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Biomedical subjects

D Fuller

Publications and source records attributed to D Fuller.

At least 37 records · Page 2Linked to original sources

DNA vaccines expressing either the GP or NP genes of Ebola virus protect mice from lethal challenge.

DNA vaccines expressing the envelope glycoprotein (GP) or nucleocapsid protein (NP) genes of Ebola virus were evaluated in adult, immunocompetent mice. The vaccines were delivered into the skin by particle bombardment of DNA-coated gold beads with the Powderject-XR gene gun. Both vaccines elicited antibody responses as measured by ELISA and elicited cytotoxic T cell responses as measured by chromium release assays. From one to four vaccinations with 0.5 microgram of the GP DNA vaccine resulted in a dose-dependent protection from Ebola virus challenge. Maximal protection (78% survival) was achieved after four vaccinations. Mice were completely protected with a priming dose of 0.5 microgram of GP DNA followed by three or four subsequent vaccinations with 1.5 micrograms of DNA. Partial protection could be observed for at least 9 months after three immunizations with 0.5 microgram of the GP DNA vaccine. Comparing the GP and NP vaccines indicated that approximately the same level of protection could be achieved with either vaccine.

Animals↗

Co-activation of tongue protrudor and retractor muscles during chemoreceptor stimulation in the rat.

1. Our primary purpose was to test the hypothesis that the tongue protrudor (genioglossus, GG) and retractor (styloglossus, SG and hyoglossus, HG) muscles are co-activated when respiratory drive increases, and that co-activation will cause retraction of the tongue. This was addressed by performing two series of experiments using a supine, anaesthetized, tracheotomized rat in which tongue muscle force and the neural drive to the protrudor and retractor muscles could be measured during spontaneous breathing. In the first series of experiments, respiratory drive was increased progressively by occluding the tracheal cannula for thirty respiratory cycles; in the second series of experiments, the animals were subjected to hyperoxic hypercapnia and poikilocapnic hypoxia. 2. Airway occlusion for thirty breaths caused progressive, quantitatively similar increases in efferent motor nerve activity to protrudor and retractor tongue muscles. Net tongue muscle force was always consistent with tongue retraction during occlusion, and peak force rose in parallel with the neural activites. When airway occlusion was repeated following section of the lateral XIIth nerve branch (denervation of retractor muscles) the tongue either protruded (15/21 animals; 10 +/- 2 mN at the 30th occluded breath) or retracted weakly (6/21 animals; 6 +/- 2 mN at 30th occluded breath). 3. To ensure that our findings were not the result of damage to the muscle nerves, occlusion experiments were also done in eight animals in which GG EMG activity was recorded instead of nerve activities. Changes in peak integrated GG electryomyogram (EMG) activity and peak retraction force during occlusion were highly correlated (r2 = 0.86, slope = 1.05). 4. In separate experiments in fourteen rats, we found that hyperoxic hypercapnia and poikilocapnic hypoxia also result in parallel increases in the respiratory-related EMG activity of the GG and HG muscles. Also, as in the occlusion experiments, augmentations of protrudor and retractor muscle EMG activities were associated with parallel changes in tongue retraction force. 5. These studies in anaesthetized rats demonstrate that tracheal occlusion and independent stimulation of central or peripheral chemoreceptors results in inspiratory-related co-activation of the protrudor and retractor muscles, and proportional changes in tongue retraction force. These observations also demonstrate that recording GG EMG activity in isolation could lead to erroneous conclusions about respiratory-related movements of the tongue.

Airway Obstruction↗

Do patients refusing transport remember descriptions of risks after initial advanced life support assessment?

