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Biomedical subjects

C E Morris

Publications and source records attributed to C E Morris.

At least 37 records · Page 2Linked to original sources

Transepithelial transport of nicotine and vinblastine in isolated malpighian tubules of the tobacco hornworm (Manduca sexta) suggests a P-glycoprotein-like mechanism.

We have examined the accumulative transport properties of the Malpighian (excretory) tubules of the tobacco hornworm Manduca sexta to test the hypothesis that a P-glycoprotein-like multidrug transporter is active and is responsible for the excretion of dietary nicotine in this tissue. Isolated tubules were cannulated and exposed to radiolabelled forms of either nicotine (5 min exposure) or the P-glycoprotein substrate vinblastine (60 min exposure) in the bathing (basal surface) fluid. The luminal (apical) contents were then flushed, and lumen-to-bath ratios were measured. Although these ratios provide conservative estimates of the physiological ability of Malpighian tubules to move compounds from blood to lumen, tubules concentrated nicotine 10-fold from an initial bath concentration of 0.5 mmol l-1 and vinblastine threefold (from an initial concentration of 1 micromol l-1). Vectorial transport of vinblastine and nicotine was eliminated by 25 micromol l-1 verapamil (a P-glycoprotein inhibitor) and was not dependent on the presence of a transepithelial electrical potential. Nicotine transport was inhibited by atropine (3 mmol l-1), while nicotine (> or = 50 micromol l-1) significantly reduced vinblastine transport. Verapamil was effective at reducing vinblastine transport when applied to the basal side alone, but not when applied to the apical side alone. Taken together, these results are consistent with the idea that the active excretion of nicotine and other alkaloids by the tobacco hornworm is mediated by a P-glycoprotein-like mechanism.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Biocompatibility of sweetpotato and peanut in a hydroponic system.

'Georgia Red' peanut (Arachis hypogaea L.) and TU-82-155 sweetpotato [Ipomoea batatas (L.) Lam] were grown in monocultured or intercropped recirculating hydroponic systems in a greenhouse using the nutrient film technique (NFT). The objective was to determine whether growth and subsequent yield would be affected by intercropping. Treatments were sweetpotato monoculture (SP), peanut monoculture (PN), and sweetpotato and peanut grown in separate NFT channels but sharing a common nutrient solution (SP-PN). Greenhouse conditions ranged from 24 to 33 degrees C, 60% to 90% relative humidity (RH), and photosynthetic photon flux (PPF) of 200 to 1700 micromoles m-2 s-1. Sweetpotato cuttings (15 cm long) and 14-day-old seedlings of peanuts were planted into growth channels (0.15 x 0.15 x 1.2 m). Plants were spaced 25 cm apart within and 25 cm apart between growing channels. A modified half-Hoagland solution with a 1 N: 2.4 K ratio was used. Solution pH was maintained between 5.5 and 6.0 for treatments involving SP and 6.4 and 6.7 for PN. Electrical conductivity (EC) ranged between 1100 and 1200 microS cm-1. The number of storage roots per sweetpotato plant was similar for both SP and SP-PN. Storage root fresh and dry mass were 29% and 36% greater, respectively, for plants in the SP-PN treatment than for plants in the SP treatment. The percent dry mass of the storage roots, dry mass of fibrous and pencil roots, and the length-to-diameter ratio of storage roots were similar for SP and SP-PN sweetpotato plants. Likewise, foliage fresh and dry mass and harvest index were not significantly influenced by treatment. Total dry mass was 37% greater for PN than for SP-PN peanut plants, and pod dry mass was 82% higher. Mature and total seed dry mass and fibrous root dry mass were significantly greater for PN than for SP-PN plants. Harvest index (HI) was similar for both treatments. Root length tended to be lower for seedlings grown in the nutrient solution from the SP-PN treatment.

Arachis↗

Initial and steady-state effects of diphenhydramine and loratadine on sedation, cognition, mood, and psychomotor performance.

