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Biomedical subjects

G R Stewart

Publications and source records attributed to G R Stewart.

At least 55 records · Page 3Linked to original sources

Effect of Potassium Levels on the Stomatal Behavior of the Hemi-Parasite Striga hermonthica.

The hemi-parasite Striga hermonthica, exhibits an anomalous pattern of stomatal response, stomata remaining open in darkness and when subjected to water stress. This suggests irregularity in stomatal response due to malfunction of the stomatal mechanism. To test this suggestion guard cells were isolated from the effects of surrounding cells, by incubating epidermal strips at low pH. These stomata responded rapidly to low CO(2) concentrations, darkness, and ABA. Thus, a paradox exists between stomatal behavior observed in whole leaves and that in isolated guard cells. However, when incubated in the presence of high potassium concentrations (>200 millimolar KCl) stomatal responses in epidermal strips resembled those found in whole leaves, with enhanced opening and reduced closing responses. It is suggested that the anomalous behavior of stomata in Striga and other leafy hemiparasites can be explained by the modulatory effects of high potassium concentrations which accumulate in the leaves as a consequence of high transpiration rates and the lack of a retranslocation system.

Journal Article↗

Comparison of behavioral effects of nucleus basalis magnocellularis lesions and somatosensory cortex ablation in the rat.

Cholinergic neurons in the nucleus basalis region of the forebrain project to various portions of the cerebral cortex, including somatosensory cortex. Degeneration of these neurons and their cortical projections is a major feature of the neuropathology of Alzheimer's disease. Injecting an excitotoxin into the basal forebrain to destroy nucleus basalis neurons provides a potentially useful animal model for studying the role of these neurons in Alzheimer's disease. Previously, we demonstrated that rats with nucleus basalis excitotoxin lesions performed poorly on a tactile discrimination task and on a test of working memory. In an effort to clarify further the role of impaired memory versus other types of impairment (e.g. disrupted somatosensory processing due to cholinergic deafferentation of somatosensory cortex), we compared a group of rats with bilateral nucleus basalis excitotoxin lesions and a group with bilateral somatosensory cortical ablations on a variety of behavioral tasks. Rats with nucleus basalis lesions performed as well as controls on a battery of neurological tests but exhibited increased emotionality unlike rats with somatosensory cortical ablations which performed poorly on the battery but were not hyperemotional. The two lesion groups were impaired significantly and to a comparable degree in performing two-choice tactile discriminations in a T-maze. In contrast, only rats with nucleus basalis lesions showed deficits in working memory as tested in an eight-arm radial maze. Both lesion groups performed comparably to sham controls on a test of reference memory involving a black/white discrimination in a T-maze. The findings suggest that rats with nucleus basalis lesions manifest disturbances in several of the same spheres (emotionality, somatosensory information processing, memory) that are disrupted in Alzheimer's disease and further confirm the utility of the excitotoxin lesion approach for studying the pathophysiology of Alzheimer's disease.

Animals↗

Prophylactic cranial irradiation for limited non-small cell lung cancer.

Seventy-three patients with biopsy-proven limited non-small cell lung cancer (NSCLC) were entered on a combined modality study at the University of Washington. Seventy-five percent (55 of 73) of the patients had a histologic diagnosis of adenocarcinoma or large cell carcinoma, whereas 25% (18 of 73) had squamous cell carcinoma. After two cycles of chemotherapy, patients without evidence of progressive disease received prophylactic cranial irradiation (PCI) and chest radiotherapy, followed by two additional cycles of chemotherapy. Brain computed tomography (CT) scans were performed at 3-month intervals after completion of therapy in all patients, and were additionally performed whenever signs or symptoms developed suggesting neurologic dysfunction or recurrent brain disease. Sixty-five patients were treated with PCI. No clinical or CT evidence of recurrence in the brain has developed in patients who completed PCI. PCI appears to be effective in greatly reducing the incidence of brain relapse in patients with limited NSCLC.

Brain↗

Fast neutron radiotherapy--an overview.

Fast neutron radiotherapy has been employed worldwide in the last two decades in the treatment of a variety of advanced malignancies. The basic principles behind neutron treatment will be summarized in this article, along with results to date of a number of clinical trials evaluating the effectiveness and toxicity of neutron radiotherapy.

Clinical Trials as Topic↗

Choroid plexus carcinoma of the fourth ventricle. Report of a case in an infant.

