Biomedical subjects

R C Elliott

Publications and source records attributed to R C Elliott.

At least 19 recordsLinked to original sources

PubMed2000

Changes in activating protein 1 (AP-1) composition correspond with the biphasic profile of nerve growth factor mRNA expression in rat hippocampus after hilus lesion-induced seizures.

In adult brain, nerve growth factor (NGF) gene expression is generally upregulated by neuronal activity. However, a single episode of hilus lesion (HL)-induced limbic seizures stimulates a biphasic increase in NGF mRNA expression with peaks at 4-6 and 24 hr after lesion and an intervening return to control levels at 10-12 hr after lesion. In vitro studies suggest that NGF transcription is regulated via an activating protein 1 (AP-1) binding site in the first intron of the NGF gene. To examine the relationship between seizure-induced AP-1 binding and NGF gene expression in this paradigm, NGF mRNA levels and AP-1 binding were examined after HL seizures. Furthermore, to gain insight into the functional composition of the AP-1 complex, supershift analysis was performed to characterize which Fos and Jun family members are included in the AP-1-binding complex at the different time points analyzed. Solution hybridization analysis verified the biphasic increase in NGF mRNA content of the dentate gyrus after HL seizures. After an initial increase, AP-1 binding slowly declined in a stepwise manner that encompassed, but did not correspond with, the two phases of NGF mRNA expression. However, supershift analyses demonstrated that the relative contributions of JunD and JunB to the AP-1 complex exhibited positive and negative correlations, respectively, with the phases of increased NGF expression after HL. These results suggest that AP-1 complexes containing JunD promote NGF transactivation and that transient changes in the relative contributions of JunD and JunB to AP-1 binding underlie the biphasic increase in NGF gene expression induced by HL seizures.

Age Factors↗

PubMed2000

Evidence that auxin promotes gibberellin A1 biosynthesis in pea.

In shoots of the garden pea, the bioactive gibberellin (GA1) is synthesised from GA20, and the enzyme which catalyses this step (a GA 3-oxidase -- PsGA3ox1) is encoded by Mendel's LE gene. It has been reported previously that decapitation of the shoot (excision of the apical bud) dramatically reduces the conversion of [3H]GA20 to [3H]GA1 in stems, and here we show that endogenous GA1 and PsGA3ox1 transcript levels are similarly reduced. We show also that these effects of decapitation are completely reversed by application of the auxin indole-3-acetic acid (IAA) to the 'stump' of decapitated plants. Gibberellin A20 is also converted to an inactive product, GA29, and this step is catalysed by a GA 2-oxidase, PsGA2ox1. In contrast to PsGA3ox1, PsGA2ox1 transcript levels were increased by decapitation and reduced by IAA application. Decapitation and IAA treatment did not markedly affect the level of GA1 precursors. It is suggested that in intact pea plants, auxin from the apical bud moves into the elongating internodes where it (directly or indirectly) maintains PsGA3ox1 transcript levels and, consequently, GA1 biosynthesis.

Catalysis↗

PubMed1999

Spontaneous abortion in the British semiconductor industry: An HSE investigation. Health and Safety Executive.

BACKGROUND: The UK Health and Safety Executive (HSE) conducted a study to examine the risk of spontaneous abortion (SAB) in British female semiconductor industry workers, following reports from the USA which suggested an association between risk of SAB and work in fabrication rooms and/or exposure to ethylene glycol ethers. METHODS: A nested case-control study based on 2,207 women who had worked at eight manufacturing sites during a 5-year retrospective time frame was established; 36 cases were matched with 80 controls. RESULTS: The overall SAB rate in the industry was 10.0%. (65 SABs/651 pregnancies) The crude odds ratio (OR) for fabrication work was 0.65 (95% CI 0.30-1.40). This was essentially unchanged after adjustment for a range of potential confounding factors in the first 3 months of pregnancy and was reduced to 0.58 (95% CI 0.26-1.30) after adjustment for smoking in the previous 12 months. There were no statistically significantly elevated ORs for any work group or any specific chemical or physical exposure in the industry. CONCLUSIONS: There is no evidence of an increased risk of SAB in the British semiconductor industry. Am. J. Ind. Med. 36:557-572, 1999. Published 1999 Wiley-Liss, Inc.