BACKGROUND: The classic, first-generation histamine1-receptor antagonists used to treat allergic disorders frequently cause sedation. In contrast, sedation is reduced or absent after administration of recommended doses of second-generation histamine1-receptor antagonists. We measured the initial and steady-state effects of diphenhydramine, a first-generation antihistamine, and loratadine, a second-generation antihistamine, by means of a comprehensive battery of psychometric tests that mirror real-world tasks. METHODS: Healthy volunteers (N = 98) were randomly assigned in a double-blind fashion to receive loratadine (n = 33), diphenhydramine (n = 32), or placebo (n = 33). A computerized test battery was administered at baseline, on day 1 after administration of the initial dose, and on days 3 and 5. RESULTS: After the initial dose, subjects taking diphenhydramine demonstrated poorer cognitive performance than subjects taking loratadine or placebo on tasks of divided attention, working memory, speed, and vigilance. Subjects taking diphenhydramine also reported greater fatigue and sleepiness and lower levels of motivation, and rated the quality of their performance as lower than subjects taking loratadine or placebo. On day 3, subjects taking diphenhydramine continued to show more fatigue and lower motivation, and rated the quality of their test performance as poorer than subjects taking loratadine or placebo. There were no differences between loratadine and placebo after the initial dose or steady-state (day 5) dosing for any measure of cognitive or psychomotor test performance, mood, or sedation. CONCLUSIONS: Patients taking diphenhydramine may be at risk of lapses and significant errors that may lead to potential hazards and decreased work productivity.

Adult↗

Methods for observing microbial biofilms directly on leaf surfaces and recovering them for isolation of culturable microorganisms.

Epifluorescence microscopy, scanning electron microscopy, and confocal laser scanning microscopy were used to observe microbial biofilms directly on leaf surfaces. Biofilms were observed on leaves of all species sampled (spinach, lettuce, Chinese cabbage, celery, leeks, basil, parsley, and broad-leaved endive), although the epifluorescent images were clearest when pale green tissue or cuticle pieces were used. With these techniques, biofilms were observed that were about 20 (mu)m in depth and up to 1 mm in length and that contained copious exopolymeric matrices, diverse morphotypes of microorganisms, and debris. The epifluorescence techniques described here can be used to rapidly determine the abundance and localization of biofilms on leaves. An additional technique was developed to recover individual biofilms or portions of single biofilms from leaves and to disintegrate them for isolation of the culturable microorganisms they contained. Nineteen biofilms from broad-leaved endive, spinach, parsley, and olive leaves were thus isolated and characterized to illustrate the applications of this technique.

Journal Article↗

Human subject research at Armstrong Laboratory, 1973-93: medical and musculoskeletal disqualifications.

The reasons for disqualification of human subjects from 1973-93 at Armstrong Laboratory, formerly the Harry G. Armstrong Aeromedical Research Laboratory (AAMRL), are presented for both sustained and impact acceleration panels. Evaluations for both medical and spinal anomalies were accomplished. The rationale for each disqualification is discussed, demonstrating the variability in each panel physician's clinical judgment in the context of personal bias, the ethical framework surrounding the use of human volunteers, and the existing research milieu.

Acceleration↗

Comparison of acceleration subjects to other populations: spinal anomaly distribution.

Two Armstrong Laboratory (AL) human volunteer subject panels (sustained and impact acceleration) at Wright Patterson AFB, OH, were compared to each other and to other samples of different populations in terms of spinal anomalies. These sample populations were obtained from the scientific literature: French, Norwegian, Netherlands, and U.S. pilots; U.S. Air Force (AF) and Navy subjects, and from representative "normal" civilian populations, and then compared using the proportion parameters for various spinal anomalies. There were only a few common parameters between the two panels and between each panel compared with the foreign military, human subjects, and "normal" population. However, there were two to six times as many similar spinal anomaly incidence rates between the AL panels and the U.S. pilot sample. It was reassuring that the AL subject panels used in AF acceleration research have more in common with AF pilots than other populations in regards to spinal anomalies, even though the pilot sample may not be representative of the true pilot population. Recommendations are to establish a common reference point in nomenclature and description of spinal anomalies (modeled after the French) and to start collecting spinal radiographs on all U.S. pilots. These radiographs would not be for screening but for establishing a database following the occupational pathology of flying. This data would also facilitate comparisons with research acceleration panels, as well as with foreign air forces. Informed decisions can then be made regarding screening criteria for the future as aircraft and ejection seat performance envelopes continue to expand.

