior longevity in several intraspecies comparisons, including dogs bred for specific forms of work, mice and rats fed diets low in calories, rats fed diets low in methionine, and mutant mice whose levels of growth hormone and thyroid hormone are atypically low. To further investigate the interactions among body size, genetic endowment, and longevity, we measured the life span of female mice selectively bred from Institute for Cancer Research stock for differences in rate of body weight gain. These mice were selected for differential rates of growth either early (0-10 days) or later (26-56 days) in the first 2 months of life. The data show a good correlation between the average weight of the stock and its mean longevity, with low body size associated, as predicted, with longer life span. Weight at 3, 6, and 12 months, and weight at peak body weight, are all significant predictors of longevity (among stocks) in univariate regressions; weight at 6 months has the strongest association in stepwise multiple regression. There is no significant correlation between the life span for the stock and the proportion of deaths attributable to neoplasia in this group of mice. The data provide support for the hypothesis that genetic factors that influence early life growth trajectories can have a strong influence on life span. These size-selected mice provide useful tools for analysis of the genetic factors that influence life history parameters, including maturation and aging rates.

Adenocarcinoma↗
PubMed2000

Tibiotalocalcaneal arthrodesis.

The purpose of this multicenter retrospective study of 55 patients (56 ankles) who underwent simultaneous tibiotalocalcaneal arthrodesis with severe disease involving the ankle and subtalar joints was to determine improvement of pain and function. The surgical indications included osteoarthritis, posttraumatic injury, failed previous surgery, talar avascular necrosis, osteoarthritis, and rheumatoid arthritis involving the ankle and subtalar joints. The average age at the time of the operation was 53 years. The average time of follow-up was 26 months after the operation. Fusion was achieved in 48 ankles, with an average time of fusion of 19 weeks. Forty-eight of the 55 patients were satisfied with the procedure. The average leg length discrepancy was 1.4 cm. The average amount of dorsiflexion was 2 degrees and plantar flexion was 5 degrees. Following surgery, 42 patients complained of pain, 40 patients required shoe modification or an orthotic device, and 34 patients had a limp. Fourteen patients described their activity as unlimited. Based on the AOFAS evaluation, the patients scored an average of 66 on the ankle-hind foot scale following surgery. The most common complications were nonunion (8 ankles) and wound infection (6 ankles). This study demonstrates that tibiotalocalcaneal arthrodesis is an effective salvage procedure for patients with disease both involving the ankle and subtalar joints.

Adult↗
PubMed2000

A biomechanical evaluation of internal fixation for ankle arthrodesis comparing two methods of joint surface preparation.

This study compared the biomechanics of two methods of joint preparation for ankle fusion with internal fixation. In the first method, the articular cartilage of the ankle joint was removed leaving the joint contour intact. In the second method, parallel flat saw cuts were used to prepare the joint surface for fusion. The specimens were loaded submaximally in a cyclic repetitive fashion before they were loaded to failure. Increasing joint motion occurred as the number of cycles increased. The difference between the two specimen types was not statistically significant during the repetitive tests or when loaded to failure. These results demonstrate both techniques of joint preparation studied are useful and biomechanically sound.

Adult↗
PubMed2000

Achilles tendon rupture associated with ankle fracture.

The case of a 40-year-old man who sustained a medial malleolar fracture with extension of the fracture into the tibial plafond is discussed. Before surgery, the physical examination revealed an Achilles tendon rupture. Surgical treatment to repair the bone and tendon injury was performed. Achilles tendon rupture is not an uncommon injury, but it is rarely associated with a fracture. When a fracture is present, the Achilles tendon injury can be overlooked, which may result in a delay of treatment or residual morbidity.

Achilles Tendon↗
PubMed2000

Experience using a programmable rules engine to implement a complex medical protocol during order entry.

WizOrder, Vanderbilt University Medical Center's (VUMC) clinician order entry system, is an excellent platform for delivering high-quality decision support to clinical end-users. A scripting language designed to make it easy for non-programmer domain experts to enter rules helps distribute the generation and maintenance of the knowledge-base necessary to drive effective decision support. Domain-experts have used this system to successfully implement relatively simple protocols. The VUMC Care Improvement Committee identified diagnosis of and treatment for suspected or confirmed deep venous thrombosis or pulmonary embolism as an area where decision-support could improve clinician compliance with established evidence-based protocols. The authors describe our experience with using our existing scripting system to implement decision support for a complex medical protocol.

Clinical Protocols↗
PubMed1999

Aging impairs induction of cyclin-dependent kinases and down-regulation of p27 in mouse CD4(+) cells.