Abortion, Spontaneous↗

PubMed1999

Localization of GABA(B) (R1) receptors in the rat hippocampus by immunocytochemistry and high resolution autoradiography, with specific reference to its localization in identified hippocampal interneuron subpopulations.

Immunocytochemical and autoradiographic methods were used to localize the GABA(B) receptor in the normal rat hippocampus. GABA(B) receptor 1-like immunoreactivity (GBR1-LI) was most intense in presumed GABAergic interneurons of all hippocampal subregions. It was also present throughout the hippocampal neuropil, where it was most intense in the dendritic strata of the dentate gyrus, which are innervated by the perforant pathway and inhibitory dentate hilar cells, and in strata oriens and radiatum of area CA3. The dendritic regions of area CA1 exhibited less GBR1-LI than area CA3. GBR1-LI was detectable in the somata of CA1 pyramidal cells, but was minimal or undetectable within the somata of dentate granule cells and CA3 pyramidal cells. GBR1-LI was similarly minimal in the dentate hilar neuropil, and in stratum lucidum, the two regions that contain granule cell axons and terminals. Nor was GBR1-LI detectable in the inhibitory basket cell fiber systems that surround hippocampal principal cell somata. Fluorescence co-localization studies indicated that significant proportions of interneurons expressing somatostatin, neuropeptide Y, cholecystokinin, calbindin, or calretinin also expressed GBR1-LI constitutively. Conversely, parvalbumin-positive GABAergic basket cells of the dentate gyrus and hippocampus, which form GABA(A) receptor-mediated inhibitory axo-somatic synapses, rarely contained detectable GBR1-LI. High resolution autoradiography with the GABA(B) receptor antagonist CGP 62349 revealed a close correspondence between receptor ligand binding and GBR1-LI, with several notable exceptions. Ligand binding closely matched GBR1-LI throughout the hippocampal, cortical, thalamic, and cerebellar neuropil. However, the hippocampal interneuron somata and dendrites that exhibited the most intense GBR1-LI, and the GBR1-positive somata of CA1 pyramidal cells, did not exhibit a similar density of [3H]-CGP 62349 binding. These data clarify the relationship between immunocytochemically identified receptor protein and potentially functional receptors, indicating that GBR1-LI reflects both non-functional cytoplasmic GBR1 and the ligand-bindable form of the protein, both before dimerization with GBR2 and after translocation to functional sites within cells. The staining and binding patterns further suggest that GBR1 is constitutively expressed in specific neuronal populations, and may exist in higher concentration in the axons of inhibitory hippocampal pathways that innervate dendritic zones, than in axo-somatic inhibitory terminals. Whether GBR1 is inducible in cells that contain GBR1 mRNA, but no detectable constitutive protein, remains to be determined in experimental studies.

Animals↗

PubMed1999

Gibberellin 2-oxidation and the SLN gene of Pisum sativum.

Two cDNAs encoding gibberellin 2-oxidases were isolated from maturing pea seeds. The first, PsGA2ox1, was isolated by activity screening of a Lambda-ZAP cDNA library excised into phagemid form and expressed in Escherichia coli. The second, PsGA2ox2, was obtained initially as a PCR product using degenerate primers designed according to conserved regions of plant 2-oxoglutarate-dependent dioxygenases. E. coli heterologous expression products of PsGA2ox1 and PsGA2ox2 converted GA1 to GA8, as shown by HPLC-radiocounting, and gas chromatography-MS. PsGA2ox1 converted GA20 to GA29, but GA20 was a poor substrate for the PsGA2ox2 expression product. Furthermore, PsGA2ox1 converted GA29 to GA29-catabolite at a low level of efficiency while PsGA2ox2 did not catalyse this step. A cDNA of PsGA2ox1 isolated from plants of genotype sln contained a single base deletion which was predicted to produce a truncated protein and gibberellin 2-oxidase activity could not be demonstrated from this cDNA. A 10 bp size difference between the introns of the SLN and sln PsGA2ox1 genes was used to show co-segregation between the SLN and sln phenotypes and the size of the PCR products. PsGA2ox1 transcripts were more abundant in cotyledons than in shoots, while the reverse was the case for PsGA2ox2. The expression patterns of the genes, together with the effects of the sln mutation, indicate that PsGA2ox1 plays a major role in GA20 deactivation in both shoots and maturing seeds, while the PsGA2ox2 gene might be important for GA1 deactivation in the shoot.