Acceleration↗

Are human subject volunteers still players in aeromedical research as we enter the 21st century?

The U.S. Air Force has enjoyed the luxury of having dedicated human volunteer subjects for sustained and impact acceleration research for over 50 yr. However, with today's world economy and budgetary cutbacks, this may no longer be a viable option. The onslaught of advanced medical technology, combined with an increasing performance envelope for aircraft and their ejection systems, have created an environment where the validity of research data and the ethics of human-use research are being challenged. Now is an opportune time to reevaluate the way human-use aeromedical research is conducted. The validity of using nonpilots in lieu of pilots in aeromedical research is discussed in light of the following: a) the increased emphasis on performance metrics within sustained acceleration; b) the matching of human subjects (nonpilots) to pilots in the appropriate attributes to ensure validity of data; c) degree of medical screening required given the ethics of human-use research and concerns of pilots; and d) the challenge of evaluating the "value added" of new technology for medical screening. It is concluded that volunteer panels should be maintained with nonpilots matched with pilots physically and psychologically such that operational performance characteristics are similar. Medical screening should be similar so that research data from subjects can be applied to the target population (pilots). Longitudinal data collection (e.g., spinal X-rays) on pilots would also be of great value as a basis for studying the occupational hazards of flying.

Acceleration↗

Human subject screening: a dynamic process.

INTRODUCTION: The history of disqualified (DQ) subjects from 1973-1993 at Armstrong Laboratory, Wright Patterson AFB, is presented for both sustained and impact acceleration panels. METHODS: Candidate and subject medical records were reviewed for screening results, recommendation for panel duty, and any follow-up medical findings. The generation and interpretation of the medical screening criteria and DQ rates are discussed. MEDICAL SCREENING CRITERIA: The mechanisms for change, those factors influencing change, and the interpretation of the screening criteria for Armstrong Laboratory's acceleration panels determine the panel's composition, which is reflected in the DQ rates. RESULTS: The centrifuge had a 5% (7/132) disqualification (DQ) rate from 1973-93 with 29% (2/7) due to musculoskeletal and 71% (5/7) for medical reasons. All were DQ during 1973-88. The impact panel had a DQ rate of 18% (36/195) with 71% (24/34) DQ due to musculoskeletal and 29% (10/34) for medical reasons. Only 28% (10/36) were DQ during 1973-88, while during 1989-93, 72% (26/36) were DQ. CONCLUSIONS: The differences in DQ rates between the centrifuge and impact facility were due to the variability or conservatism of individual physicians, interpretation of the medical screening criteria, and the type of research being done. These factors effect the composition of the human subject panels. This determines to which target population the research data can be applied. If the subjects do not represent pilots due to inappropriate screening, then there is no benefit from the research and, therefore, there can be no risk incurred by the subjects.

Acceleration↗

Coiled mechanoreceptors in Aplysia revealed by sensorin immunofluorescence and confocal microscopy.

Identified mechanosensory neurons of Aplysia are established model neurons for studies on learning and memory, and for examining responses to axonal injury. Although many characteristics of these sensory neurons have received intensive study, the nature of the peripheral mechanoreceptive endings remains unknown. Identification of a peptide, sensorin, specific in Aplysia for mechanosensory neurons, led to the development of an antibody which proved useful in studying the peripheral morphology of these neurons. Immunostaining for sensorin in tail body wall revealed that sensorin is present in peripheral arborizations. Examination of sensorin-positive fibers in the periphery revealed that they terminate as coiled structures in the muscle layer of the body wall. These coiled structures (approximately 0.5 microns diameter processes, 2-3 microns across the coil, approximately 60 microns long) run parallel to muscle fibers and have a pitch of about one turn per 4 microns. Sensorin immunostaining was particularly intense in varicosities, both along peripheral fibers and along the coiled structure. The localization of sensorin suggests that it may be released peripherally where it could have various paracrine and/or autocrine neuromodulatory actions.

Animals↗

Responses of neurons to extreme osmomechanical stress.