To define the link between the early activation defects and the impaired proliferation response of cells from old mice, we characterized the influence of age on expression and activity of proteins that participate in cell-cycle regulation. We found that aging led to significant declines in the ability of mouse CD4(+) T cells to respond to CD3 and CD28 stimuli by induction of the cyclin-dependent kinases CDK2, CDK4, and CDK6, whether the defect was assessed by protein level or functional activity. Induction of CDK2 activity was also impaired in cells from old mice that were activated with PMA plus ionomycin, stimuli that bypass the TCR/CD3 complex, or by CD3/CD28 in the presence of IL-2, indicating that the age-related changes lie, at least in part, downstream of the enzymes activated by these stimuli. We also noted an impairment in the ability of CD4(+) cells from old mice to down-regulate the CDK inhibitor p27 after activation, but we found no change in induction of p21, an inhibitor of CDK that may also play other roles in cell-cycle control. Altered CDK activation is likely to mediate the age-related decline in T cell proliferation to polyclonal stimulation.

Aging↗
PubMed1999

Age-related decline in activation of JNK by TCR- and CD28-mediated signals in murine T-lymphocytes.

c-Jun N-terminal kinase (JNK) is activated when T-lymphocytes are stimulated jointly through the T-cell receptor (TCR) and CD28, and it contributes to T-cell activation and IL-2 production through phosphorylation of transcription factors, including c-Jun. We performed in vitro kinase assays on JNK in CD4(+) T-cells, from young and old mice, activated by antibodies to CD3, CD4, and CD28, and found a approximately 2-fold decline in JNK activity at the peak of activation, but no significant change in the kinetics of stimulation or in the steady-state expression of JNK. We found a similar decline in c-Jun phosphorylation in stimulated CD4(+) T-cells from old mice, suggesting that JNK activation also declined with age in intact cells. Aging does not, however, alter the level of Ras activation by anti-CD3/CD4 +/- anti-CD28 or change the level of Ras protein in CD4(+) cells, suggesting that the JNK defect is due to changes in the regulation of other upstream regulators. Our results suggest that a decline with age in JNK responses may contribute to the decline in proliferation and IL-2 production seen in CD4(+) T-cells from old mice.

Aging↗
PubMed1999

Age-sensitive and -insensitive pathways leading to JNK activation in mouse CD4(+) T-cells.

The c-Jun N-terminal kinase (JNK) can be activated in T-cells either by the combination of TCR and CD28 costimulation or by a variety of stress-related stimuli including UV light, H(2)O(2), and hyperosmolar sorbitol solutions. In T-lymphocytes, TCR/CD28 stimulation of JNK leads to induction of new gene expression via c-Jun, ATF-2, and Elk-1. Phosphorylation of c-Jun in CD4(+) T-cells stimulated by CD3/CD4/CD28 cross-linking declines with age, due to diminished activation of JNK. Here we show that the age-related decline in TCR/CD28 activation of JNK reflects two effects of age: the accumulation of memory cells (in which JNK stimulation is poor regardless of donor age) and age-dependent declines in JNK activation within the naive subset. Cyclosporin A inhibits induction of JNK function by TCR/CD28, PMA/ionomycin, ceramide, or H(2)O(2), but not induction by UV light or hyperosmolar sorbitol. Although aging impairs JNK induction by UV light, it has no effect on JNK activation by ceramide, H(2)O(2), or sorbitol. The data as a whole indicate that there are at least four pathways that activate JNK in CD4(+) T-cells, of which two are age-sensitive and two others unaffected by aging. Two of the pathways (UV and hyperosmolar sorbitol) are insensitive to cyclosporin inhibition. Finally, we show that the alterations in JNK function are not due to changes in the expression of MKK4, an upstream activator of JNK, and that another JNK kinase, MKK7, is not expressed in splenic T-cells.

Aging↗
PubMed1999

In vivo animal studies with gadolinium (III) texaphyrin as a radiation enhancer.