OBJECTIVE: To determine patient recall and understanding of instructions given to patients who refuse transport after initial paramedic assessment and medical treatment. METHODS: Following patient consent, a phone interview was completed for consecutive persons living in a large urban area for whom 9-1-1 was contacted but who subsequently refused transport after advanced life support (ALS) assessment. Subjects were asked about their recall of explained risks and benefits of transport, their understanding of those risks at the time of assessment, and subsequent use of medical care, including hospitalization. RESULTS: From October 1, 1996, to February 23, 1997, 324 people refused transport after ALS arrival. Sixty-eight people could not be contacted, providing a response rate of 79% (256/324). Six percent were subsequently admitted to the hospital for the same problem and an additional 59% sought care from a health care provider (66 ED visits, 63 personal physician, 16 urgent care, 5 other). There were no unexpected deaths. Ninety (35%) respondents were still experiencing symptoms at the time of phone contact. Despite the routine practice of providing a verbal explanation of risks and written instructions, only 141 (55%) recalled receiving written instructions and 56 (22%) recalled an explanation of risks. Twenty-six percent believed they did not fully understand their conditions or circumstances surrounding the 9-1-1 call when they refused transport and 18% would now take an ambulance if the same incident were to recur. CONCLUSION: A substantial proportion of patients refusing transport do not recall receiving verbal or written instructions and would reconsider their transport decision, raising doubts about people's ability to make informed decisions at a time of great vulnerability. The majority of patients accessed health care after refusing transport and 6% were hospitalized.

Emergency Medical Services↗

Multivariate analysis of objective vocal function.

No standard and valid multidimensional index of objective voice function has been developed that integrates the information generated from the multiple objective parameters of voice function. The goals of this research were 1) to identify important objective voice parameters and 2) to create a multidimensional voice function index by combining relevant parameters. We evaluated 97 dysphonic patients and 35 normal volunteers on 14 objective voice parameters. Three multidimensional voice indices were created and evaluated: 1) nonweighted univariate index, 2) weighted odds ratio index, and 3) weighted multivariate regression index. The univariate index required all 14 parameters, while the odds ratio and logistic regression models required only 4 parameters (frequency range, airflow at lips, maximum phonation time, and subglottic pressure). The chi2 values for the 3 models were 37.8, 37.6, and 46.0, respectively. All 3 indices were able to satisfactorily classify voice function as normal or abnormal. However, the regression index performed best.

Confidence Intervals↗

Assessment of two objective voice function indices.

In the care of patients with voice disorders, physicians, speech pathologists, and other health care professionals routinely make diagnoses, recommend treatment, and evaluate outcomes. Although objective and subjective measures exist, unfortunately, there is no widely accepted, valid method for classifying voice disorders and assessing outcome after voice treatment. In the present research, the relationship between two previously created multivariate objective voice function indices, the weighted odds ratio index and the multivariate logistic regression index, and subjective assessment of voice function was evaluated. Twenty-three adult patients presenting to a speech science laboratory for evaluation of voice disorders were studied in this prospective observational study together with 12 normal volunteers as controls. Vocal function was measured on 14 different parameters with a protocol that included a multichannel input for simultaneous assessment of acoustic and physiological parameters. Each patient was recorded reading the standard passage "The North Wind and the Sun," and recordings were then evaluated by the GRBAS scale. Overall, there was a statistically significant relationship between the weighted odds ratio index and multivariate logistic regression index and mean GRBAS scores. This research demonstrates that the voice function values calculated from two different multivariate objective voice function indices are significantly associated with subjective voice assessments. These multivariate objective voice indices may be appropriate for use in clinical trials and outcomes research on treatment effectiveness for voice disorders.

Adult↗

A cAMP-phosphodiesterase controls PKA-dependent differentiation.

A cAMP-specific phosphodiesterase was found that is stimulated by binding to the regulatory subunit of cAMP-dependent protein kinase, PKA-R, from either Dictyostelium or mammals. The phosphodiesterase is encoded by the regA gene of Dictyostelium, which was recovered in a mutant screen for strains that sporulate in the absence of signals from prestalk cells. The sequence of RegA predicts that it will function as a member of a two-component system. Genetic analyses indicate that inhibition of the phosphodiesterase results in an increase in the activity of PKA, which acts at a check point for terminal differentiation. Conserved components known to affect memory, learning and differentiation in flies and vertebrates suggest that a similar circuitry functions in higher eukaryotes.

3',5'-Cyclic-AMP Phosphodiesterases↗

Consequences of disrupting the gene that encodes alpha-glucosidase II in the N-linked oligosaccharide biosynthesis pathway of Dictyostelium discoideum.