Neurons are often regarded as fragile cells, easily destroyed by mechanical and osmotic insult. The results presented here demonstrate that this perception needs revision. Using extreme osmotic swelling, we show that molluscan neurons are astonishingly robust. In distilled water, a heterogeneous population of Lymnaea stagnalis CNS neurons swelled to several times their initial volume, yet had a ST50 (survival time for 50% of cells) > 60 min. Cells that were initially bigger survived longer. On return to normal medium, survivors were able, over the next 24 hr, to rearborize. Reversible membrane capacitance changes corresponding to about 0.7 muF/cm2 of apparent surface area accompanied neuronal swelling and shrinking in hypo- and hyperosmotic solutions; reversible changes in cell surface area evidently contributed to the neurons' ability to accommodate hydrostatic pressures then recover. The reversible membrane area/capacitance changes were not dependent on extracellular Ca2+. Neurons were monitored for potassium currents during direct mechanical inflation and during osmotically driven inflation. The latter but not the former stimulus routinely elicited small potassium currents, suggesting that tension increases activate the currents only if additional disruption of the cortex has occurred. Under stress in distilled water, a third of the neurons displayed a quite unexpected behavior: prolonged writhing of peripheral regions of the soma. This suggested that a plasma membrane-linked contractile machinery (presumably actomyosin) might contribute to the neurons' mechano-osmotic robustness by restricting water influx. Consistent with this possibility, 1 mM N-ethyl-maleimide, which inhibits myosin ATPase, decreased the ST50 to 18 min, rendered the survival time independent of initial size, and abolished writhing activity. For neurons, active mechanical resistance of the submembranous cortex, along with the mechanical compliance supplied by insertion or eversion of membrane stores may account for the ability to withstand diverse mechanical stresses. Mechanical robustness such as that displayed here could be an asset during neuronal outgrowth or regeneration.

Animals↗

Discrete and reversible vacuole-like dilations induced by osmomechanical perturbation of neurons.

In cultured Lymnaea stagnalis neurons, osmolarity increases (upshocks) rapidly elicited large membranous dilations that could be dislodged and pushed around inside the cell with a microprobe. Subsequent osmolarity decreases (downshocks) caused these vacuole-like dilations (VLDs) to disappear. Additional upshock/downshock perturbations resulted in repeated appearance/disappearance (formation/reversal) of VLDs at discrete sites. Confocal microscopy indicated that VLDs formed as invaginations of the substrate-adherent surface of the neuron: extracellular rhodamine-dextran entered VLDs as they formed and was expelled during reversal. Our standard VLD-inducing perturbation was: 2-4 min downshock to distilled water, upshock to normal saline. However, a wide range of other osmotic perturbations (involving osmolarities up to 3.5 x normal, perturbations with or without Ca2+, replacement of ions by sucrose) were also used. We concluded that mechanical, not chemical, aspects of the osmo-mechanical shocks drove the VLD formation and reversal dynamics and that extracellular Ca2+ was not required. Following a standard perturbation, VLDs grew from invisible to their full diameter (> 10 microns) in just over a minute. Over the next 0.5-3 hr in normal saline, neurons recovered. Recovery eliminated any visible VLDs and was accompanied by cytoplasmic turmoil around the VLDs. Recovery was prevented by cytochalasin B, brefeldin A and N-ethylmaleimide but not by nocodazole. In striking contrast, these drugs did not prevent repeated VLD formation and reversal in response to standard osmo-mechanical perturbations; VLD disappearance during reversal and during recovery are different. The osmo-mechanical changes that elicited VLDs may, in an exaggerated fashion, mimic tension changes in extending and retracting neurites. In this context we postulate: (a) the trafficking or disposition of membrane between internal stores and plasma membrane is mechanosensitive, (b) normally, this mechanosensitivity provides an "on demand" system by which neurons can accommodate stretch/release perturbations and control cell shape but, (c) given sudden extreme mechanical stimuli, it yields VLDs.

Animals↗

Sensorin-A immunocytochemistry reveals putative mechanosensory neurons in Lymnaea CNS.