PURPOSE: Gadolinium texaphyrin (Gd-Tex, PCI-0120) is an expanded porphyrin that has demonstrated radiation enhancement. In this study, we evaluated the radiation enhancement and biolocalization of Gd-Tex in three animal tumor models. METHODS AND MATERIALS: EMT6, SMT-F, and MCa tumors were established intramuscularly or subcutaneously. Gd-Tex and other metallotexaphyrins were administered prior to single or multiple fractions of radiation. 14C-labeled Gd-Tex was used for biolocalization studies. RESULTS: Gd-Tex, in combination with radiation, produced significant tumor growth delay compared to irradiated control groups in both single and multifraction radiation studies. Gd-Tex radiation enhancement was observed only when the drug was given before, but not after irradiation. Several metallotexaphyrins, identical except for the metal ion, were studied in the EMT6 tumor model including gadolinium (Gd), lutetium (Lu), europium (Eu), yttrium (Y), and cadmium (Cd) texaphyrin complexes. Only Gd-Tex produced radiation enhancement. Biodistribution studies using 14C-labeled Gd-Tex demonstrated drug selectivity and retention in tumors growing intramuscularly compared to uninvolved muscle and plasma. CONCLUSIONS: Gd-Tex produces reproducible radiation enhancement in a variety of in vivo tumor models. This drug's unique radiochemistry, tumor selectivity, and in vivo activity suggests possible mechanisms of action not addressed by in vitro assay methods.

Animals↗
PubMed1999

mdr1a-encoded P-glycoprotein is not required for peripheral T cell proliferation, cytokine release, or cytotoxic effector function in mice.

The plasma membrane transport protein P-glycoprotein (P-gp) is expressed by subsets of both CD4+ and CD8+ T cells in mice. The proportion of T cells that express P-gp goes up with age, and the P-gp-expressing subset of the CD4 memory population is hyporesponsive in many in vitro assays. The significance of P-gp expression for T cell function has not been well established, although several reports have suggested that it may promote cytokine export and/or cytotoxic T cell function. To elucidate which T cell functions may require P-gp, we have compared a variety of responses using T cells from wt and P-gp knockout mice. Protein expression and rhodamine-123 efflux studies revealed that peripheral T cells exclusively utilize the mdr1a-encoded isoform rather than the homologous mdr1b or mdr2 isoforms. Comparisons of T cells from mdr1a+/+ and mdr1a-/- mice showed no differences in proliferation or in secretion of IL-2, IL-4, IL-5, IL-10, or IFN-gamma in response to polyclonal stimulation. Moreover, mdr1a-/- T cells produced strong allospecific cytotoxic responses comparable to those of wt T cells. Our results show that P-gp is not a necessary component of peripheral T cell functional responses. Further investigation will be needed to determine the significance of P-gp expression in T lymphocytes.

ATP Binding Cassette Transporter, Subfamily B↗
PubMed1999

Empirical derivation of an electronic clinically useful problem statement system.

Problem lists are tools to improve patient management. In the medical record, they connect diagnoses to therapy, prognosis, and psychosocial issues. Computer-based problem lists enhance paper-based approaches by enabling cost-containment and quality assurance applications, but they require clinically expressive controlled vocabularies. Because existing controlled vocabularies do not represent problem statements at a clinically useful level, we derived a new canonical problem statement vocabulary through semi-automated analysis and distillation of provider-entered problem lists collected over 6 years from 74,696 patients. We combined automated and manual methods to condense 891,770 problem statements entered by 1961 care providers at Grady Memorial Hospital in Atlanta, Georgia, to 15,534 Canonical Clinical Problem Statement System (CCPSS) terms. The nature and frequency of problem statements were characterized, interrelations among them were enumerated, and a database capturing the epidemiology of problems was created. The authors identified 23,503 problem relations (co-occurrences, sign-symptom complexes, and differential diagnoses) and 22,690 modifier words that further categorized "canonical" problems. To assess completeness, CCPSS content was compared with that of the 1997 Unified Medical Language System Metathesaurus (containing terms from 44 clinical vocabularies). Unified Medical Language System terms expressed 25% of individual CCPSS terms exactly (71% of problems by frequency), 27% partially, and 48% poorly or not at all. Clinicians judged that CCPSS terms completely captured their clinical intent for 84% of 686 randomly selected free-text problem statements. The CCPSS represents clinical concepts at a level exceeding that of previous approaches. A similar national approach could create a standardized, useful, shared resource for clinical practice.

Humans↗
PubMed1999

Cluster formation by protein kinase Ctheta during murine T cell activation: effect of age.

Protein kinase Ctheta; (PKCtheta;) is thought to play an important role in T cell activation, in that exposure of cloned T cells to antigen-presenting cells bearing agonist peptides, but not antagonist peptides, leads to clustering of PKCtheta; molecules in the section of the T cell plasma membrane that is in contact with the APCs. To see whether aging affects this PKCtheta; clustering reaction in mouse T lymphocytes, we used immunofluorescence staining and confocal microscopy to observe the localization of PKCtheta; in CD4 and CD8 T lymphocytes activated by coincubation with anti-CD3 hybridoma cells. Aging led to a twofold decline in the proportion of both CD4 and CD8 T cells in which PKCtheta; underwent cluster formation. This decrease with age was not due to differences in the number of cell conjugates formed, nor to kinetic differences of PKCtheta; clustering, nor to the accumulation of memory T cells in old mice. There were no effects of aging on the levels or kinase activity of PKCtheta; in murine T cells. Our data suggest alterations in the upstream signals that regulate PKCtheta; translocation.

