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Biomedical subjects

D Jordan

Publications and source records attributed to D Jordan.

At least 73 records · Page 4Linked to original sources
PubMed1995

Translation of the human papillomavirus type 16 E7 oncoprotein from bicistronic mRNA is independent of splicing events within the E6 open reading frame.

In this study we investigated the translational capacities of bicistronic and spliced mRNAs originating from the E6 and E7 regions of the high-risk genital human papillomavirus type 16 (HPV-16) and the low-risk HPV-11. For HPV-16 it was found, unexpectedly, that E7 protein could be translated from full-length bicistronic E6-E7 mRNAs. E6*I and E6*II splicing events were not required for E7 synthesis, nor did splicing increase the efficiency of E7 translation significantly. In cells, E7 synthesis from all known naturally occurring mRNA structures was very inefficient compared with that from synthetic monocistronic controls, suggesting that HPV-16 employs translational mechanisms to restrict E7 protein levels. For HPV-11, only RNAs initiated at the P264 promoter, located within the E6 open reading frame, were capable of providing an efficient template for E7 synthesis. P264-initiated mRNAs were as efficient in vivo as monocistronic controls, suggesting that the low-risk HPV-11 does not limit E7 synthesis by translational mechanisms. A detailed analysis of HPV-16 templates by using site-directed mutagenesis showed that the majority of ribosomes which ultimately translate E7 have not reinitiated after translating some or all of the upstream open reading frames. The data support a model in which the failure of 40S ribosomal initiation complexes to recognize the E6 AUG renders them capable of proceeding efficiently to translate E7.

Animals↗
PubMed1994

Quantitative autoradiography of 5-HT1D and 5-HT1E binding sites labelled by [3H]5-HT, in frontal cortex and the hippocampal region of the human brain.

In human cortex and hippocampus area, [3H]5-HT (5 nM) labels 5-HT1A, 5-HT1D and 5-HT1E sites. After masking 5-HT1A receptors by 0.1 microM 8-OH-DPAT, the binding displaced by 0.1 microM 5-CT presumably represented 5-HT1D sites and the remaining binding 5-HT1E sites. In frontal cortex, 5-HT1A receptors represented the main binding in layers II and VI and a lower fraction in other layers. 5-HT1D and 5-HT1E sites, were more homogeneously distributed in layers II to VI (21-34% of specific [3H]5-HT binding). 5-HT1E sites were of similar affinities (KD close to 6-8 nM) in the cortical layers II to VI. In CA1 field of hippocampus, (pyramidal layer, stratum radiatum, molecular layer), CA2 and dentate gyrus, 5-HT1A receptors represented the major fraction, 5-HT1D sites a significant fraction and 5-HT1E a minor fraction of the specific [3H]5-HT binding. In CA3-CA4 fields, 5-HT1A receptors were less densely present, 5-HT1D sites were predominant and 5-HT1E sites represented a significant fraction (27%). The highest densities of 5-HT1E sites have been measured in subiculum, where 5-HT1A, 5-HT1D and 5-HT1E binding sites were equally represented and in entorhinal cortex where 5-HT1E sites represented the major binding in layer III. They were also present in layers II and IV (29 and 24%) and, to a lesser extent, in layers V and VI. 5-HT1A sites were predominant in layer VI, II and V and were less abundant in other layers. 5-HT1D were homogeneously present in layers II, III, IV and were present in low amounts in other layers. No 5-HT1E were detected in choroid plexus, where [3H]5-HT was dramatically reduced by mesulergine (5-HT2C receptors). No significant displacement of [3H]5-HT by mesulergine was measured in other structures.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed1994

Autoradiographic localization of receptors for neuropeptide FF, FLFQPQRFamide, in human spinal sensory system.

