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Biomedical subjects

M Stewart

Publications and source records attributed to M Stewart.

At least 181 records · Page 10Linked to original sources
PubMed1997

Nuclear protein import is decreased by engineered mutants of nuclear transport factor 2 (NTF2) that do not bind GDP-Ran.

Nuclear transport factor 2 (NTF2) is associated with the translocation stage of nuclear protein import and binds both to nuclear pore proteins (nucleoporins) containing phenylalanine-rich repeats and to the Ras family GTPase Ran. In this study we probed the role of the NTF2-Ran interaction in nuclear protein import using site-directed mutants of NTF2 that interfere with its interaction with GDP-Ran. The design of these mutants was based on the X-ray crystal structure of NTF2 and was concentrated on conserved residues in and around the molecule's hydrophobic cavity. The mutant NTF2 cDNAs were expressed in Escherichia coli. Purified mutant proteins retained the interaction with FxFG-repeat nucleoporins, but several mutants in the negatively charged residues that surround the NTF2 cavity or in residues in the cavity itself were unable to bind GDP-Ran in vitro. The crystal structure of the E42K mutant protein showed significant structural changes only in this side-chain, indicating that it participated directly in the interaction with GDP-Ran. In permeabilised cell nuclear protein import assays, only wild-type NTF2 and mutants that bound GDP-Ran were functional. Furthermore, when the NTF2 E42K and D92N/D94N NTF2 mutants that failed to bind GDP-Ran in vitro were substituted for the chromosomal yeast NTF2, the yeast cells became non-viable, whereas yeast substituted with wild-type human NTF2 remained viable. We conclude that interaction between NTF2 and GDP-Ran is important for efficient nuclear protein import.

Animals↗
PubMed1997

Antidromic and orthodromic responses by subicular neurons in rat brain slices.

The subiculum forms part of the region of transition between hippocampus and entorhinal cortex and is one of the primary output structures of the hippocampal formation. Intracellular recordings from subicular bursting and non-bursting cell types and field potential recordings were taken in horizontal slices from rat brains. The inputs and outputs of the two cell types were studied for the purpose of reinforcing or refuting the dichotomy proposed on the basis of membrane properties. Some bursting cells were antidromically activated by stimuli applied to the superficial or deep layers of presubiculum, but never by stimuli applied to deep layers of medial entorhinal cortex (dMEC). Some non-bursting subicular neurons were antidromically activated by stimuli applied to dMEC, but never by stimuli applied to presubiculum. Antidromic population events in subiculum were single spikes when deep MEC was stimulated, but were bursts when presubiculum was stimulated, even in the presence of glutamate receptor antagonists. Population bursts consist of 2 or more population spikes with peak to peak intervals of approximately 5 ms. That population bursts occur in slices where excitatory transmission is blocked suggests that such population bursts reflect coincident bursts by individual neurons. Short-latency (< 5 ms) excitatory postsynaptic potentials (EPSPs) were evoked in both subicular cell types in response to single entorhinal, presubicular and CA1 stimuli. Long-latency (> 10 ms) EPSPs were seen in both cell types in response to presubicular, but not entorhinal or CA1 stimulation. Bursting cells responded to brief trains of orthodromic stimuli (2-10 pulses, 5-10 ms interstimulus interval) with a burst of action potentials even when the cell was previously depolarized out of bursting range by current injection. Non-bursting cells responded to brief trains of orthodromic stimuli with repetitive firing (< or = 1 spike/stimulus) at all holding potentials. Spike intervals could reach those seen in bursts by bursting cells. It is concluded that: (1) the distinction between bursting and non-bursting subicular neurons is a dichotomy and cells do not change their identity when activated antidromically or orthodromically; (2) the outputs of the two cell types may be different: bursting cells projected to presubiculum and non-bursting cells projected to entorhinal cortex; and (3) non-bursting cells can, when repetitively stimulated, fire repetitive spikes with interspike intervals in the range of intervals seen in bursts.

Action Potentials↗
PubMed1997

Proviral insertions induce the expression of bone-specific isoforms of PEBP2alphaA (CBFA1): evidence for a new myc collaborating oncogene.

