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Biomedical subjects

T J Andrews

Publications and source records attributed to T J Andrews.

At least 37 records · Page 2Linked to original sources

The perception of transparent three-dimensional objects.

When the proximal and distal elements of wire-frame cubes are conflated, observers perceive illusory structures that no longer behave veridically. These phenomena suggest that what we normally see depends on visual associations generated by experience. The necessity of such learning may explain why the mammalian visual system is subject to a prolonged period of plasticity in early life, when novel circuits are made in enormous numbers.

Aging↗

Correlated size variations in human visual cortex, lateral geniculate nucleus, and optic tract.

We have examined several components of the human visual system to determine how the dimensions of the optic tract, lateral geniculate nucleus (LGN), and primary visual cortex (V1) vary within the same brain. Measurements were made of the cross-sectional area of the optic tract, the volumes of the magnocellular and parvocellular layers of the LGN, and the surface area and volume of V1 in one or both cerebral hemispheres of 15 neurologically normal human brains obtained at autopsy. Consistent with previous observations, there was a two- to threefold variation in the size of each of these visual components among the individuals studied. Importantly, this variation was coordinated within the visual system of any one individual. That is, a relatively large V1 was associated with a commensurately large LGN and optic tract, whereas a relatively small V1 was associated with a commensurately smaller LGN and optic tract. This relationship among the components of the human visual system indicates that the development of its different parts is interdependent. Such coordinated variation should generate substantial differences in visual ability among humans.

Adult↗

Side reactions catalyzed by ribulose-bisphosphate carboxylase in the presence and absence of small subunits.

The large subunit core of ribulose-bisphosphate carboxylase from Synechococcus PCC 6301 expressed in Escherichia coli in the absence of its small subunits retains a trace of carboxylase activity (about 1% of the kcat of the holoenzyme) (Andrews, T. J (1988) J. Biol. Chem. 263, 12213-12219). During steady-state catalysis at substrate saturation, this residual activity diverted approximately 10% of the reaction flux to 1-deoxy-D-glycero-2,3-pentodiulose-5-phosphate as a result of beta elimination of inorganic phosphate from the first reaction intermediate, the 2,3-enediol form of ribulose bisphosphate. This indicates that the active site's ability to stabilize and/or retain this intermediate is compromised by the absence of small subunits. Epimerization and isomerization of the substrate resulting from misprotonation of the enediol intermediate were not significantly exacerbated by lack of small subunits. The residual carboxylating activity partitioned product between pyruvate and 3-phosphoglycerate in a ratio similar to that of the holoenzyme, indicating that stablization of the penultimate three-carbon aci-acid intermediate is not perturbed by lack of small subunits. The underlying instability of the five-carbon enediol intermediate was revealed, even with the holoenzyme, under conditions designed to lead to exhaustion of substrate CO2 (and O2). When carboxylation (and oxygenation) stalled upon exhaustion of gaseous substrate, both spinach and Synechococcus holoenzymes continued slowly to beta eliminate inorganic phosphate from and to misprotonate the enediol intermediate. With carboxylation and oxygenation blocked, the products of these side reactions of the enediol intermediate accumulated to readily detectable levels, illustrating the difficulties attendant upon ribulose-P2 carboxylase's use of this reactive species as a catalytic intermediate.

Borates↗

Can illusory motion disrupt tracking real motion?

When rotating stripes or other periodic stimuli cross the retina at a critical rate, a reversal in the direction of motion of the stimuli is often seen. This illusion of motion perception was used to explore the roles of retinal and perceived motion in the generation of optokinetic nystagmus. Here we show that optokinetic nystagmus is disrupted during the perception of this illusion. Thus, when perceived and actual motion are in conflict, subjects fail to track the veridical movement. This observation suggests that the perception of motion can directly influence optokinetic nystagmus, even in the presence of a moving retinal image. A conflict in the neural representation of motion in different brain areas may explain these findings.

Humans↗

Ribulose-1,5-bisphosphate carboxylase/oxygenase activase deficiency delays senescence of ribulose-1,5-bisphosphate carboxylase/oxygenase but progressively impairs its catalysis during tobacco leaf development.