The regional distribution of FLFQPQRFamide binding sites on fresh unfixed cryostate sections from post mortem specimens of human spinal cord and lower medulla oblongata was studied by quantitative autoradiographic methods using [125I]YLFQPQRFamide as ligand. Samples were taken from five cases who had died with no history of neurological disease at ages ranging from 5 months to 66 years. The biochemical and pharmacological characteristics of [125I]YLFQPQRFamide binding to mounted tissue sections were comparable to those reported for the rat in a previous study. [125I]YLFQPQRFamide appeared to interact reversibly with high affinity binding sites (Kd = 0.06 nM), distinct from opiate receptors. Sites labelled with [125I]YLFQPQRFamide were distributed unevenly within the human spinal cord and lower medulla oblongata, with the highest density in the superficial layers of the dorsal horn and the spinal trigeminal nucleus. Although moderate labelling was observed in the ventral part of spinal grey matter, dense labelling appeared in the gracile and cuneate nuclei. No binding sites were detected in white matter. These results show that, as in the rat, FLFQPQRFamide receptors in the human spinal cord and lower medulla oblongata, are mainly concentrated within spinal areas implicated in the analgesic action of opiates. The possible role of these receptors in modulating spinal nociceptive information is discussed with respect to the pharmacological effects of substances acting on FLFQPQRFamide receptors in animals.

Aged↗
PubMed1994

Localization and developmental pattern of vasoactive intestinal polypeptide binding sites in the human hypothalamus.

Using a quantitative in vitro autoradiographic approach, vasoactive intestinal polypeptide (VIP) binding site densities were compared in the post-mortem hypothalamus of human neonate/infant and adult. The densities were similar during development in most of the hypothalamic nuclei and areas examined underlying the stability of 125I-VIP binding sites in the post-mortem hypothalamus of young and adult individuals. However, the ventral part of the medial preoptic area, the medial, lateral, and supramammillary nuclei were characterized by an increase of 125I-VIP binding with age. In young and adult individuals, the highest densities of hypothalamic 125I-VIP binding sites were detected in the supraoptic and infundibular nuclei; the ependyma; the organum vasculosum of the lamina terminalis; the horizontal limb of the diagonal band of Broca; the ventral part of the medial preoptic area (in adult); the suprachiasmatic, paraventricular, and periventricular nuclei; and the medial and lateral mammillary nuclei in adult. Moderate densities were found in the vertical limb of the diagonal band of Broca, the bed nucleus of the stria terminalis, the ventral part of the medial preoptic area in neonate/infant, the medial and lateral mammillary nuclei in neonate/infant, the supramammillary nucleus in adult, the dorsal hypothalamic area, and the ventromedial nucleus. Low to moderate binding site densities were observed in the other hypothalamic regions of young or adult individuals. The nonspecific binding ranged from 15% of the total binding in the anterior hypothalamus to 20% in the mediobasal and posterior hypothalamic levels. Taken together, these results provide evidence for a large distribution of VIP binding sites in neonate/infant and adult human hypothalamus suggesting the implication of VIP in the development of this brain structure and the maintenance of its various functions.

Adult↗
PubMed1994

Encoding a post-operative coronary artery bypass surgery care plan in the Arden Syntax.

The Arden Syntax for medical logic modules (Arden) was used to test the feasibility of encoding large, complex care plans. The critical portions of an existing paper-based care plan for the management of patients following coronary artery bypass graft (CABG) surgery were encoded in Arden and an X-windows user-interface was developed. The Arden Syntax proved adequate for encoding all of the necessary functions of the care plan. The limitations of the current Arden Syntax and possible additions to Arden are discussed.

Artificial Intelligence↗
PubMed1994

Comparison of the effects of IVth ventricular administration of some tryptamine analogues with those of 8-OH-DPAT on autonomic outflow in the anaesthetized cat.