We have identified and disrupted the gene coding for alpha-glucosidase II in Dictyostelium discoideum. This enzyme is responsible for removing two alpha 1,3-linked glucose residues from N-linked oligosaccharides on newly synthesized glycoproteins. Mutagenesis by restriction enzyme-mediated integration (REMI) generated a clone, DG1033, which grows well but forms abnormal fruiting bodies with short, thick stalks. The strain lacks alpha-glucosidase II activity and makes incompletely processed N-linked oligosaccharides that are abnormally large and have fewer sulfate and phosphate esters. The morphological, enzymatic, and oligosaccharide profile phenotypes of the disruption mutant are all recapitulated by a targeted disruption of the normal gene. Furthermore, all of these defects are corrected in cells transformed with a normal, full-length copy of the gene. The phenotypic characteristics of DG1033 as well as chromosomal mapping of the disrupted gene indicate that it is the site of the previously characterized modA mutation. The Dictyostelium gene is highly homologous to alpha-glucosidase II genes in the human and the pig, C. elegans, and yeast. Although various cell lines have been reported to be defective in alpha-glucosidase II activity, disruption of the Dictyostelium gene gives the first example of a clear developmental phenotype associated with loss of this enzyme.

Amino Acid Sequence↗

Naked DNA vaccines expressing the prM and E genes of Russian spring summer encephalitis virus and Central European encephalitis virus protect mice from homologous and heterologous challenge.

Naked DNA vaccines expressing the prM and E genes of two tick-borne flaviviruses, Russian spring summer encephalitis (RSSE) virus and Central European encephalitis (CEE) virus were evaluated in mice. The vaccines were administered by particle bombardment of DNA-coated gold beads by Accell gene gun inoculation. Two immunizations of 0.5 to 1 microg of RSSE or CEE constructs/dose, delivered at 4-week intervals, elicited cross-reactive antibodies detectable by enzyme-linked immunosorbent assay and high-titer neutralizing antibodies to CEE virus. Cross-challenge experiments demonstrated that either vaccine induced protective immunity to homologous or heterologous RSSE or CEE virus challenge. The absence of antibody titer increases after challenge and the presence of antibodies to E and prM, but not NS1, both before and after challenge suggest that the vaccines prevented productive replication of the challenge virus. One vaccination with 0.5 microg of CEE virus DNA provided protective immunity for at least 2 months, and two vaccinations protected mice from challenge with CEE virus for at least 6 months.

Animals↗

Critical thinking in undergraduate athletic training education.

OBJECTIVE: The purposes of this study were (a) to determine whether or not undergraduate athletic training educators are writing learning objectives that foster critical thinking (CT) skills, and (b) to determine if their written assignments and written examinations are measuring the extent to which students have developed CT skills. DESIGN AND SETTING: Thirty institutions seeking accreditation for their athletic training programs from the Commission on Accreditation of Allied Health Educational Programs in the 1994-95 academic year were asked to provide their curriculum materials (course syllabus, two to three examinations, or both from each athletic training-specific course). SUBJECTS: Thirteen curriculum directors (43%) provided materials. MEASUREMENTS: Each learning objective, examination question, and written assignment was classified as either CT or non-critical thinking (NCT) using Bloom's taxonomy. RESULTS: From 64 usable syllabi, a total of 678 learning objectives were classified as either CT (52%) or NCT (48%). From 81 written examinations, 3215 questions were classified as either CT (14%) or NCT (86%). In addition, a total of 143 written assignments were all classified as CT. CONCLUSIONS: The results of this study indicate that educators fostered more CT in their learning objectives and written assignments than in their written exams. Valid educational instruments (eg, Bloom's taxonomy) may help educators design learning objectives, assignments, and examinations.

Journal Article↗

Cell-cell adhesion prevents mutant cells lacking myosin II from penetrating aggregation streams of Dictyostelium.