An 8-month-old female infant with choroid plexus carcinoma presenting in the fourth ventricle is described. The patient was initially treated with combination chemotherapy due to her young age, but developed local tumor recurrence 13 months after beginning treatment. Staging studies to detect tumor seeding of the cerebrospinal fluid were negative, and the patient received posterior fossa radiotherapy. Although the primary tumor regressed with radiation treatment, widespread disease throughout the spinal subarachnoid space subsequently developed. Choroid plexus carcinoma has a significant tendency to metastasize through the cerebrospinal fluid, and treatment of the entire neuraxis should be strongly considered for patients with this disease.

Antibodies, Monoclonal↗

Gas exchange characteristics of the sorghum-striga host-parasite association.

Gas exchange characteristics are reported for both members of the sorghum-Striga host-parasite association. Both Striga hermonthica (Del.) Benth and Striga asiatica (L.) Kuntze had transpiration rates considerably in excess of those of sorghum (Sorghum bicolor (L.) Moench, cv CSH1). Stomatal conductance in both Striga spp. showed little response to periods of darkness and moderate water stress. Low rates of net CO(2) fixation and high rates of dark respiration led to no net daily (24 hours) C gain, and Striga would appear to be reliant on its host for photosynthate. Infection of sorghum plants with either S. hermonthica or S. asiatica reduced host photosynthetic capacity. Infected sorghum plants were also more prone to water stress, but reduced rates of CO(2) fixation could not be accounted for in terms of lower stomatal conductance. Lower stomatal conductances were associated with an increase in water use efficiency (WUE) in uninfected sorghum; however, Striga-infected sorghum plants had lower WUE than those of uninfected plants. We suggest that Striga exerts a specific effect on processes affecting C acquisition in sorghum leaves. The water relations of S. hermonthica and S. asiatica are not characteristic of plants growing in semiarid environments and are more likely to reflect the nature of the parasitic life-style. Despite transfer of water and solutes from host to parasite, the reduction in C fixation observed in infected sorghum plants appears to be the major determinant of growth reductions observed in sorghum supporting Striga.

Journal Article↗

Carbon isotope ratios demonstrate carbon flux from c(4) host to c(3) parasite.

Carbon isotope ratios of mature leaves from the C(3) angiosperm root hemiparasites Striga hermonthica (Del.) Benth (-26.7 per thousand) and S. asiatica (L.) Kuntze (-25.6 per thousand) were more negative than their C(4) host, sorghum (Sorghum bicolor [L.] Moench cv CSH1), (-13.5 per thousand). However, in young photosynthetically incompetent plants of S. hermonthica this difference was reduced to less than 1 per thousand. Differences between the carbon isotope ratios of two C(3)-C(3) associations, S. gesnerioides (Willd.) Vatke-Vigna unguiculata (L.) Walp. and Oryza sativa L.-Rhamphicarpa fistulosa (Hochst.) Benth differed by less than 1 per thousand. Theoretical carbon isotope ratios for mature leaves of S. hermonthica and S. asiatica, calculated from foliar gas exchange measurements, were -31.8 and -32.0 per thousand, respectively. This difference between the measured and theoretical delta(13)C-values of 5 to 6 per thousand suggests that even in mature, photosynthetically active plants, there is substantial input of carbon from the C(4) host. We estimate this to be approximately 28% of the total carbon in S. hermonthica and 35% in S. asiatica. This level of carbon transfer contributes to the host's growth reductions observed in Striga-infected sorghum.

Journal Article↗

Fibronectin-like immunoreactivity in the developing cerebral cortex.

In the developing cerebral cortex of the mouse, binding of antibodies directed against the extracellular matrix glycoprotein fibronectin occurs with a distinct temporal and spatial pattern. On the 10th embryonic day (E10), when the wall of the telencephalic vesicle is made up of only the proliferating cells of the ventricular zone, antifibronectin (aFN) binding is restricted to the blood vessels and pia-arachnoid. Fibronectin-like immunoreactivity first appears in the neuropil as small points of immunofluorescence among the earliest postmitotic neurons that form the preplate (E11-12). A short time later (E12-13), aFN immunoreactivity becomes more diffuse but continues to be restricted to the preplate. As newly arriving neurons form the cortical plate within the preplate (E13-14), aFN binding is present in the marginal zone above the cortical plate and in the subplate below it. Both the marginal zone and the subplate contain early afferents and the cells that were previously part of the preplate. Binding of aFN is transient; by E18-19 it has diminished to the point where it is no longer detectable except in the blood vessels and pia-arachnoid. The transient appearance of fibronectin-like immunostaining in the zones that contain early cortical afferents suggests that fibronectin plays a role in forming the migratory pathway for the growth cones of these axons. In this role it may be acting in concert with other extracellular matrix components such as hyaluronectin, glycosaminoglycans, and laminin, which have been shown to have similar spatial distributions.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

N-methylaspartate: an effective tool for lesioning basal forebrain cholinergic neurons of the rat.