Amino Acid Sequence↗

PubMed1998

Unilateral hippocampal lesions in newborn and adult rats: effects on spatial memory and BDNF gene expression.

Subcortical damage at birth often produces more severe deficits than similar lesions in an adult. In the present study, effects of unilateral electrolytic hippocampal ablations made on postnatal day 1 or in 3-month-old adult rats, were compared. Exploratory behavior and spatial navigation in the Morris water maze (MWM) were assessed 8 and 20 weeks after hippocampal damage. Rats with neonatal damage did not respond to novelty in the environment and did not learn to find the hidden platform in the MWM. Rats lesioned as adults did learn the water maze task, but slower than controls. We hypothesized that behavioral deficits observed in rats lesioned at birth, may be due, in part, to neurochemical dysfunction of the contralateral hippocampus. Specifically, cholinergic and GABAergic neurotransmission were assessed by measuring choline-acetyltransferase (ChAT) and GABAdecarboxylase (GAD) activity. In addition, nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) mRNA levels were assayed in the remaining (contralateral) hippocampus. Of these molecules, only BDNF gene expression was significantly reduced (by 30%) at 8 and 20 weeks after neonatal and adult unilateral ablation. The similar reduction in BDNF mRNA in both treatment groups does not correspond with the lesion's differential effect on memory function. However, the more severe learning impairment after neonatal lesion may reflect increased dependence on trophins during development.

Aging↗

PubMed1996

The coronary vasoconstrictor action of extract IV from the dahlia sea anemone Tealia felina L.

Extract IV, a partially purified preparation of the toxin from the sea anemone Tealia felina, produced marked bradycardia and arrhythmias in the rat in vivo. In the Langendorff rat heart preparation perfused at constant pressure extract IV (0.0224 AU/ml) reduced the force of contraction by 81.3 +/- 7.2%, n = 6, and the coronary flow by 82.0 +/- 4.7%, n = 5. When the preparation was perfused at constant flow rate, extract IV (0.072 AU/ml) increased the coronary circulation resistance from 3.76 +/- 1.09 (control) to 36.94 +/- 12.26 mmHg/ml/min. The force of contraction was also markedly reduced. The extract produced bradycardia in preparations perfused at constant pressure but not in those perfused at constant flow rate. The extract did not affect isolated atria preparations. It was concluded that the bradycardia produced in vivo and in preparations perfused in vitro at constant pressure was probably secondary to the coronary vasospasm produced, which could contribute to the cardiotoxicity of the extract.

Animals↗

PubMed1996

AINTEGUMENTA, an APETALA2-like gene of Arabidopsis with pleiotropic roles in ovule development and floral organ growth.

To understand better the role of genes in controlling ovule development, a female-sterile mutant, aintegumenta (ant), was isolated from Arabidopsis. In ovules of this mutant, integuments do not develop and megasporogenesis is blocked at the tetrad stage. As a pleiotropic effect, narrower floral organs arise in reduced numbers. More complete loss of floral organs occurs when the ant mutant is combined with the floral homeotic mutant apetala2, suggesting that the two genes share functions in initiating floral organ development. The ANT gene was cloned by transposon tagging, and sequence analysis showed that it is a member of the APETALA2-like family of transcription factor genes. The expression pattern of ANT in floral and vegetative tissues indicates that it is involved not only in the initiation of integuments but also in the initiation and early growth of all primorida except roots.

Amino Acid Sequence↗

PubMed1995

NGF and BDNF are differentially modulated by visual experience in the developing geniculocortical pathway.