Transgenic tobacco (Nicotiana tabacum L. cv W38) plants with an antisense gene directed against the mRNA of ribulose-1,5-biphosphate carboxylase/oxygenase (Rubisco) activase grew more slowly than wild-type plants in a CO2-enriched atmosphere, but eventually attained the same height and number of leaves. Compared with the wild type, the anti-activase plants had reduced CO2 assimilation rates, normal contents of chlorophyll and soluble leaf protein, and much higher Rubisco contents, particularly in older leaves. Activase deficiency greatly delayed the usual developmental decline in Rubisco content seen in wild-type leaves. This effect was much less obvious in another transgenic tobacco with an antisense gene directed against chloroplast-located glyceraldehyde-3-phosphate dehydrogenase, which also had reduced photosynthetic rates and delayed development. Although Rubisco carbamylation was reduced in the anti-activase plants, the reduction was not sufficient to explain the reduced photosynthetic rate of older anti-activase leaves. Instead, up to a 10-fold reduction in the catalytic turnover rate of carbamylated Rubisco in vivo appeared to be the main cause. Slower catalytic turnover by carbamylated Rubisco was particularly obvious in high-CO2-grown leaves but was also detectable in air-grown leaves. Rubisco activity measured immediately after rapid extraction of anti-activase leaves was not much less than that predicted from its degree of carbamylation, ruling out slow release of an inhibitor from carbamylated sites as a major cause of the phenomenon. Nor could substrate scarcity or product inhibition account for the impairment. We conclude that activase must have a role in vivo, direct or indirect, in promoting the activity of carbamylated Rubisco in addition to its role in promoting carbamylation.

Carbon Dioxide↗

Autonomic nervous system as a model of neuronal aging: the role of target tissues and neurotrophic factors.

The aim of this study was to determine the role of target tissues and neurotrophic factors in the growth and atrophy of autonomic neurons during development and aging. Using quantitative neuroanatomical techniques, it is shown that, although axonal and dendritic growth is apparent throughout postnatal development, different patterns of growth are found in autonomic neurons innervating different target tissues. For example, sympathetic neurons innervating the submandibular gland continue to grow well into maturity, but those innervating the iris cease net growth early in postnatal development. Similarly, although neuronal atrophy was observed in aged autonomic ganglia, this was not a general phenomenon but was specific to neurons innervating particular target tissues. Sympathetic neurons innervating the middle cerebral artery showed significant axonal and dendritic atrophy in old age, whereas neurons innervating the iris were morphologically unchanged. The trophic influence of peripheral target tissues on their innervating neurons has been shown to decline in old age possibly as a result of decreased availability of target-derived neurotrophic factors such as nerve growth factor (NGF) [Gavazzi et al. (1992) Neuroreport, 3:717-720]. Therefore, in an attempt to reverse neuronal atrophy where it occurred, NGF was infused via miniosmotic pumps over the peripheral axons of aged neurons. NGF induced increases in soma size, dendritic length and axonal arborization. However, in contrast to young adult neurons, no increase in the number of dendritic branch points or primary dendrites was observed, suggesting that some aspects of neuronal plasticity are impaired in old age. In sum, these results show a range of age- and target-specific differences in the axonal and dendritic morphology of autonomic neurons that may result from differing trophic interactions with their target tissues.

Aging↗

Target-specific differences in the dendritic morphology and neuropeptide content of neurons in the rat SCG during development and aging.

Our purpose in this work was to investigate the role of target tissues in the regulation of dendritic morphology from sympathetic neurons during development and aging. Neurons were retrogradely labeled from three targets, the iris, the submandibular gland (SMG), and the middle cerebral artery (MCA). They were then fixed and intracellularly injected to demonstrate their dendritic arborizations. Dendritic geometry varied quantitatively in sympathetic neurons innervating different target tissues at all stages of development. Neurons innervating the iris had the largest cell bodies and most extensive dendritic arborizations, whereas the vasomotor neurons were the smallest. The number of primary dendrites, however, did not vary significantly between the different neuronal populations. The growth of dendritic arborizations during postnatal development and their atrophy in old age were not concordant in the different neuron populations we studied. Neurons innervating the MCA and the iris ceased dendritic growth early in postnatal development, whereas the dendritic complexity of neurons supplying the SMG increased well into adulthood. By contrast, dendritic atrophy was seen in aged MCA- and SMG-projecting neurons but not in those innervating the iris, suggesting, with other evidence, correlated and distinct patterns of growth and atrophy in axons and dendrites of mature sympathetic neurons projecting to different targets. Swollen dendrites and protuberances on cell soma were a prominent feature of aged neurons. In addition to the target-specific variation in neuronal morphology, we observed diversity in neurotransmitter phenotype. For example, neuropeptide Y was expressed in iridial but not SMG-projecting neurons. These results show a range of age- and target-specific differences in the dendritic morphology and neuropeptide content of sympathetic neurons that may be a result of differing trophic interactions with their target tissues.