The til-1 locus was identified as a common retroviral integration site in virus-accelerated lymphomas of CD2-myc transgenic mice. We now show that viral insertions at til-1 lead to transcriptional activation of PEBP2alphaA (CBFA1), a transcription factor related to the Drosophila segmentation gene product, Runt. Insertions are upstream and in the opposite orientation to the gene and appear to activate a variant promoter that is normally silent in T cells. Activity of this promoter was detected in rodent osteogenic sarcoma cells and primary osteoblasts, implicating bone as the normal site of promoter activity. The isoforms encoded by the activated gene all encompass the conserved runt DNA-binding domain and share a novel N terminus different from the previously reported PEBP2alphaA products. Minor products include isoforms with internal deletions due to exon skipping and a novel C-terminal domain unrelated to known runt domain factors. The major isoform expressed from the activated til-1 locus (G1) was found to account for virtually all of the core binding factor activity in nuclear extracts from its corresponding lymphoma cell line. Another member of this gene family, AML1(CBFA2), is well known for its involvement in human hemopoietic tumors. These results provide evidence of a direct oncogenic role for PEBP2alphaA and indicate that the Myc and Runt family genes can cooperate in oncogenesis.

Amino Acid Sequence↗
PubMed1997

Streamwater nitrate concentrations in six agricultural catchments in Scotland.

The concentrations of nitrate-N (NO3-N) in catchment inputs and outputs have been compared and contrasted between 6 farm catchments in Scotland, 3 in the West and 3 in the North-East. Forms of intensive animal farming ranging between beef and dairy cattle, sheep and poultry give different sources for potential NO3-N leakage from the systems. While stream reaches bordered by intensive cereal production give rise to the largest inputs to surface waters, climatic influences result in the more-efficient use of fertilizer- and farm waste-N in the West, and an enhanced potential for N-loss to waters in the cooler North-East, regardless of the N-inputs being considerably lower in the latter region. Although the EC Nitrate Directive limit of 11.3 mg NO3-N 1(-1) was not exceeded, peak values occurring during summer baseflows and autumn soil rewetting were commonly larger than the 'target' maximum concentration of 5.65 mg NO3-N 1-1.

Agriculture↗
PubMed1997

Presubicular and parasubicular cortical neurons of the rat: functional separation of deep and superficial neurons in vitro.

1. The presubiculum and parasubiculum are retrohippocampal structures bordered by the subiculum and medial entorhinal cortex. Deep layer (IV-VI) neurons from this region exhibit stable synaptically triggered burst behaviour which distinguishes them from superficial layer (I-III) cells. This functional separation was examined with intracellular and field potential recordings from horizontal slices of rat brain. Neurobiotin labelling and rapid Golgi techniques were used to obtain anatomical evidence of axonal trajectories. 2. Extracellular stimulation of the subiculum, deep medial entorhinal cortex or superficial pre- or parasubiculum caused, in deep layer cells only, a short latency burst discharge which could be followed by one or more after-discharges. Bursts appeared after repetitive stimulation and were stable for the life of the slice. Each event was supported by giant excitatory postsynaptic potentials (EPSPs). Events were similar whether they were evoked in horizontal slices or slices cut perpendicular to the horizontal plane. 3. Bath application of the NMDA receptor antagonist 3-[2-carboxypiperazin-4-yl]-propyl-1-phosphonic acid (CPP; 5 microM) elevated the threshold for evoking the giant EPSP. Higher concentrations (10-15 microM) reduced the amplitude and duration of the giant EPSP. Bath application of the AMPA receptor antagonist beta-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 5 microM) eliminated the evoked EPSP. 4. In intact slices, superficial layer neurons of pre- and parasubiculum could exhibit EPSPs coincident with bursts recorded in the deep layers. However, in isolated subsections of horizontal slices or in 'vertical slices', both of which contained only pre- and/or parasubiculum, evoked or picrotoxin-induced bursts occurred only in deep layer cells. Superficial layer cells in these subsections showed no response to deep layer events. 5. Antidromic population spikes confirmed projections from superficial cell layers of pre- and parasubiculum down to their deep cell layers. Reciprocal antidromic responses were absent. 6. Axons of superficial layer stellate and pyramidal cells had horizontal collaterals and at least one ascending and one descending collateral. Branches of the descending collaterals were given off in layer V and some axons were found to reach the angular bundle. Axons of deep layer stellate and pyramidal cells also had horizontal collaterals and descending collaterals which could be traced to the angular bundle. One ascending axon collateral was found among the thirty-one deep layer cells examined morphologically. 7. We conclude that the deep layer cells of the presubiculum and parasubiculum are richly interconnected with excitatory synapses. These interconnections can generate giant excitatory synaptic potentials that support the bursting behaviour exhibited by these neurons. Any of the excitatory inputs to deep layer cells can trigger the population bursts and specific inputs from entorhinal cortex produce the after-discharges. Further, connections between superficial and deep layer cells appear to be almost exclusively in the direction of superficial to deep. The absence of significant ascending input can account for the functional separation of superficial and deep layer neurons of presubiculum and parasubiculum.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1997