Neuronal activity and trophic factors have been implicated in shaping the connectivity of functional synaptic circuits. We studied the development and regulation by sensory input of the neurotrophins NGF, BDNF and NT-3 in the developing rat visual system. In the occipital cortex, NT-3 mRNA was transiently expressed in the neonate. In contrast, BDNF and NGF mRNA's increased during postnatal development, and reached mature levels around 3 weeks of age. BDNF mRNA was ten times more abundant than NGF mRNA. In the lateral geniculate nucleus (LGN), NT-3 mRNA was also transiently expressed, whereas NGF and BDNF mRNA's did not vary significantly during development. The high-affinity neurotrophin receptors trkB and trkC were expressed both in the developing LGN and occipital cortex. These receptors for BDNF and NT-3, respectively, were expressed at birth, with little change during development. In contrast, trkA mRNA, which encodes the high-affinity NGF receptor, was undetectable in either region. Visual experience differentially modulated expression of NGF and BDNF mRNA's. NGF mRNA was slightly increased after 3 weeks of light-deprivation. In contrast, BDNF mRNA expression in visual cortex was significantly lower than normal in rats dark-reared from birth. Decreased BDNF expression after sensory deprivation was reversible by exposure to light. Thus, all three neurotrophins were detected in visual cortex and LGN. Differences in abundance developmental profiles, and regulation imply distinct functions for each factor in the visual system.

Aging↗

PubMed1994

An improved method detects differential NGF and BDNF gene expression in response to depolarization in cultured hippocampal neurons.

Differential regulation of individual neurotrophins by impulse activity potentially allows transformation of instantaneous signalling into diverse, long-lasting neural alterations. To define the temporal profiles of trophin gene expression we examined nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) mRNAs in dissociated cell cultures of rat hippocampus using an improved solution hybridization technique. Traditional methods lack the precision and sensitivity to detect small changes during brief intervals and the facility to process large sample numbers simultaneously. This improved method has now allowed us to better define the dynamics of depolarization-induced changes in expression of individual trophin genes. Using elevated K+ as a depolarizing stimulus, NGF mRNA increased 40% after 48 h. In contrast, BDNF message rose almost 4-fold within 3 h and attained a maximal 6-fold increase within 6 h. Similar increases in BDNF mRNA levels were exhibited following treatment of cultures with glutamate, an excitatory neurotransmitter. To document the sensitivity of BDNF mRNA to depolarizing conditions, we examined expression after K+ withdrawal. BDNF message began decreasing within one hour post-depolarization, and returned to basal levels after 6 h. Observations indicate that BDNF and NGF mRNAs are induced differentially in response to impulse activity; BDNF message is acutely responsive to ongoing changes, whereas NGF mRNA responds more slowly and sluggishly. The physiological implications of this differential regulation are discussed.

Animals↗

PubMed1993

Depolarizing influences increase low-affinity NGF receptor gene expression in cultured Purkinje neurons.

Multiple cellular and molecular interactions are required for the differentiation and development of central neurons. For example, neural activity may modulate trophic function. In the developing cerebellum, establishment of functional excitatory synaptic connections coincides with the expression of NGF and its receptors. We have previously shown that excitatory signals and NGF act in concert to regulate the survival and morphological differentiation of cerebellar Purkinje cells in culture. To begin investigating the molecular mechanisms by which trophic interactions and neural activity modulate cerebellar development, we have now studied the role of excitatory signals on the expression of both NGF and the p75 glycoprotein (the low-affinity component of the NGF receptor) by cerebellar cells in culture. We used p75 as a model of potential responsiveness, since it is well characterized and conveniently monitored. Expression of the NGF and p75 mRNA's was studied in either mixed, neuron-enriched, or pure glial cultures. Expression of the NGF gene was localized to proliferating glial cells, while expression of p75 was restricted to developing Purkinje cells. To evaluate whether presynaptic activation may potentially modulate trophic factor receptor expression, the expression of the p75 gene was studied in cultures exposed to excitatory signals. Depolarization of cultures with high potassium, veratridine, or exposure to the excitatory neurotransmitter aspartate, resulted in a two- to threefold increase in the expression of both the p75 protein and messenger RNA. These increases did not require the presence of glia, indicating a direct effect of the excitatory signals on the neuronal population. Moreover, message and receptor increased per neuron. Our study suggests that local glia provide trophic support for Purkinje cell development, and that impulse activity modulates Purkinje cell responsiveness by regulating expression of trophic receptor subunits.