1 The present study compares the effects on representative autonomic outflows of IVth ventricular application of tryptamine analogues which act at 5-HT1 receptors with 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT). 2 Cumulative doses of 8-OH-DPAT, N,N-di-n-propyl-5-carboxamidotryptamine (DP-5-CT) and 5-carboxamidotryptamine (5-CT, 2.5-40 nmol kg-1), sumatriptan (10-160 nmol kg-1), indorenate (100-800 nmol kg-1), 5-hydroxytryptamine (5-HT, 20-640 nmol kg-1) both alone and in the presence of cinanserin (0.1 mg kg-1) were given into the IVth ventricle of cats which were anaesthetized with a mixture of alpha-chloralose and pentobarbitone sodium, neuromuscularly blocked and artificially ventilated. Recordings were made of arterial blood pressure, heart rate, renal, cardiac, splanchnic and phrenic nerve activities, femoral arterial flow, tracheal and intragastric pressures. 3 Central application of each of the agonists evoked significant falls in arterial blood pressure. In addition 8-OH-DPAT, DP-5-CT, 5-CT and 5-HT all evoked a differential inhibition of sympathetic nerve activities, with renal nerve activity being the most sensitive and cardiac nerve activity the least sensitive. In the dose-ranges used, administration of sumatriptan evoked reductions only in renal and splanchnic nerve activities whilst indorenate reduced activity in all three sympathetic nerves to a similar extent. 4. The effect of the agonists on heart rate was more inconsistent than the effects on sympathetic outflow.IVth ventricular application of 5-CT and sumatriptan were without effect on heart rate whilst 8-OH-DPAT, DP-5-CT, indorenate and 5-HT alone and in the presence of cinanserin all evoked significant bradycardias. However, whilst atropine partially reversed the bradycardias evoked by 8-OHDPAT and only slightly reversed those caused by indorenate, atropine was without effect on those evoked by DP-5-CT or 5-HT.5. None of the analogues tested had significant effects on gut motility, phrenic nerve discharge or tracheal pressure. 8-OH-DPAT, DP-5-CT, indorenate and 5-HT were without effect on femoral arterial conductance. However, following pretreatment with cinanserin, 5-HT evoked a significant reduction in femoral arterial conductance. At its highest dose, sumatriptan evoked a significant increase in femoral arterial conductance as did 5-CT at the 20 nmol kg-1 dose.6. It is concluded that the present data support the view that 5-HT1A receptors at the level of the brainstem are involved in the central sympathoinhibitory effects caused by intravenous administration of 5-HT1A agonists. Further, brainstem 5-HT1A receptors play an important role in the control of renal sympathetic outflow while brainstem 5-HT2 receptors are involved in the control of skeletal muscle and/or skin blood flow. Selective tryptamine agonists for 5-HT1A receptors differ from non-tryptamine agonists in that they do not cause an increase in central cardiac vagal tone.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed1994

Studies of neuroregulators in the brain stem of SIDS.

Some dysmaturity of neuroregulator neuronal systems may be responsible for brain stem disorders. These disorders may partly explain the mechanism of death in SIDS. The available data using microbiochemical assays, immunocytochemical techniques and autoradiographic methods seem to show anomalies of some monoaminergic and of some peptidergic systems, especially in the medulla oblongata. All these data need to be confirmed by further studies. It should be understood that one positive effect of such neuroanatomical study on SIDS is to gain 'normative' data on the human brain during development.

Brain Stem↗
PubMed1994

Scanning the structure and antigenicity of HPV-16 E6 and E7 oncoproteins using antipeptide antibodies.

The structure and antigenicity of the HPV-16 E6 and E7 oncoproteins was studied using a set of antisera against overlapping synthetic peptides. We report that antigenic, mobile regions of the native proteins, as defined by reactivity with antipeptide antisera, occur at the N-termini of both E6 and E7 proteins, corresponding to regions of known or suspected protein-protein interactions. The putative zinc finger domains were consistently non-reactive, despite computer predictions of relatively high antigenicity, suggesting that the proposed zinc finger regions are held in stable secondary structures that the peptides were not able to mimic. In E6, the linker region between the two zinc fingers was antigenic, indicating that the two zinc finger structures might be able to articulate relative to one another by a flexible linker region. The highly antigenic N-terminal region of HPV-16 E7 was also found to be antigenic in E7 of both HPV-11 and HPV-18, indicating that the E7 proteins of different HPV types have similar antigenic structures. The identification of antigenic regions of the E6 and E7 proteins should be therefore be useful in the design of site-directed antibodies against E6 and E7 for numerous HPV types.