When a small number of fluorescently labeled myosin II mutant cells (mhcA-) are mixed with wild-type cells and development of the chimeras is observed by confocal microscopy, the mutant cells are localized to the edges of aggregation streams and mounds. Moreover, the mutant cells stick to wild-type cells and become distorted (Shelden and Knecht, 1995). Two independent adhesion mechanisms, Contact Sites A and Contact Sites B, function during the aggregation stage and either one or both might be responsible for excluding the myosin II null cells. We have mixed mhcA- cells with cells in which the appearance of Contact Sites B is delayed (strain TL72) as well as cells which lack Contact Sites A (strain GT10) and double mutants in which both adhesion mechanisms are affected (strain TL73). In all chimeras, the mhcA- cells were distorted by interactions with the adhesion mutant cells, indicating that it does not require significant adhesive interaction to distort the flaccid cortex of mhcA- cells mhcA- cells were excluded from streams composed of cells lacking either Contact Sites A or Contact Sites B but mixed randomly with cells lacking both adhesion systems. By 10 hr of development, cells of strain TL73 acquire Contact Sites B adhesion. If cells of this strain were mixed with labeled mhcA- cells, allowed to develop for 9 hr, and then dissociated before replating, the myosin II null cells were seen to be distorted and excluded from the reaggregates. Thus, the exclusion of mhcA- cells from streams can be accomplished by either Contact Sites A or B. When chimeras of labeled TL73 and wild-type cells were made, the TL73 cells were found to be randomly mixed into aggregation streams. This result indicates that adhesive sorting does not function during aggregation and so cannot account for the exclusion of mhcA- cells from streams. We hypothesize that the flaccid cortex of mhcA- cells cannot generate sufficient protrusive force to break the contacts between adhered cells in aggregation streams but can enter streams where the cells are weakly adherent.

Amino Acid Sequence↗

Expiratory muscle endurance performance after exhaustive submaximal exercise.

The aim of our study was to describe the endurance capacity of the expiratory muscles and to determine whether it is altered after exhaustive cycling exercise. Subjects performed repeated maximal expiratory efforts against a closed breathing valve, with and without prior exercise performed at a work rate that elicited 75% of the maximum ventilation rate. Each expiratory effort lasted 6 s, was separated by 10 s of rest, and was initiated from the end-expiratory lung volume. Endurance performance was assessed by measuring the decline in area under the pressure*time curve over 39 contractions. Prior exhaustive exercise attenuated the ability to generate and sustain maximal expiratory pressure (P = 0.013) and resulted in significant declines in the integrated electromyogram of the rectus abdominis (P = 0.005) and external oblique (P = 0.036) abdominal muscles. Each subject also performed a handgrip endurance task before and after exhaustive exercise on a separate day. Prior exercise had no effect on handgrip endurance performance, suggesting that the decline in expiratory muscle performance after exercise was not the result of reduced motivation. We conclude that the ability to maximally activate the abdominal expiratory muscles and to generate maximum expiratory pressure is impaired after exhaustive exercise. Declines in the surface integrated electromyogram despite maximal effort is consistent with findings in limb muscles and is thought to be due to a slowing of motoneuron firing rates or to neuromuscular transmission failure.

Adult↗

Control of nasal dilator muscle activities during exercise: role of nasopharyngeal afferents.

Our primary aim was to determine whether reducing the activity of nasal airway receptors would influence drive to the nasal dilator muscles (NDMs) during exercise. We used lidocaine (2%) or nasal splints to diminish afferent airway receptor activity and measured the electromyogram (EMG) activity of the NDMs during incremental bicycle exercise in subjects who breathed nasally. NDM EMG activities increased as a function of exercise intensity but were not changed by lidocaine and were only slightly reduced by splinting. Similarly, neither intervention altered the normal decrease in NDM EMG activity associated with reductions in airway resistance evoked by He-O2 breathing. We also compared the NDM EMG response to exercise with that evoked by CO2 rebreathing at rest to determine whether the nature of the ventilatory stimulus influences drive to the NDMs; comparisons were made at constant levels of nasal inspired ventilation and, therefore, constant total ventilatory output. The increase in EMG activity was much higher during exercise compared with hyperoxic hypercapnia. In conclusion, 1) desensitizing the nasal airway does not alter NDM activity significantly during exercise and 2) exercise results in much greater increases in NDM activity compared with hypercapnia, indicating that different ventilatory stimuli can evoke more or less activation of upper airway motoneurons, even when comparisons are made at constant levels of total ventilatory output.

Adult↗

Measurement of the EMG-force relationship in a human upper airway muscle.

