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Biomedical subjects

C Reynolds

Publications and source records attributed to C Reynolds.

At least 19 recordsLinked to original sources

Two crystal forms of the lentil lectin diffract to high resolution.

The legume lectins are an important class of polysaccharide-binding proteins with a wide range of biochemical and immunological applications. Two high-resolution crystal forms are obtained for the lentil (Lens culinaris) lectin: a monoclinic P21 and an orthorhombic P212121. The unit cell dimensions for the monoclinic form are a = 58.0 A, b = 56.0 A, c = 82.1 A, beta = 104.4 degrees, while for the orthorhombic form a = 56.4 A, b = 74.6 A, c = 124.9 A. The asymmetric unit contains one dimer in both cases. The crystals diffract to 1.7 A resolution using synchrotron radiation. Preliminary data have been collected to 2.3 A on both crystal forms using a conventional X-ray source.

Crystallography

Biometrical analysis of individual growth curves.

Longitudinal data for height (length) between birth and 2 years of age were examined for 690 Dutch Registry twin pairs. A two-stage analysis was performed, where individual growth curves were first fit to available data for each subject using a linear multiple regression procedure and estimated individual growth curve parameters were then subjected to multivariate biometrical analysis. Quadratic polynomial curves were found to adequately represent observed growth patterns for the majority of cases (median R2 = .98). A specific scalar model of sex limitation best characterized individual variation in growth curve parameters. That is, there was significantly greater genetic variation for boys than for girls in both the predicted length and rate of growth at 1 year of age and the amount of deceleration in individual growth curves across age.

Anthropometry

Multi-state investigation of the actual disinfection/sterilization of endoscopes in health care facilities.

PURPOSE: The purpose of this investigation was to observe and assess the actual disinfection or sterilization of endoscopes in health care facilities. MATERIALS AND METHODS: A total of 22 hospitals and four ambulatory care centers in three states were studied. Facility protocols were reviewed, interviews conducted with relevant personnel, actual disinfection or sterilization procedures observed, and biologic tests performed to determine and assess disinfection/sterilization procedures. RESULTS: Fundamental errors observed during the course of the investigation included respective failures to time the period of disinfection, to clean all channels, to flush all channels with disinfectant, to fully immerse the endoscope in the disinfectant solution, and to use a disinfectant. At 78% of the facilities, failure to sterilize all biopsy forceps was observed. A total of 23.9% of the bacterial cultures from the internal channels of 71 gastrointestinal endoscopes grew 100,000 colonies or more of bacteria. These cultures were obtained after the completion of all disinfection/sterilization procedures and the device was deemed ready for use in the next patient. CONCLUSIONS: These data indicate that actual disinfection/sterilization procedures for endoscopes are not always optimal, and high-level disinfection of gastrointestinal endoscopes is not always achieved.

Acquired Immunodeficiency Syndrome

Folding and processing of the capsid protein precursor P1 is kinetically retarded in neutralization site 3B mutants of poliovirus.

Poliovirus mutants in neutralizing antigenic site 3B were constructed by replacing the glutamic acid residue at amino acid 74 of capsid protein VP2 (VP2074E), using site-specific mutagenesis methods. All viable mutants display small-plaque phenotypes. Characterization of these mutants indicates that capsid assembly is perturbed. Although the defect in capsid assembly reduces the yield of mutant virus particles per cell, the resultant assembled particle is wild-type-like in structure and infectivity. Analyses of capsid assembly intermediates show a transient accumulation of the unprocessed capsid protein precursor, P1, indicating that cleavage of the mutant P1 by the 3CD protease is retarded. The mutant VP0-VP3-VP1 complex generated upon P1 cleavage appears assembly competent, forming pentamer and empty capsid assembly intermediates and infectious virion particles. Although the structure of the infectious mutant virus is virtually identical with that of the wild-type virus, the thermal stability of the mutant virus is dramatically increased over that of the wild-type virus. Thus, mutations at this residue are pleiotropic, altering the kinetics of capsid assembly and generating a virus that is more thermostable and more resistant to neutralization by the site 3B monoclonal antibodies.

Amino Acid Sequence

Effects of clozapine and thiothixene on glucose metabolic rate in schizophrenia.

Twelve patients with schizophrenia received positron emission tomography scans with 18F-deoxyglucose before and after 4 to 6 weeks of treatment with clozapine or thiothixene. Both stereotaxic and magnetic resonance image template methods were used to position regions of interest for metabolic rate analysis. Clozapine increased and thiothixene decreased metabolic rates in the basal ganglia; these effects were most marked on the right side. Within the basal ganglia, a superior to inferior gradient in drug effect was found for thiothixene but not clozapine. This gradient resembled in some respects observations on regional differences in D2 receptors in human autoradiography. Baseline metabolic rates also predicted clinical medication response, with right inferior caudate metabolic rates differentiating clozapine and thiothixene responders. Larger sample studies are needed to replicate and extend these initial findings.