The ability of the excitotoxin, N-methyl-D,L-aspartic acid (NMA), to destroy basal forebrain cholinergic (BFC) neurons was evaluated. NMA (100 nmol) was directly injected into the peripallidum, a region containing a proportionately large number of cortically-projecting BFC neurons. Cholineacetyltransferase (ChAT) activity 10 days later was markedly and significantly reduced (up to 62%) in the cortex ipsilateral to the lesion. NMA induced a focal lesion affecting BFC neurons without damaging axons of passage or causing lesions distant from the site of injection. ChAT immunohistochemistry (IHC) was used to directly demonstrate loss of ChAT-positive neurons from the lesion site. This loss persisted at all survival times examined, from 2 days to 7.5 months post-injection.

Animals↗

Cholinergic denervation-induced increase of chelatable zinc in mossy-fiber region of the hippocampal formation.

Chelatable zinc has repeatedly been associated with hippocampal mossy fibers, but the neurobiological functions of the zinc have remained unclear. Zinc is a constituent of the 7S Nerve Growth Factor (NGF) molecule. The hippocampal area is rich in neurotrophic factors and cholinergic deafferentation of the hippocampus appears to activate these neurotrophic factors. In this paper we report that cholinergic deafferentation of the hippocampus causes a transient rise in mossy-fiber zinc, as measured by zinc-dithizonate histochemistry. The changes are not significant until 10 days after the septal lesion, and the zinc returns to control levels by 21-28 days. Dithizonate has an association constant of 10(-15) with zinc, while NGF has an association constant of 10(-11). Thus densitometric measurement of zinc dithizone within the hippocampus may be a quantitative histochemical marker of a zinc-associated nerve growth-like substance. The data support recent findings that nerve growth-like factors increase in the hippocampal area following deafferentation, and in addition suggest that zinc is a critical factor in the regulation of trophic phenomena in the hippocampal formation.

Animals↗

Cytosolic glutamine synthetase in higher plants. A comparative immunological study.

Cytosolic glutamine synthetase (GS1) was purified to homogeneity from etiolated barley leaves by DEAE-Sephacel and hydroxyapatite chromatography, gel filtration and polyacrylamide gel electrophoresis. Specific antibodies against the purified protein were raised by the immunization of rabbits. Immunoprecipitation experiments demonstrated that cytosolic glutamine synthetases isolated from the leaves of different plant species were very similar proteins. Good recognition of other cytosolic glutamine synthetases from roots, root nodular tissue and seeds by barley GS1 antibodies was obtained, suggesting that they too are all quite similar proteins. In contrast, chloroplast glutamine synthetase (GS2) was considered to be a different protein in view of its low level of recognition by barley GS1 antibodies.

Antibody Specificity↗

Occurrence of Isóenzymes of Glutamine Synthetase in the Alga Chlorella kessleri.

Two forms (GS(1) and GS(2)) of glutamine synthetase have been isolated, separated by ion exchange chromatography, and partly characterized from cells of the green alga Chlorella kessleri. Both forms are present in cells grown autotrophically or heterotrophically on various nitrogen sources, but under all nutritional conditions GS(1) was found to be the major isoenzyme present (60-80%). The activity of both isoenzymes was greatest in cells grown under nitrogen-limiting conditions. Both isoenzymes have molecular weights in the range 340 to 350,000 daltons. GS(1) was found to have a greater thermostability than GS(2): GS(1) was stable at 30 degrees C while GS(2) lost 95% of its activity in 30 minutes. GS(1) was much less sensitive to thiol reactive reagents than GS(2).

Journal Article↗

Proline and glycine betaine influence protein solvation.

Glutamine synthetase from barley (Hordeum distichum L.) is precipitated by polyethylene glycol (PEG). Proline, in a concentration-dependent manner, reduces the amount of enzyme precipitated by PEG, although the effect of the imino acid can be counteracted by raising the level of PEG. The effect of PEG is a function of mer number and concentration and the influence of both elements can be ameliorated by proline. PEG-induced enzyme precipitation is a function of pH, as is its interaction with both proline and betaine in the reaction. The lack of effect of amount of enzyme on the proline and PEG effects supports the conclusion that, in this system, proline and PEG do not function through interaction with the protein. Other compounds, such as glycine, glucose, and sucrose, can decrease the PEG-induced precipitation of the enzyme, although glycerol was not active under the conditions employed.The results are consistent with the proposition that a protein-containing system in which high concentrations of proline and/or betaine are present, is better ;protected' against the biologically unfavorable consequences of dehydration-induced thermodynamic perturbation.