Molecular interactions between the importin alpha/beta heterodimer and proteins involved in vertebrate nuclear protein import.

We have used in vitro binding assays to examine specific interactions between a number of cytoplasmic and nuclear pore proteins involved in nuclear protein import in vertebrates. We demonstrate that nuclear transport factor 2 (NTF2), nucleoporin p62 and the Ras-like GTPase Ran bind to the importin heterodimer via its beta subunit. The binding behaviour of p62 truncation mutants indicated that importin-beta interacts primarily with the alpha-helical coiled-coil rod domain of nucleoporin p62 and not with the N-terminal domain that contains a number of degenerate repeats based on the xFxFG sequence motif. The binding of Ran to importin-beta was sensitive to its nucleotide state, with RanGTP binding strongly, whereas RanGDP binding could not be detected using our assay conditions. RanGTP, but not RanGDP, was able to displace p62 bound to the importin alpha/beta complex, suggesting that the binding sites for p62 and RanGTP on importin-beta overlap. Moreover, RanGTP, but not RanGDP, weakened the interaction between importin-alpha and importin-beta in a concentration-dependent manner. NTF2 bound to the importin heterodimer but did not displace p62, suggesting that the NTF2 and p62 binding sites on importin-beta do not overlap. The set of interactions we observed was not altered by the binding of NLS-containing substrates such as transcription factor IIIA to the importin heterodimer. Our results are consistent with models for nuclear protein import in which Ran nucleotide exchange modulates the binding of the importin-substrate complexes during translocation through nuclear pore complexes.

Amino Acid Sequence↗
PubMed1997

Presubicular and parasubicular cortical neurons of the rat: electrophysiological and morphological properties.

Intracellular recordings and Neurobiotin-injection were used to examine the electrophysiology and morphology of presubicular and parasubicular cortical neurons in horizontal slices from rat brains. Evoked responses were obtained by stimulation of subicular and entorhinal cortices. Stellate cells were recorded in layers II and V of presubiculum and parasubiculum. Superficial layer cells had spiny dendrites that were found to reach layer I. Deep layer cells had sparsely spiny dendrites or dendrites without spines that did not reach past layer IV. Pyramidal cells were recorded in layers III and V of presubiculum and layers II and V of parasubiculum. Superficial layer cells had spiny dendrites that were found to reach layer I. Deep layer cells had sparsely spiny dendrites or dendrites without spines that could reach layer II. Electrophysiologically, stellate and pyramidal cells were similar to one another, regardless of cell layer, exhibiting repetitive single spiking in response to depolarizing current injection. No cells were found to burst in response to current injection. While there were subtle electrophysiological differences among the cell types, stellate cells were more similar to pyramidal cells from the same or adjacent layers than to other stellate cells from more distant layers. Similarly, pyramidal cells were electrophysiologically more similar to nearby stellate cells than to other distant pyramidal cells. Cells of all layers responded to subicular stimulation with a short latency (< 9 ms), excitatory postsynaptic potential. Superficial layer cells responded at short (< 9 ms), longer (10-20 ms) and very long latencies (> 20 ms) to stimulation of superficial layers of medial entorhinal cortex. Deep layer cells responded at short latencies (< 9 ms) to stimulation of deep layers of medial entorhinal cortex. Many cells responded to both subicular and entorhinal inputs. Both pyramidal and stellate cells in the deep layer of pre/parasubiculum could exhibit population bursting behavior in response to stimulation of subiculum or entorhinal cortex. The results define the cellular morphology and basic electrophysiology of presubicular and parasubicular neurons of the rat brain as a step toward understanding the physiology of the retrohippocampal cortices.