The upper airway muscles play an important role in breathing, swallowing, and speaking, but little is known about the electromyogram (EMG)-force relationship of these muscles. We have measured the peak integrated EMG activity (iEMG) and force of human nasal dilator muscles (NDM) with a custom-designed headpiece that was attached via the forehead and upper lip. The headpiece contains a micromanipulator that holds a rod with a load cell mounted on its tip. The reproducibility of the force measurements was examined by measuring the lateral or "flaring" force of the NDM in multiple trials on two separate occasions in 13 subjects. For these studies the subjects were instructed to perform maximal voluntary contractions (MVCs). Test-retest reproducibility averaged 8.3% (coefficient of variation) for within-day comparisons and 13.7% between days. We also measured iEMG and NDM force during an incremental exercise test in nine of the subjects; they were instructed to breathe nasally throughout one 30-s epoch at rest and at each workload. The iEMG and force during peak exercise (175-275 W) averaged 81 +/- 26% (SD) MVC and 235 +/- 127 mN (approximately 75% MVC), respectively. The iEMG during incremental exercise was linearly related to the peak force (r = 0.90, P < 0.001). Contractile properties were measured in seven of the subjects by application of single supramaximal shocks (0.1-ms pulse) to the facial nerve. Twitch force averaged 9 +/- 6% MVC, and the time to peak force was 62 +/- 13 ms, which is considerably faster than that in human diaphragm or elbow flexors.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Vibratory segment function after free flap reconstruction of the pharyngoesophagus.

Reconstructive options following total laryngopharyngectomy include thin, pliable free tissue segments, approximating the natural thickness of the pharyngeal wall. The authors have investigated outcomes in the following clinical series, emphasizing speech and swallowing. Twelve cancer patients underwent laryngopharyngectomy with or without glossectomy. Eight jejunal, 1 radial forearm, and 3 innervated latissimus dorsi flaps were used for vibratory segment (VS) reconstruction, and all 12 patients underwent tracheoesophageal puncture (TEP). Eleven patients achieved intelligible speech, with a median intelligibility of 93%. The vibrating segments showed fluttering of the free flap tissue when studied by videopharyngography. Vocal quality was lower pitched and softer than "conventional" TEP speech. All patients achieved oral intake as their primary mode of nutrition. Free flaps are a successful option for VS reconstruction in patients undergoing laryngopharyngectomy or glossopharyngolaryngectomy, obviating the need for written or electrolarynx communication.

Aged↗

Simplifying the system. Assessing drug administration methods.

1. Discontinuing the medicine round is an important step towards providing truly individualised patient care. 2. Personalised drug dispensing systems promote effective use of nursing time. 3. Hospital-run self-administration of drugs programmes can provide assessment knowledge with which to plan patients' future drug administration needs.

Humans↗

Structural roles of the spore coat proteins in Dictyostelium discoideum.

The integrity of spores formed by mutant strains of Dictyostelium discoideum lacking the major spore coat proteins, SP96, SP70, or SP60, was compared to that of wild-type strains. Single, double, and triple knock-out strains developed normally and produced spores which were indistinguishable from wild-type spores by light or electron microscopy. However, the mutant strains were susceptable to staining with the lectin, ricin A, which recognizes a galactose-rich polysaccharide that is normally hidden by overlying spore coat proteins. The intensity of staining with fluorescently labeled ricinA increased as the spore coat proteins were incrementally lost. While these results indicate that the major outer spore coat proteins are not essential for the construction of a multi-layered spore coat in Dictyostelium, they show that the spores are more porous which might make them at risk to predators before germination.

Animals↗

Measurement of felbamate by wide-bore capillary gas chromatography and flame ionization detection.

Felbamate, a newly developed antiepileptic agent, has been demonstrated to control partial and generalized seizures effectively. We have developed a gas-chromatographic method for the determination of felbamate, using a wide-bore capillary column, a flame ionization detector, and a simple extraction procedure. The assay day-to-day precision (n = 20) was 5.2% and 3.6% for drug concentrations of 50 and 150 mg/L, respectively; average recovery over a wide range of felbamate concentrations was 95%; the detection limit was 5 mg/L; and assay linearity extended to 300 mg/L. Although 9 of the 27 drugs tested were coextracted with felbamate, they exhibited significantly different retention times and showed no interference. A short-term stability study showed that plasma felbamate is stable at 4, -20, or -78 degrees C for at least 1 month. Plasma felbamate concentrations in 66 pediatric patients ranged from 7 to 154 mg/L (mean +/- SD 44 +/- 24.7). We consider the method ideally suited for therapeutic monitoring of plasma felbamate concentration.

Anticoagulants↗