The pond snail Lymnaea stagnalis is a useful model system for studying the neural basis of behaviour but the mechanosensory inputs that impact on behaviours such as respiration, locomotion, reproduction and feeding are not known. In Aplysia, the peptide sensorin-A appears to be specific to a class of central mechanosensory neurons. We show that in the Lymnaea central nervous system sensorin-A immunocytochemistry reveals a discrete pattern of staining involving well over 100 neurons. Identifiable sensorin positive clusters of neurons are located in the buccal and cerebral ganglia, and a single large neuron is immunopositive in each pedal ganglion. These putative mechanosensory neurons are not in the same locations as previously identified motoneurons, interneurons or neurosecretory cells. As would be expected for a mechanoafferent, sensorin positive fibres were found in nerve tracts innervating the body wall. This study lays the foundation for future electrophysiological and behavioural analysis of these putative mechanosensory neurons.

Animals↗

Pharmacology of stretch-activated K channels in Lymnaea neurones.

1. Single-channel recording was used to describe the pharmacology of stretch-activated K channels in Lymnaea neurones using channel blockers amiloride, tetraethylammonium (TEA), quinidine, gadolinium (Gd) and diltiazem. 2. Amiloride, TEA and quinidine applied to the outside face of the membrane all produced a fast flickery block of stretch-activated K channels. All of these agents were without effect when applied at the inside face at concentrations as high as 10, 200 and 10 mM respectively. Neither Gd nor diltiazem had any effect on stretch-activated K channels extracellularly (100 microM). 3. Amiloride, TEA and quinidine block were voltage-independent with IC50 values at positive (and negative membrane potentials of 2.3 (and 2.0) mM, 48 (and 54) mM and 0.8 (and 0.7) mM respectively. Woodhull plots for TEA and quinidine block confirmed the voltage independence of stretch-activated K channel block by these agents. 4. Hill plots of the amilorde, TEA and quinidine block yield Hill coefficients at positive (and negative) membrane potentials of 1.7 (and 1.5), 1.4 (and 1.2) and 1.5 (and 1.6 mM) respectively. 5. Ethanol (3%) had no apparent effect on stretch-activated K channel kinetics or conductance yet reduced the efficacy of quinidine block. 6. The above pharmacological fingerprint of the stretch-activated K channel is discussed with reference to other K-selective and stretch-activated channels.

Amiloride↗

Population Sizes, Immigration, and Growth of Epiphytic Bacteria on Leaves of Different Ages and Positions of Field-Grown Endive (Cichorium endivia var. latifolia).

Total, fluorescent, and pectolytic epiphytic bacterial population sizes were quantified on leaves of different age groups of broad-leaved endive during field cultivation from leaf emergence until harvest. Greater bacterial population densities (log(inf10) CFU per square centimeter) were observed on outer leaves than on inner leaves of the plants throughout the growing season. These differences were statistically significant for total bacterial populations at all sampling times and were often significant for fluorescent and pectolytic bacterial populations. At harvest, a linear gradient of decreasing densities of epiphytic bacteria from outer (older) to inner (younger) leaves of the head was significant. Leaf age influenced the frequency distribution and variability of bacterial population sizes associated with leaves of broad-leaved endive. Total bacterial population sizes were greater at leaf emergence for leaves emerging during the second half of the cultivation period than for leaves emerging earlier. The size of fluorescent and pectolytic bacterial populations on newly emerged leaves increased throughout the season as plants aged. To assess the importance of plant age on bacterial immigration at leaf emergence, bacterial densities were quantified on leaves emerging simultaneously on plants of different ages. In two of the three experiments, greater bacterial population sizes were observed on leaves emerging on younger plants. This indicates that factors other than an increase in concentration of airborne bacteria can lead to increases in population sizes at leaf emergence as plants age in the field. Results of leaf pruning experiments suggested that adjacent leaves may act as a barrier for immigration of fluorescent bacteria on newly emerged leaves. Survival of an inoculated strain of Pseudomonas fluorescens on newly emerged leaves generally did not vary with the age of plants. However, these effects were not consistent among experiments, suggesting that interactions among micro- and macroenvironmental conditions, physiological condition of leaves, and accessibility of leaves to airborne bacteria are important in controlling epiphytic bacterial population sizes.

Journal Article↗

Recovery of function, peripheral sensitization and sensory neurone activation by novel pathways following axonal injury in Aplysia californica.