Action Potentials↗
PubMed1997

Crystallization and preliminary X-Ray diffraction characterization of a dimerizing fragment of the rod domain of the Dictyostelium gelation factor (ABP-120).

We have expressed in Escherichia coli a construct corresponding to sequence repeats 5 and 6 of the rod domain of the actin-binding protein Dictyostelium gelation factor (ABP-120). We have obtained orthorhombic P212121 crystals of the protein with a = 43.5 A, b = 103.2 A, c = 124.4 A. These crystals diffract past 2.2 A resolution using synchrotron radiation and are suitable for high-resolution structural analysis. ABP-120 is a key component of the Dictyostelium cytoskeleton, where it functions to crosslink F-actin filaments into networks. This crosslinking function of ABP-120 depends crucially on the formation of dimeric molecules that contain an actin-binding site on each chain, and this dimerization is brought about through interactions between repeating sequence modules in the rod domain. Because the construct we have expressed retains the ability to dimerize, it should enable us to establish the precise manner in which these sequence repeats interact with one another in the intact molecule.

Animals↗
PubMed1997

Support and coping of male hemodialysis-dependent patients.

The purpose of this descriptive-correlational study was to describe coping strategies used by males with chronic renal failure who are dependent on hemodialysis; to describe their social networks; to describe the perceived support, conflict, and reciprocity within their interpersonal relationships; and to examine the relationships among the variables support, conflict, reciprocity, social networks, and coping strategies. Social support was conceptualized as a coping resource or source of assistance in coping with the renal illness- or hemodialysis-related stressor. The Ways of Coping questionnaire and the Interpersonal Relationship Inventory were administered to 30 participants while in hospital. Although, both problem-focused and emotion-focused forms of coping were used, participants primarily used problem-focused coping, in particular, "seeking social support". Overall, the participants perceived relatively high levels of support and moderate to high levels of reciprocity with members of their social networks. Participants experienced a moderate level of conflict in their interpersonal relationships. Both escape-avoidance and conflict were positively associated with the number of people in the household. Positive reappraisal was negatively associated with the number of close relatives. The small sample size prohibits generalizability of the results. Longitudinal studies with a larger randomly selected sample would yield insights into the long-term psychological outcomes of different coping strategies and into the bi-directional relationship of support from social network and coping in this population. Implications for nurses are discussed.

Adaptation, Psychological↗
PubMed1997

The effect of growth hormone replacement therapy for up to 12 months on lipoprotein composition and lipoprotein(a) in growth hormone-deficient adults.

The effect of growth hormone replacement therapy in near physiological doses on lipoprotein composition and serum lipoprotein(a) concentrations was investigated in growth hormone-deficient subjects. A randomised, double-blind, placebo-controlled trial of recombinant growth hormone was undertaken for 6 months followed by an open extension for a further 6 months (0.125 IU/kg per week for the first 4 weeks of each 6 month period and thereafter 0.25 IU/kg per week). A total of 18 patients with isolated growth hormone deficiency or hypopituitarism were studied. Lipid concentrations were estimated in lipoprotein fractions and protein concentrations were measured in low density lipoprotein (LDL). Glucose and glycated haemoglobin in blood and insulin, cholesterol, triglyceride, apolipoproteins A-I and B and lipoprotein(a) concentrations were measured in serum. In the placebo-controlled phase fasting blood glucose concentrations increased with growth hormone treatment from 5.0 +/- 0.2 to 5.8 +/- 0.2 mmol/l (P = 0.02) (mean +/- S.E.M.), although no significant changes were seen in lipids or lipoproteins. In the group receiving active treatment total serum cholesterol decreased from 6.0 +/- 0.4 to 5.2 +/- 0.3 mmol/l (P = 0.002) after 6 months, due to reduced LDL cholesterol concentrations. Low density lipoprotein protein concentrations fell (0.8 +/- 0.1 versus 0.7 +/- 0.1 g/l) (P = 0.005), and LDL phospholipid levels decreased from 0.9 +/- 0.1 to 0.7 +/- 0.1 mmol/l (P = 0.007). Serum cholesterol and LDL composition reverted to pre-treatment values by 12 months. Fasting blood glucose remained above pre-treatment values (P = 0.036) and fasting insulin was significantly increased (P = 0.044). There was no effect of growth hormone therapy on serum triglyceride, apolipoprotein or lipoprotein(a) concentrations. In conclusion, growth hormone therapy with near physiological doses has no long term effects on serum lipoprotein(a) concentrations or lipoprotein composition.