Aging↗

Temporal events in cyclopean vision.

The majority of neurons in the primary visual cortex of primates can be activated by stimulation of either eye; moreover, the monocular receptive fields of such neurons are located in about the same region of visual space. These well-known facts imply that binocular convergence in visual cortex can explain our cyclopean view of the world. To test the adequacy of this assumption, we examined how human subjects integrate binocular events in time. Light flashes presented synchronously to both eyes were compared to flashes presented alternately (asynchronously) to one eye and then the other. Subjects perceived very-low-frequency (2 Hz) asynchronous trains as equivalent to synchronous trains flashed at twice the frequency (the prediction based on binocular convergence). However, at higher frequencies of presentation (4-32 Hz), subjects perceived asynchronous and synchronous trains to be increasingly similar. Indeed, at the flicker-fusion frequency (approximately 50 Hz), the apparent difference between the two conditions was only 2%. We suggest that the explanation of these anomalous findings is that we parse visual input into sequential episodes.

Animals↗

The wagon wheel illusion in movies and reality.

Wheels turning in the movies or in other forms of stroboscopic presentation often appear to be rotating backward. Remarkably, a similar illusion is also seen in continuous light. The occurrence of this perception in the absence of intermittent illumination suggests that we normally see motion, as in movies, by processing a series of visual episodes.

Eye Movements↗

Objective and subjective factors in the disproportionate referral of children for academic problems.

R. S. Drabman, K. J. Tarnowski, and P. A. Kelly (1987) and K. J. Tarnowski, D. F. Anderson, R. S. Drabman, and P. A. Kelly (1990) examined children's month of birth in relation to referral for psychological services and found that younger children in the classroom were disproportionately referred for services. No differences were found between younger and older students on standardized measures of intelligence or academic achievement. Results of a replication and extension of these studies indicated (a) that younger children in the classroom were referred at a disproportionately higher rate, (b) that the referral pattern could not be explained by differences in children's competencies, (c) that Caucasian students were referred at disproportinately higher rates than minority students, and (d) a trend in which the proportionate referral rate of students as height or weight increased. Results are discussed within the context of teacher expectancies.

Adolescent↗

Manual asymmetry and handedness.

Volumetric measurements show that right-handed individuals have larger right hands than left hands. In contrast, the hands of left-handers are much more nearly symmetrical. Based on what is known about trophic interactions between neurons and targets, these findings predict a corresponding asymmetry of the relevant parts of the sensorimotor system in right-handers. The lack of an opposite-hand asymmetry among left-handers further implies that right- and left-handed phenotypes do not arise according to the same developmental rules.

Adult↗

Mutations of an active site threonyl residue promote beta elimination and other side reactions of the enediol intermediate of the ribulosebisphosphate carboxylase reaction.