Adult

The two different receptors for tumor necrosis factor mediate distinct cellular responses.

The individual roles of the murine type 1 and type 2 tumor necrosis factor (TNF) receptors (TNF-R1 and TNF-R2) were investigated utilizing (i) the strong species specificity of TNF-R2 for murine TNF compared to human TNF and (ii) agonistic rabbit polyclonal antibodies directed against the individual TNF receptors. Proliferation of mouse thymocytes and the murine cytotoxic T-cell line CT-6 is stimulated by murine TNF but not by human TNF. Consistent with this observation, polyclonal antibodies directed against TNF-R2 induced proliferation in both of these cell types, whereas polyclonal antibodies directed against TNF-R1 had no effect. In contrast, cytotoxicity in murine LM cells (which are sensitive to murine and human TNF) was induced by antibodies against TNF-R1 but not by antibodies against TNF-R2. Also, the steady-state level of manganous superoxide dismutase mRNA in the murine NIH 3T3 cell line was induced by murine TNF, human TNF, and anti-TNF-R1 but not by anti-TNF-R2. These results suggest that TNF-R2 initiates signals for the proliferation of thymocytes and cytotoxic T cells, whereas TNF-R1 initiates signals for cytotoxicity and the induction of the protective activity, manganous superoxide dismutase. The nonredundant signaling observed for the two TNF receptors cannot be explained simply by the differential expression of the two TNF receptors in the various cell types, because LM cells express on their surface higher levels of TNF-R2 than TNF-R1, and LM cells, NIH 3T3 cells, and thymus cells all express mRNA corresponding to both receptor types. It is therefore likely that the two receptors initiate distinct signaling pathways that result in the induction of different cellular responses.

3T3 Cells

Identification of residues in VP2 that contribute to poliovirus neutralization antigenic site 3B.

Amino acid substitutions were placed at residues 74, 243, and 246 in capsid protein VP2 of poliovirus serotype 1, using site-specific mutagenesis methods. The proximity of these residues to those previously identified in neutralization site 3B suggests that these residues may also contribute to neutralization site 3B and potentially be under antibody selective pressure to mutate. However, sequence analyses of independent serotype 1 isolates indicate high sequence conservation at these residues, suggesting selective pressures are present within the virus to maintain sequences within these loop regions. All viable mutants display partial or complete resistance to neutralization by the site 3B neutralizing monoclonal antibodies. Cross-neutralization data with the site-specifically generated viral mutants confirm that these residues do indeed contribute to forming neutralization site 3B and also identify the participation of a new loop region within site 3B. However, many amino acid substitutions generate nonviable virus mutants and even conservative amino acid substitutions produce growth-compromised virus mutants. These data suggest that previous definition of neutralization antigenic sites by isolation of neutralization resistant mutants favors detection of viable mutant viruses with more normal growth characteristics and is inherently biased against detection of neutralization antigenic sites formed by residues critical for other stages of virus replication.

Amino Acid Sequence

Glove use by health care workers: results of a tristate investigation.

The Center for Devices and Radiological Health, in collaboration with the state health departments of Iowa, Maryland, and Massachusetts, conducted a multi-state, multi-institutional investigation of glove use by health care workers (HCWs). Twenty-two hospitals and four ambulatory care centers were included in the investigation. All 26 health care facilities were found to have adopted universal precautions policies for glove use by HCWs, per Centers for Disease Control guidelines. Four hundred five observations were made of HCWs performing procedures that may involve contact with patient body fluids, particularly blood. The prevalence of glove use during selected procedures was as follows: arterial blood gas procedures, 92.3%; intravenous line initiation/maintenance, 77.6%; and phlebotomy, 70.6%. Glove use during phlebotomy (p less than 0.001) and intravenous line procedures (p less than 0.05) was significantly lower in the state with a prevalence of the acquired immunodeficiency syndrome (AIDS) below the national average than in the states with a higher AIDS prevalence. The investigation suggests that health care facilities have responded to the Centers for Disease Control and Occupational Safety and Health Administration campaign to adopt universal precaution policies for glove use by HCWs. Actual glove use by HCWs appears to be substantial but not universal. Glove use by HCWs is significantly related to statewide AIDS prevalence.

Acquired Immunodeficiency Syndrome

T-lymphocyte requirement for diabetes in RT6-depleted diabetes-resistant BB rats.