Aging↗
PubMed1999

Calcium signal abnormalities in murine T lymphocytes that express the multidrug transporter P-glycoprotein.

Previous work has established that aging in mice leads to an accumulation of T cells that express high levels of P-glycoprotein, a plasma membrane pump that mediates multiple drug resistance in tumor cells but whose function in normal T cells is still obscure. Pgp+ cells seem to be functionally defective: isolated from the CD4 memory population of young mice, they are unresponsive to T cell receptor-dependent stimuli in tests for proliferation and cytokine production. The proliferative defect can, however, be overcome by exposure to PMA plus the calcium ionophore ionomycin, suggesting that the Pgp+ cells may have a specific defect in calcium signal generation. We show here that Pgp+ T cells, from young or old mice, do indeed show smaller changes in intracellular calcium ion concentration than Pgp- cells, when activated either by Con A, anti-CD3 antibodies, or ionomycin. The difference between Pgp+ and Pgp- cells is apparent even in experiments on isolated CD4 memory T cells from young mice and thus is not simply a consequence of the age-dependent increase in memory cell numbers. Although the molecular basis for the abnormality in calcium signal generation by Pgp+ cells is still uncertain, our data suggest that the effect could be due to inter-subset differences in levels of sorcin, a 22 kDa cytoplasmic protein that is co-expressed with P-glycoprotein in many tumor cells and which binds free calcium ion with high affinity. Sorcin levels are higher in Pgp+ CD4 cells than in Pgp- CD4 cells of young mice and increase with age in CD4 cells, consistent with the hypothesis that sorcin interferes with calcium signals in the age-sensitive Pgp+ T cell subset.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1999

Aging increases CD8 T cell apoptosis induced by hyperstimulation but decreases apoptosis induced by agonist withdrawal in mice.

Apoptosis of T cells is thought to play a critical role in intrathymic T cell selection, in controlling the strength of the immune response to antigens, and in peripheral modulation of the T cell repertoire by influencing memory cell formation and survival. Peripheral T lymphocyte apoptosis or activation-induced cell death can be induced in vitro by repeated stimulation through the T cell receptor (TCR), and several groups have reported that aging increases the susceptibility of T cells to hyperstimulation-induced cell death in mice and humans. Alternately, apoptosis can also be induced in T cells by withdrawal of TCR stimulation from T cell blasts late in the activation process. This agonist withdrawal cell death, unlike apoptosis induced by repeated stimulation, is Fas- and TNFalpha-independent but is modulated by CD30 ligation. We show here that aging leads to an increase in susceptibility to apoptosis induced by repeated stimulation, but also to a decline in mouse CD8 T cell sensitivity to apoptosis induced by agonist withdrawal. Cell mixture experiments show that intercellular signals are required for the induction of apoptosis after agonist withdrawal and that the CD8 cells from aged mice can respond to these death-inducing signals but cannot produce them. A defect in this form of apoptosis after cessation of TCR signaling might contribute to the accumulation of functionally ineffective CD8 cells in aging mice.

Aging↗
PubMed1999

Age-associated decline in responses of naïve T cells to in vitro immunization reflects shift in glucocorticoid sensitivity.

Naïve T lymphocytes from young mice can be immunized to protein antigens in vitro if the initial exposure to antigen is followed by a brief period of clonal expansion in the presence of both the glucocorticoid dexamethasone (at 10(-8) M) and antibodies to Interleukin-10 (IL-10). These cultures produce cell lines that respond to antigen rechallenge by proliferation and cytokine secretion. T cells from older mice, however, do not respond under these conditions unless the dexamethasone concentration is raised to levels (10(-7) M) that are inhibitory for T cells of young mice. Suitably timed exposure to dexamethasone can also increase proliferative responses to polyclonal activation via the CD3 component of the T cell receptor, and again optimal responses are obtained from old mice only at steroid concentrations that are super-optimal for young T cells. Diminished sensitivity to glucocorticoid effects may contribute to the poor responses of aged mice to novel immunogens.

Aging↗
PubMed1999

Development and validation of a high-performance liquid chromatography tandem mass spectrometry assay for atorvastatin, ortho-hydroxy atorvastatin, and para-hydroxy atorvastatin in human, dog, and rat plasma.