Recovery of behavioural and sensory function was examined following unilateral pedal nerve crush in Aplysia californica. Nerve crush that transected all axons connecting the tail to the central nervous system (CNS) eliminated the ipsilateral tail-evoked siphon reflex, whose sensory input travels in the crushed tail nerve (p9). The first reliable signs of recovery of this reflex were observed within 1 week, and most animals displayed tail-evoked siphon responses within 2 weeks. Wide-dynamic-range mechanosensory neurons with somata in the ventrocaudal (VC) cluster of the ipsilateral pleural ganglion exhibited a few receptive fields (RFs) on the tail 3 weeks after unilateral pedal nerve crush, indicating that the RFs had either regenerated or been reconnected to the central somata. These RFs were smaller and sensitized compared with corresponding RFs on the contralateral, uncrushed side. Centrally conducted axon responses of VC sensory neurones to electrical stimulation distal to the nerve crush site did not reappear until at least 10 days after the crush. Because the crush site was much closer to the CNS than to the tail, the failure of axon responses to be restored earlier than the behavioural responses indicates that early stages of reflex recovery are not due to regeneration of VC sensory neurone axons into the tail. Following nerve crush, VC sensory neurones often could be activated by stimulating central connectives or peripheral nerves that do not normally contain the sensory neurone's axons. These results suggest that recovery of behavioral function after nerve injury involves complex mechanisms, including regenerative growth of axotomized VC sensory neurones, sensitization of regenerating RFs and sprouting of VC sensory neurone fibres within the CNS. Furthermore, the rapidity of behavioural recovery indicates that its initial phases are mediated by additional mechanisms, perhaps centripetal regeneration of unidentified sensory neurones having peripheral somata, or transient reconnection of proximal and distal stumps of axotomized VC cells.

Animals↗

Peripheral regeneration and central sprouting of sensory neurone axons in Aplysia californica following nerve injury.

Morphological methods were used to examine injury-induced growth of peripheral and central axons of nociceptive mechanosensory neurones in the ventrocaudal (VC) clusters of the pleural ganglia of Aplysia californica. Pedal nerve crush transected all axons in the nerve while leaving the overlying sheath largely intact. Immunohistochemical staining was performed with an antibody to a sensory-neurone-specific peptide, sensorin-A. Following bilateral crush of pedal nerve p9, which innervates the tail, sensorin-A immunofluorescence was lost distal to the crush site within 2 days. Fine immunopositive fibres began to invade the crush region within 5 days. These fibres arborized in the crush region and gradually extended down the crushed nerve. Immunopositive fibres were found near the tail within 3 weeks. Similar results were obtained after injecting individual sensory neurone somata in the tail/p9 region of the VC cluster with biocytin. Biocytin injections and horseradish peroxidase injections 3 weeks after ipsilateral pedal nerve crush revealed new fibres projecting rostrally from the tail/p9 region of the VC cluster and entering the pleural-cerebral and pleural-abdominal connectives. Such projections were never observed in control, uncrushed preparations. These results demonstrate that nerve injury triggers extensive growth of both peripheral and central processes of the VC sensory neurones.

Animals↗

A putative nicotine pump at the metabolic blood-brain barrier of the tobacco hornworm.

In mammals, P-glycoprotein immunostaining at the blood-brain barrier has implicated the multidrug pump in the restricted movement of many cytotoxic agents into the central nervous system (CNS). Since many insects require a sophisticated blood-brain barrier system to protect their CNS from plant-derived neurotoxins, we have investigated the possibility that a P-glycoprotein homolog constitutes a component of the insect blood-brain barrier. We have used the nicotine-resistant tobacco hornworm (Manduca sexta) to address this issue. Manduca has been previously shown, in physiological studies, to have an alkaloid (nicotine/morphine/atropine) pump at its excretory malpighian tubules. We show (1) that the tubules are P-glycoprotein immunopositive, (2) that Manduca has a metabolic blood-brain barrier for nicotine, (3) that the barrier co-localizes with P-glycoprotein immunostaining, and (4) that detoxifying enzymes as well as the nicotine pump are likely to account for the metabolic blood-brain barrier to nicotine. These findings may provide insights on two major fronts, the troublesome problem of multi-insecticide resistance, a phenomenon that parallels multidrug resistance in tumor cells, and the problem of tolerance to addictive neuroactive drugs like nicotine or morphine.

ATP Binding Cassette Transporter, Subfamily B, Mem↗