Diabetes-prone (DP) BB rats develop spontaneous autoimmune insulin-dependent diabetes mellitus (IDDM). The cell populations involved in the expression of diabetes are not precisely known but probably include natural killer (NK) cells, macrophages, and T lymphocytes. Because the DP rat has few lymphocytes of the CD5+/CD+ phenotype, cytotoxic T lymphocytes (Tc) are not believed to be important in the process. Diabetes-resistant (DR) BB rats that are depleted of RT6+ T lymphocytes also become diabetic and provide an additional model of IDDM. We report that diabetes in DR rats depleted of RT6+ T lymphocytes is prevented by the concomitant depletion of either the CD5+ or the CD8+ population. In contrast, coadministration of anti-asialogangliosideM1 (alpha-ASGM1), an antiserum that principally recognizes NK cells, failed to prevent hyperglycemia in RT6-depleted rats. We propose that the initiation of diabetes in both DP and RT6-depleted DR rats is T-lymphocyte dependent. However, the final common pathway leading to autoimmune beta-cell destruction in IDDM may be different in these models. The RT6-depleted DR rat requires a cell that is sensitive to anti-CD8 (possibly a Tc), whereas the DP rat requires an anti-ASGM1-sensitive cell.

ADP Ribose Transferases

Vocal function following vertical hemilaryngectomy: a preliminary investigation.

Seven consecutive male patients who received a vertical hemilaryngectomy were examined for vocal function employing video-stroboscopic, aerodynamic and acoustic measurement techniques. The results of this preliminary investigation suggest that with the group of patients studied, vocal quality was rated as "rough", "breathy" and "constricted". While large variability was noted between individuals, the general tendencies for the group included: (a) incomplete glottic closure; (b) supraglottic structures (ventricular folds, arytenoids) fulfilled or aided vibratory action of the remaining fold; (c) high average transglottal airflow; (d) reduced maximum phonation time; (e) high and more variable vocal frequency positioned near the top of the frequency range; (f) a lower, more variable, and restricted vocal intensity range, and (g) a reduced vocal fold diadochokinetic (vocal fold opening and closing action) rate. We are currently attempting to determine factors related to individual patient strategies for using either glottic or supraglottic valving during phonation.

Carcinoma, Squamous Cell

The olfactory and cognitive deficits of Parkinson's disease: evidence for independence.

In the present study we sought to determine whether the olfactory deficits of parkinsonism are related to the cognitive and perceptual-motor manifestations of the disease. Pearson correlation coefficients were computed among a number of olfactory, neurological, and neuropsychological measures obtained from 58 Parkinson's disease patients, including the University of Pennsylvania Smell Identification Test, a modified Randt memory test, a reaction time test, a finger-tapping test, ratings of motor and neurological function, and selected verbal and performance subtests of the Wechsler Adult Intelligence Scale--Revised. Data from 38 patients with Parkinson's disease who received odor detection threshold testing were also evaluated. The intercorrelation matrix was subjected to a principal components factor analysis which yielded six clear-cut factors: cognitive/memory, gross motor, oral motor, fine motor, olfactory, and tremor. The olfactory factor received strong and nearly exclusive loadings from the olfactory measures (which did not evidence meaningful loadings on any of the other factors). A ten-trial jackknife procedure revealed the factor structure to be stable. Further support of the independence of the olfactory variables from the other measures was provided by multiple regression and canonical correlation analyses. Overall, these findings lend support to the hypothesis that the olfactory disorder of parkinsonism is independent of the cognitive, perceptual-motor, and memory manifestations of the disease.

Aged

Spatial dispersion of stainable lipid in frozen sections of human aorta.

Frozen sections of human lateral thoracic aorta were stained for lipid with Oil red O. Intensity of staining was judged upon a scale of 0 to 5 brilliance units by matching to a scale of photographs. Microregions, defined by an eyepiece grid, were structured in layers of 100 microns thickness and 1000 microns along the aortic length. The intensity of staining, both total and extracellular, tended significantly to increase with age, intimal thickness and depth into the intima. Aortas having at least one instance of atheronecrosis tended to show generally greater staining intensities than aortas without necrosis, and the tendency was more conspicuous with total than with extracellular staining, implying a greater degree of intracellular staining in the specimens with atheronecrosis. The evidence seems to suggest that the emergence of a necrotic core happens, at least in part, because the affected site has collected an excess of lipid. The evidence further suggests that lipid deposits become excessive with increasing frequency after age 35 years, and that this is associated with intimal thickenings coming to exceed 300 microns. The causes that propel the intimal thickenings remain wholly unknown - those causes seem to have little if anything to do with excess of lipids. Some as yet undetermined qualitative characteristic of the lipids might be of importance in this regard.

Adolescent