Adult↗
PubMed1997

Orbital involvement in allergic fungal sinusitis.

BACKGROUND: Although allergic fungal sinusitis is a relatively common, noninvasive form of paranasal sinus mycosis, and despite frequent orbital involvement, there have been few reports of this condition in the ophthalmic literature. METHODS: Two cases of allergic fungal sinusitis having orbital symptoms are described. The current classification, typical presentation, and ideal management of fungal sinusitis are reviewed. RESULTS: Distinguishing radiologic and pathologic features were present in both patients. Aspergillus flavus was cultured in one case, and Bipolaris spicifera was cultured in the other. CONCLUSIONS: Allergic fungal sinusitis is a unique subset of sino-orbital disease with highly characteristic clinical, radiologic, and pathologic features. Unlike invasive forms of mycotic disease, allergic fungal sinusitis may be managed adequately with surgical debridement, aeration of the involved sinuses, and systemic and topical corticosteroids.

Adult↗
PubMed1997

Fostering children's resilience.

Resilience is relevant to nurses because of its implications for health. Research on the resilience of children and adolescents has proliferated over the past five years. However, the specific processes underlying resilience and outcome variables require further study. Furthermore, few intervention studies have been conducted. This article describes resilience and factors that influence resilience of children, examines the relationship between resilience and health, identifies interventions that foster children's resilience and health, reviews research focusing on children's resilience, and suggests the relevance of resilience to nursing of children.

Adaptation, Psychological↗
PubMed1997

Pharmacokinetics and safety of a single dose of stavudine (d4T) in patients with severe hepatic impairment.

This open-label study enrolled five subjects with biopsy-proven cirrhosis and moderate to severe hepatic impairment (Child-Pugh classification grade B or C) and five age- and gender-matched controls. All subjects received a single 40-mg oral dose of stavudine (d4T). Stavudine pharmacokinetics in subjects with hepatic impairment were similar to those in age- and gender-matched control subjects and were not substantially different from those previously observed in human immunodeficiency virus-infected patients. Based on these findings, stavudine use does not require modification of the dose or dosing interval for patients with liver disease.

Administration, Oral↗
PubMed1997

Interaction between the small GTPase Ran/Gsp1p and Ntf2p is required for nuclear transport.

Bidirectional movement of proteins and RNAs across the nuclear envelope requires Ran, a Ras-like GTPase. A genetic screen of the yeast Saccharomyces cerevisiae was performed to isolate conditional alleles of GSP1, a gene that encodes a homolog of Ran. Two temperature-sensitive alleles, gsp1-1 and gsp1-2, were isolated. The mutations in these two alleles map to regions that are structurally conserved between different members of the Ras family. Each mutant strain exhibits various nuclear transport defects. Both biochemical and genetic experiments indicate a decreased interaction between Ntf2p, a factor which is required for protein import, and the mutant GSP1 gene products. Overexpression of NTF2 can suppress the temperature sensitive phenotype of gsp1-1 and gsp1-2 and partially rescue nuclear transport defects. However, overexpression of a mutant allele of NTF2 with decreased binding to Gsp1p cannot rescue the temperature sensitivity of gsp1-1 and gsp1-2. Taken together, these data demonstrate that the interaction between Gsp1p and Ntf2p is critical for nuclear transport.

Amino Acid Sequence↗
PubMed1997

Fragile X, iron, and neurodevelopmental screening in 8 year old children with mild to moderate learning difficulties.