Animals↗

PubMed1992

Initial characterization of a pea mutant with light-independent photomorphogenesis.

We have identified a mutant of pea cultivar Alaska that has many of the characteristics normally associated with light-grown seedlings even when grown in complete darkness. We have designated this mutant lip1, for light independent photomorphogenesis. Etiolated wild-type pea seedlings are white to slightly yellow in color and have a distinct morphology characterized by elongated epicotyls and buds containing unexpanded leaves with small, undifferentiated cells. In contrast, mutant seedlings grown under the same conditions are yellow in color and have short epicotyls and expanded leaves showing clear cellular differentiation. Transmission electron microscopy revealed partially developed, agranal plastids in the dark-grown mutant, unlike wild-type seedlings that contain etioplasts with prolamellar bodies. The mutant also exhibits a much shorter lag period for chlorophyll accumulation when etiolated seedlings are transferred from darkness to white light. The dark-grown mutant has 10-fold less spectrally detectable phytochrome, which can be attributed to a 10-fold reduction in the level of the PHYA polypeptide. Cab, Fed1, and RbcS transcripts are present in dark-grown mutant seedlings at levels comparable to those produced in light-grown material. The levels of these transcripts show a normal decrease when green plants grown for 15 days in a light/dark cycle are transferred to continuous darkness. However, transcript levels remain high during dark treatment of seedlings grown for 9 days in continuous light, indicating that the dark adaptation response in this mutant is developmentally plastic. The lip1 mutant has several features in common with the deetiolated Arabidopsis mutants det1, det2, and cop1. However, there are also several important differences, including varying effects on phytochrome levels, organ-specific gene expression, plastid development, and response to dark adaptation.

Cytokinins↗

PubMed1992

Both internal and external regulatory elements control expression of the pea Fed-1 gene in transgenic tobacco seedlings.

In previous studies using leaves of light-grown transgenic tobacco plants, we have shown that sequences located within the transcribed region of the pea Fed-1 gene (encoding ferredoxin I) are major cis-acting determinants of light-regulated mRNA accumulation. However, we show here that these internal sequences are less important for the Fed-1 light response in etiolated tobacco seedlings than they are in green leaves and that upstream elements confer organ specificity and contribute significantly to Fed-1 light responses in etiolated material. Light effects mediated by upstream response elements are thus most pronounced during the initial induction of gene activity, whereas internal elements play a more prominent role in modulating Fed-1 expression once the gene is already active.

Base Sequence↗

PubMed1990

Role of mediastinoscopy in the diagnosis of mediastinal masses.

In order to assess the diagnostic efficiency of mediastinoscopy in patients with undiagnosed mediastinal masses, a retrospective review was undertaken of the results obtained in a consecutive series of 42 patients presenting over a 5-year period. In 28 (67%) patients, a tissue diagnosis was made. Ten (24%) patients underwent a negative mediastinoscopy but were found to have significant pathology on further investigation (eight in a location outside the range of conventional mediastinoscopy). Four (10%) patients had negative findings and were confirmed to have no significant pathology on further investigation and follow-up. Overall, mediastinoscopy had a sensitivity of 74%, a specificity of 100% and an accuracy of 76%; however, for lesions within the range of mediastinoscopy, the sensitivity was 93%, the specificity 100% and the accuracy 94%. The predictive value of a negative examination was 66%. Mediastinoscopy is a valuable technique in the diagnosis of mediastinal masses but, because of the low predictive value of a negative examination, further evaluation, including thoracotomy if necessary, is still indicated if mediastinoscopy is unhelpful.

Adolescent↗

PubMed1989

Characterization of a single copy gene encoding ferredoxin I from pea.