The side chain of residue threonine 65 within the active site of ribulosebisphosphate carboxylase participates in a network of hydrogen bonds and ionic interactions involving the phosphate moiety attached to C-1 of the substrate. This residue was replaced with serine, alanine, and valine in the enzyme from Synechococcus PCC 6301. The mutant enzymes were stable, expressed abundantly by Escherichia coli, and retained the ability to form gel-filterable complexes with the reaction-intermediate analog, 2'-carboxyarabinitol-1,5-bisphosphate. The substitutions reduced the kcat/Km(CO2) (where kcat is the substrate-saturated turnover rate) of the enzyme from 17- to 340-fold with the more radical substitutions causing more severe reductions. The CO2/O2 specificity also deteriorated progressively, the valine replacement causing a 2.3-fold reduction. In concert with these changes, a compound tentatively identified as 1-deoxy-D-glycero-2,3-pentodiulose-5-phosphate, the product of beta elimination of the 2,3-enediol(ate) intermediate of the catalytic reaction, appeared among the reaction products in progressively increasing amounts. In the case of the valine substitution, it comprised 13% of the ribulose bisphosphate consumed. The mutant enzymes also partitioned more of their reaction flux to pentulose bisphosphate isomers of ribulose bisphosphate. By contrast, the diversion of carboxylated product to pyruvate, as a result of beta elimination of the three-carbon aci-carbanion intermediate of the carboxylation reaction, was ameliorated by the replacements, the valine mutant showing a 5-fold improvement in this parameter. These observations focus attention on a geometric conflict which exists between the requirements for stabilization of the 5-carbon enediol(ate) and 3-carbon aci-carbanion intermediates. This conflict must be resolved by a change in the angle of the C-1/bridge oxygen bond during each catalytic cycle. The network of hydrogen bonds involving the side chain of threonine 65 must play a crucial role in facilitating reaction of the enediol(ate) with the gaseous substrate and in shepherding this subsequent movement.

Amino Acid Sequence↗

Nerve growth factor enhances the dendritic arborization of sympathetic ganglion cells undergoing atrophy in aged rats.

We have investigated whether dendritic growth can be induced from sympathetic neurons of aged rats by the application of exogenous nerve growth factor to their target tissues. A previous study showed that significant dendritic atrophy (19%) occurs during aging in the sympathetic neurons innervating the middle cerebral artery and that dendritic atrophy correlated with loss of axon collaterals in the same population of neurons. Using retrograde tracing in conjunction with intracellular injection of fixed tissue and confocal microscopy, we now demonstrate that infusion of nerve growth factor over the peripheral processes of the same neurons from aged rats induces significant dendritic (45%) and cell body (60%) growth. However, not all aspects of the dendritic arborization were affected. Primary dendrites and branch points were not altered by nerve growth factor. In contrast, nerve growth factor induced a significant increase in the number of primary dendrites and branch points (100%) of neurons from young adults. Our results demonstrate that sympathetic neurons undergoing dendritic atrophy during aging can exhibit significant dendritic growth in response to the in vivo infusion of nerve growth factor, although the lack of regenerative response displayed by some parts of the dendritic tree leads us to believe that these neurons also show signs of reduced plasticity.

Aging↗

The effect of age on dendrites in the rat superior cervical ganglion.

Intracellular injection of a biotinylated probe in fixed superior cervical ganglia followed by confocal microscopy was used to investigate the effects of age on the dendritic arborisation of sympathetic neurons in rats aged 6 wk (young adult), 7 months (fully grown adult) and 24 months (aged). In accordance with other studies considerable dendritic growth was observed during postnatal development. However, in old age dendritic growth did not continue, and significant atrophy was observed. Quantitation of neuronal morphology showed significant reductions in soma size, total dendritic length, number of branch points and total area of dendritic arborisation in old age. Unexpectedly, significant reductions in the numbers of primary dendrites were observed in maturity and in old age. Concomitant with this atrophy there was an increase in age-related morphological abnormalities. The similarities between the atrophy and dendritic abnormalities shown by our aged neurons and those seen in other studies of young adult sympathetic neurons following axotomy or trophic factor deprivation are discussed.

Aging↗

In vivo infusion of NGF induces the organotypic regrowth of perivascular nerves following their atrophy in aged rats.