OBJECTIVE: To examine the value of neurodevelopmental examination, fragile X testing, iron studies, and other screening procedures in children with mild to moderate learning difficulties. DESIGN: A cross sectional case-control study. SUBJECTS: A 34% random sample (n = 130) of children with mild to moderate learning difficulties born between 01/07/83 and 30/06/84 and resident in North and West Belfast. Controls were 130 children without learning difficulties matched for age and geographical area. RESULTS: The prevalence of mild to moderate learning difficulties in North and West Belfast was 16%; 115 (89%) of the learning difficulties group and 80 (58%) of the control group consented to participate. Children in the learning difficulties group scored significantly lower in neurodevelopmental testing and were more likely to fail their audiometry assessment than controls. Children in the learning difficulties group were also more likely to be anaemic and had lower serum iron and transferrin saturation than controls. No cases of fragile X were identified. Thyroid function tests and urinary amino acids were all within normal limits There were no significant differences in anthropometry, head circumference, or formal neurological examinations. CONCLUSIONS: Children with learning difficulties are more likely to be neurodevelopmentally immature and iron depleted than controls. Iron depletion should be aggressively sought and treated. The role for routine assessment for fragile X, thyroid function tests, and amino acid chromatography is doubtful.

Amino Acids↗
PubMed1997

Calreticulin, a potential vascular regulatory protein, reduces intimal hyperplasia after arterial injury.

Both thrombotic and inflammatory responses to arterial injury have been implicated in atherosclerotic plaque growth. Calreticulin is a ubiquitous calcium-binding protein with antithrombotic activity and, in addition, is associated with leukocyte activation. We are investigating calreticulin as a potential vascular regulatory protein. The development of intimal hyperplasia was studied at sites of balloon injury in iliofemoral arteries from 91 rats. Calreticulin was infused directly into the artery immediately before balloon injury, and plaque growth was then assessed at 4 weeks' follow-up. Parallel studies of the effects of each calreticulin domain as well as a related calcium-binding protein, calsequestrin, were examined. The effects of calreticulin on platelet activation, clot formation, and mononuclear cell migration were also studied. When infused before balloon injury in rat iliofemoral arteries, calreticulin, or its high-capacity Ca(2+)-binding C domain, significantly reduces plaque development, whereas calsequestrin, a related calcium-binding protein that lacks the multifunctional nature of calreticulin, does not decrease plaque area (saline: 0.037 +/- 0.007 mm2, calsequestrin: 0.042 +/- 0.021 mm2, calreticulin: 0.003 +/- 0.002 mm2, n = 46, P < .04). The N domain and more specifically the P domain, a low-capacity, high-affinity calcium-binding domain in calreticulin, do not reduce intimal hyperplasia (N + P domain: 0.038 +/- 0.012 mm2, C domain: 0.003 +/- 0.002 mm2, n = 45 rats, P < .0001). Calreticulin reduces macrophage and T cell staining in the arterial wall after injury but has no direct effect on monocyte migration in vitro (percent medial area staining positive for macrophage 24 hours after injury (N + P: 4.06 +/- 1.42, calreticulin: 0.273 +/- 0.02; n = 26, P < .009). Calreticulin does, however, reduce platelet-dependent whole blood clotting time, in vitro (baseline: 78.23 +/- 2.04 seconds, calreticulin: 113.5 +/- 1.95 seconds; n = 5, P < .002). We conclude that calreticulin significantly reduces intimal hyperplasia after arterial injury, potentially acting as a vascular regulatory protein.

Angioplasty, Balloon↗
PubMed1997

Nipple wound care: a new approach to an old problem.

Nipple soreness and nipple trauma have long been associated with breastfeeding, and persist despite the many clinical advancements in the field of lactation. Management of nipple wounds has been approached in a variety of ways over the years in an attempt to treat and resolve this problem. Incorporating the scientific principles of modern wound care management can provide additional effective treatment options. Wounds are characterized by depth and extent of tissue destruction, regardless of their location on the body. Current wound treatment methods employ the use of moisture to aid healing. A moist environment is critical for epithelization, the proliferation and migration of epithelial cells across the surface of a wound during healing. Nipple wounds also heal by this process. Using a particular type of wound dressing, a hydrogel sheet wound covering, on a nipple wound offers several advantages. These dressings help maintain a moist environment, decrease the chance of bacterial infection, are easy to use, and provide immediate pain relief.

Breast Feeding↗