We have isolated, mapped, and sequenced a genomic clone containing the ferredoxin I (Fed-1) gene from Pisum sativum. The gene is present as a single copy per haploid genome. It has no introns, and it specifies a 753-nucleotide transcript encoding a 149-amino acid protein including a 52-residue transit peptide. Upstream sequences from Fed-1 contain several elements with similarity to transcriptional regulatory elements from RbcS and Cab genes, and gel mobility shift assays show that nuclear extracts from light-grown pea leaves contain one or more DNA binding activities specific for Fed-1 5'-flanking sequences. RbcS and Cab regulatory sequences are only weak competitors for this binding, however, and the RbcS and Cab similarities mostly lie outside of the region essential for binding. These data are discussed in terms of previously observed physiological differences between the light responses of Fed-1 and other genes.

Amino Acid Sequence↗

PubMed1987

Specificity of cotranslational amino-terminal processing of proteins in yeast.

Polypeptides synthesized in the cytoplasm of eukaryotes are generally initiated with methionine, but N-terminal methionine is absent from most mature proteins. Many proteins are also N alpha-acetylated. The removal of N-terminal methionine and N alpha-acetylation are catalyzed by two enzymes during translation. The substrate preferences of the methionine aminopeptidase (EC 3.4.11.x) and N alpha-acetyltransferase (EC 2.3.1.x) have been partially inferred from the distribution of amino-terminal residues and/or mutations found for appropriate mature proteins, but with some contradictions. In this study, a synthetic gene corresponding to the mature amino acid sequence of the plant protein thaumatin, expressed in yeast as a nonexported protein, i.e., lacking a signal peptide, has been used to delineate the specificities of these enzymes with respect to the penultimate amino acid. Site-directed mutagenesis, employing synthetic oligonucleotides, was utilized to construct genes encoding each of the 20 amino acids following the initiation methionine codon, and each protein derivative was isolated and characterized with respect to its amino-terminal structure. All four possible N-terminal variants--those with and without methionine and those with and without N alpha-acetylation--were obtained. These results define the specificity of these enzymes in situ and suggest that the nature of the penultimate amino-terminal residue is the major determinant of their selectivity.

Acetyltransferases↗

PubMed1987

A reappraisal of the evidence for a direct action of tetraethylammonium on the chick biventer cervicis muscle equilibrated with lidocaine.

Contractures of the chick biventer cervicis muscle were recorded in vitro in response to exogenous acetylcholine (ACh) and to tetraethylammonium (TEA) and concentration response curves (CRC) produced for these drugs. In BVC muscles equilibrated with lidocaine (1.07 mM), physostigmine (3.63 microM) blocked TEA induced contractures in a reversible non-competitive manner. It had a similar action on ACh induced contractures. Lidocaine (1.07 mM) had no action on the ACh CRC except that it reduced the response to the highest concentration of ACh tested. The data of this and of previous experiments was analyzed and a theory proposed to account for TEA contractures. It suggested that background extravesicular ACh release results in the passage of K+ into the transverse tubular system (TT). In the presence of TEA blocking the K+ channels accumulation of K+ in the TT no longer occurs. Na+/K+ exchange is depressed in favour of Na+/Ca+ exchange and the resulting rise in intracellular Ca2+ acting on the sarcoplasmic reticulum leads to contraction. Lidocaine potentiates the contraction by blocking Ca2+ uptake into the sarcoplasmic reticulum.

Acetylcholine↗

PubMed1987

The role of acetylcholine in tetraethylammonium induced contractures of the chick biventer cervicis muscle in the presence of lidocaine.

Lidocaine (0.92 mM) potentiated tetraethylammonium (TEA, 0.6-6 mM) induced contractures of the chick biventer cervicis muscle (BVC) in vitro. The dose ratio for TEA (EC50 in lidocaine/EC50 control) was 1.16 X 10(-3). Lidocaine (0.92 mM) blocked nerve and muscle action potentials in the BVC preparation, blocked slow fibre MEPPS, and blocked indirectly elicited muscle contraction. Lidocaine (0.92 mM) non-competitively blocked acetylcholine (ACh) induced contractures of the chick BVC. Gallamine competitively blocked ACh contractures (pA2 6.53) but produced only a relatively weak non-competitive block of TEA induced contractures in lidocaine. TEA induced contractures of the chick BVC in the presence of lidocaine probably do not involve ACh.

Acetylcholine↗