The aim of this study was to investigate whether NGF could reverse the nerve fiber atrophy exhibited by some neurons in old age. We used quantitative immunohistochemical techniques to investigate how the autonomic nerves that supply different blood vessels are affected by aging. Age changes in the nerve plexus were not widespread but were specific to particular vascular targets. Furthermore, where nerve fiber atrophy did occur, it affected different populations of nerves; specifically, sympathetic nerve fibers were lost from the middle cerebral artery while nonsympathetic nerves were lost from the tail vein. Peripheral target tissues have been shown to have a trophic influence on the pattern and density of their innervation, which declines in old age (Gavazzi et al., 1992), possibly as a result of decreased availability of neurotrophic factors such as NGF. Consequently, in an attempt to reverse nerve fiber atrophy, we used miniosmotic pumps to infuse NGF for 2 weeks over involuting nerve fibers in the middle cerebral artery of freely moving aged rats. Our results show that exogenous NGF can induce organotypic neurite outgrowth from aged neurons undergoing nerve fiber atrophy. Moreover, NGF can induce a change in the pattern of the old nerve plexus to one similar to that seen in young animals. NGF had different effects on the extent and manner of neurite outgrowth in young and aged nerves. NGF induced greater nerve growth in old compared to young nerves, causing old nerves to grow within existing nerve bundles and to sprout new nerve fibers, in contrast to young nerves, which tended to grow only within existing nerve bundles. In conclusion, this study shows the potential of exogenous NGF to reverse age-related changes in neuronal morphology.

Aging↗

The source and characteristics of chemiluminescence associated with the oxygenase reaction catalyzed by Mn(2+)-ribulosebisphosphate carboxylase.

We confirm the observation of Mogel and McFadden (Mogel, S.N., and McFadden, B. A. (1990) Biochemistry 29, 8333-8337) that ribulosebisphosphate carboxylase/oxygenase (rubisco) exhibits chemiluminescence while catalyzing its oxygenase reaction in the presence of Mn2+. However, our results with the spinach and Rhodospirillum rubrum enzymes differ markedly in the following respects. 1) Chemiluminescence intensity was directly proportional to enzyme concentration and behaved as if representing the rate of oxygenase catalysis. 2) The wavelength spectrum peaked at about 770 nm and extended beyond 810 nm. This seems inconsistent with chemiluminescence generated by simultaneous decay of pairs of singlet O2 molecules. It is consistent with manganese(II) luminescence and we discuss its possible sources. The time course of chemiluminescence (resolution, 0.25 s) was distinctively different for spinach and R. rubrum enzymes during the initial 5 s of catalysis, with the bacterial enzyme exhibiting a pronounced initial "burst." Chemiluminescence by the spinach enzyme responded to substrate concentrations in a manner consistent with known oxygenase properties, exhibiting Michaelis-Menten kinetics with ribulose-1,5-bisphosphate (Km 400 nM). Chemiluminescence required carbamylated enzyme with Mn2+ bound at the active site (activation energy, -57.1 KJ.mol-1). As an indicator of oxygenase activity, chemiluminescence represents an improvement over oxygen electrode measurements in response time and sensitivity by factors of at least 100.

Kinetics↗

Reduction of ribulose biphosphate carboxylase activase levels in tobacco (Nicotiana tabacum) by antisense RNA reduces ribulose biphosphate carboxylase carbamylation and impairs photosynthesis.

The in vivo activity of ribulose-1,5-biphosphate carboxylase/oxygenase (Rubisco) is modulated in response to light intensity by carbamylation of the active site and by the binding of sugar phosphate inhibitors such as 2'-carboxyarabinitol-1-phosphate (CA 1P). These changes are influenced by the regulatory protein Rubisco activase, which facilitates the release of sugar phosphates from Rubisco's catalytic site. Activase levels in Nicotiana tabacum were reduced by transformation with an antisense gene directed against the mRNA for Rubisco activase. Activase-deficient plants were photosynthetically impaired, and their Rubisco carbamylation levels declined upon illumination. Such plants needed high CO2 concentrations to sustain reasonable growth rates, but the level of carbamylation was not increased by high CO2. The antisense plants had, on average, approximately twice as much Rubisco as the control plants. The maximum catalytic turnover rate (k cat) of Rubisco decreases in darkened tobacco leaves because of the binding of CA 1P. The dark-to-light increase in k cat that accompanies CA 1P release occurred to similar extents in antisense and control plants, indicating that normal levels of activase were not essential for CA 1P release from Rubisco in the antisense plants. However, CA 1P was released in the antisense plants at less than one-quarter of the rate that it was released in the control plants, indicating a role for activase in accelerating the release of CA 1P.

Base Sequence↗