A liquid chromatographic/mass spectrometric method to quantitate atorvastatin (AT) and its active metabolites ortho-hydroxy (o-AT) and para-hydroxy (p-AT) atorvastatin in human, dog, and rat plasma was validated. The method consisted of washing plasma samples at high pH with diethyl ether and subsequently extracting the analytes and two internal standards, [d5]-atorvastatin ([d5]-AT) and [d5]-ortho-hydroxy atorvastatin ([d5]-o-AT), from acidified plasma by using diethyl ether. The ether layer was evaporated to dryness and the residue reconstituted in ammonium acetate (20 mM, pH 4.0)-acetonitrile-isopropanol (60:40:1, v/v/v). Chromatographic separation of analytes was achieved by using a YMC J'Sphere H80 (C-18) 150 x 2 mm, 4 microns particle size, column with a mobile phase consisting of acetonitrile-0.1% acetic acid, (70:30, v/v). Analytes were detected by using MS/MS. Sample introduction and ionization was by electrospray ionization in the positive ion mode. The method proved suitable for routine quantitation of AT, o-AT, and p-AT over the concentration range of 0.250 to 25.0 ng/mL. Approximate retention time ranges of p-AT, o-AT, [d5]-o-AT, AT, and [d5]-AT were 2.27 +/- 0.21, 3.36 +/- 0.23, 3.54 +/- 0.46, 4.12 +/- 0.61, and 4.65 +/- 0.65 min, respectively. No peaks interfering with quantitation were observed throughout the validation processes. Mean recoveries of AT, o-AT, and p-AT from plasma ranged 100%-107%, 70.6%-104%, and 47.6%-85.6%, respectively. Mean recoveries of the [d5]-AT and [d5]-o-AT internal standards ranged 98.0%-99.9% and 97.3%, respectively. Interassay precision, based on the percent relative deviation for replicate quality controls for AT, o-AT, and p-AT, was < or = 7.19%, 8.28%, and 12.7%, respectively. Interassay accuracy for AT, o-AT, and p-AT was +/- 10.6%, 5.86%, and 15.8%, respectively. AT, o-AT, and p-AT in human, dog, and rat plasma quality controls were stable to three freeze-thaw cycles. AT, o-AT, and p-AT were stable frozen for 127, 30 and 270 days in human, dog, and rat plasma quality control samples, respectively. Human plasma quality control samples containing AT, o-AT, and p-AT were stable for at least 4 days at ambient room temperature and 37 degrees C. The lower limit of quantitation for all analytes was 0.250 ng/mL for a 1.0-mL sample aliquot.

Animals↗
PubMed1999

The compression of spheres coated with an aqueous ethylcellulose dispersion.

Tablets were compressed from commercial samples of Sugar Spheres NF, Sucrose NF, Corn Starch NF, as well as ground spheres and a physical mixture of ground sucrose plus cornstarch. Additional tablets were compressed from spheres that had been coated with a water-soluble cellulosic polymer solution followed by an aqueous ethylcellulose dispersion. Tableting parameters measured "in-die" included work of compression, peak offset time, tablet density, and Young's modulus. Following ejection, tensile strength was determined under diametrical loading. Dissolution of a marker contained in the water-soluble layer was determined for both compressed and uncompressed spheres. Porosities at peak pressure and peak offset times or tensile strength as functions of peak pressure did not differ between tablets compressed from pristine spheres or from ground spheres. Tablets compressed from spheres had higher values for porosity, tensile strength, and peak offset time than those compressed from sucrose or the sucrose: starch mixture. Values for work of compression were higher for tablets compressed from pristine spheres or from starch. This was attributed to the work required for particle deformation and for breaking of the spheres. The greatest elastic recovery during decompression was observed for tablets compressed from pristine spheres or starch. More brittle behavior was observed for tablets compressed from sucrose or the sucrose: starch mixture. Tablets compressed from ground spheres were more brittle than those compressed from the pristine spheres, indicating an effect due to grinding. Most mechanical properties of tablets compressed from the coated spheres were comparable to those of tablets compressed from uncoated spheres. An exception was diametric strain for tablets compressed from spheres coated with the aqueous ethylcellulose dispersion. These values increased since the plasticized ethylcellulose allowed greater distortion of the tablet before failure occurred. The dye marker was released more rapidly from tablets compressed from spheres coated with the aqueous ethylcellulose dispersion than from comparable uncompressed spheres. At both the 5% and 10% coating levels, spheres coated with the aqueous ethylcellulose dispersion fused into nondisintegrating matrices during compression. Little difference in release rates was seen between the two tablets.

Carbohydrates↗