Journal Article↗

Nitrogen Assimilation in Mycorrhizas : Ammonium Assimilation in the N-Starved Ectomycorrhizal Fungus Cenococcum graniforme.

Ammonium assimilation was followed in N-starved mycelia from the ectomycorrhizal Ascomycete Cenococcum graniforme. The evaluation of free amino acid pool levels after the addition of 5 millimolar NH(4) (+) indicated that the absorbed ammonium was assimilated rapidly. Post-feeding nitrogen content of amino acids was very different from the initial values. After 8 hours of NH(4) (+) feeding, glutamine accounted for the largest percentage of free amino acid nitrogen (43%). The addition of 5 millimolar methionine sulfoximine (MSX) to NH(4) (+)-fed mycelia caused an inhibition of glutamine accumulation with a corresponding increase in glutamate and alanine levels.Using (15)N as a tracer, it was found that the greatest initial labeling was into glutamine and glutamate followed by aspartate, alanine, and ornithine. On inhibiting glutamine synthetase using MSX, (15)N enrichment of glutamate, alanine, aspartate, and ornithine continued although labeling of glutamine was quite low. Moreover, the incorporation of (15)N label in insoluble nitrogenous compounds was lower in the presence of MSX. From the composition of free amino acid pools, the (15)N labeling pattern and effects of MSX, NH(4) (+) assimilation in C. graniforme mycelia appears to proceed via glutamate dehydrogenase pathway. This study also demonstrates that glutamine synthesis is an important reaction of ammonia utilization.

Journal Article↗

Ammonia Assimilation in the Roots of Nitrate- and Ammonia-Grown Hordeum Vulgare (cv Golden Promise).

(15)N kinetic labeling studies were performed on seedlings of Hordeum vulgare L. var. Golden Promise growing under steady state conditions. Patterns of label incorporation in the pools of nitrogen compounds of roots fed [(15)N]ammonium were compared with computer-simulated labeling curves. The data were found to be quantitatively consistent with a three-compartment model in which ammonium is assimilated solely into the amide-N of glutamine. Labeling data from roots fed [(15)N]nitrate were also found to be at least qualitatively consistent with the assimilation of ammonia into glutamine. Methionine sulfoximine almost completely blocked the incorporation of (15)N label into the amino acid pools of barley roots fed [(15)N]nitrate. These observations suggest that ammonia assimilation occurs solely via the glutamine synthetase/glutamate synthase pathway in both nitrate- and ammonia-grown barley roots.

Journal Article↗

Action of Inhibitors of Ammonia Assimilation on Amino Acid Metabolism in Hordeum vulgare L. (cv Golden Promise).

Barley (Hordeum vulgare L. cv Golden Promise) plants were grown in a continuous culture system in which the root and shoot ammonia and amino acid levels were constant over a 6-hour experimental period. Methionine sulfoximine (MSO), 1 millimolarity when added to the culture medium, caused a total inactivation of root glutamine synthetase with little effect on the shoot enzyme. Root ammonia levels increased and glutamine levels decreased, irrespective of whether the plants were grown in 1 millimolar nitrate or 1 millimolar ammonia. Levels of glutamate, aspartate, serine, threonine, and asparagine all increased. There was little alteration in the amino acid and ammonia levels in the shoot, suggesting that MSO is not rapidly transported.The addition of azaserine (25 micrograms per milliliter) to nitrate-grown plants caused a rapid increase in root ammonia, glutamine, and serine levels with a corresponding decrease in glutamate, aspartate, and alanine. Glutamine levels also increased in the shoot.The in vivo effect of MSO and azaserine was as would be predicted by their known in vitro inhibitory action if the glutamine synthetase/glutamate synthase pathway of ammonia assimilation was in operation.

Journal Article↗

Glutamine Synthetases of Higher Plants : Evidence for a Specific Isoform Content Related to Their Possible Physiological Role and Their Compartmentation within the Leaf.

The chromatographic properties of glutamine synthetase isoforms have been investigated in a wide range of higher plant leaves and shoots using ion exchange chromatography. Different patterns of glutamine synthetase isoform content were observed. Among higher plants, four patterns or groups could be recognized. The first group is characterized by having only cytosolic glutamine synthetase, whereas the second group is distinguished by having only chloroplastic glutamine synthetase. The third group is characterized by cytosolic glutamine synthetase being a minor component of the total leaf glutamine synthetase activity. The fourth group is distinct from the other groups in having high cytosolic and chloroplast glutamine synthetase activity. Immunological studies have been undertaken on a few species from each group to identify unambiguously both cytosolic and chloroplastic glutamine synthetases.

Journal Article↗