Amino Acid Sequence↗
PubMed1993

Distribution of thyrotropin-releasing hormone binding sites: autoradiographic study in infant and adult human hippocampal formation.

The rostrocaudal distribution of thyrotropin-releasing hormone (TRH) binding sites was studied in the human hippocampus. Cryostat sections of the right and left hippocampi from 6 infants (2 h to 5 months of age) and 11 adults (24 to 92 years) were subjected to in vitro quantitative autoradiography using [3H]MeTRH as a ligand. A single class of high affinity [3H]MeTRH binding sites with an apparent dissociation constant in the nanomolar range has been shown both in the infant and the adult. The maximal number of these sites was higher in the infant. No significant difference was observed between the general patterns of the right and the left hippocampi when taking postmortem delay and age as parameters. The highest concentrations of [3H]MeTRH binding sites were localized in the uncinate gyrus, the uncal subiculum and in the whole length of the molecular layer of the dentate gyrus. The lowest densities were present in the ventral subiculum. The major difference observed between the infant and the adult appeared in the molecular layer of the dentate gyrus where the densities were two-fold higher in infants (189 +/- 6 versus 88 +/- 2 fmol/mg of tissue). The only marked difference in the distribution was localized in the caudal part of the body where no specific labeling was found in the presubiculum of the infant.

Adult↗
PubMed1993

Can the 5-HT2/1c agonist DOI cause differential sympatho-excitation in nerves supplying the heart in anaesthetized cats?

A comparison of the effects on sympathetic nerve activity to the heart of intravenous administration of the selective 5-HT2/1c agonist 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI) alone and in the presence of the peripherally acting 5-HT2/1c antagonist BW501C67 were made in alpha-chloralose anaesthetized cats. Activity in both cardiac sympathetic nerves running in the vagus and in both inferior cardiac nerves was simultaneously recorded. In addition renal and phrenic nerve activity, heart rate, arterial blood pressure, femoral arterial flow and tracheal pressure were also recorded. DOI evoked a rise in blood pressure and increased femoral arterial resistance in both groups of animals. In the BW501C67 pretreated animals, DOI had no effect on heart rate but caused a significant increase in all sympathetic nerve activities. In non-pretreated animals, however, the rise in blood pressure was associated with variable effects on sympathetic nerve activity, a significant rise only occurring in renal nerve activity. In these experiments DOI evoked a bradycardia. The variability in sympathetic nerve activity in the non-pretreated animals may have resulted from the rise in blood pressure evoking a baroreceptor-mediated central sympathoinhibition which would interfere with the central sympathoexcitatory effects of DOI. It is concluded that centrally, DOI will cause sympathoexcitation. In addition, intravenous DOI acting on 5-HT2 receptors on bronchial smooth muscle evokes bronchoconstriction as indicated by the very large rise in tracheal pressure in non-BW501C67-pretreated animals. If not controlled this severely compromises respiration leading to a large overestimate of the sympathoexcitatory effects of stimulation of central 5-HT2/1c receptors.

Amidines↗
PubMed1993

Absence of adrenergic neurons in nucleus tractus solitarius in sudden infant death syndrome.

Immunohistochemical study of catecholamine synthesizing enzymes tyrosine hydroxylase (TH) and phenylethanolamine-N-methyl transferase (PNMT) was performed in lower brain stem of 5 controls and 9 sudden infant death "syndrome" (SIDS) cases. No difference was noticed in TH immunoreactive neuronal groups. With anti-PNMT antibody, electively in nucleus gelatinosus (NG), a subnucleus of nucleus tractus solitarius, an absence of immunoreactivity was noticed. Catecholamine neuronal cell bodies in NG were present. The discussion favours a nonartefactual interpretation of data. A delay in maturation would be a possible explanation.

Brain Stem↗
PubMed1993

Short latency vestibular evoked potentials.

Auditory responses, including the well-characterized auditory brainstem response, have been used extensively in clinical investigations. Evoked responses have not been adequately developed to investigate the vestibular system. The purpose of this study is to describe a new method for the evaluation of short-latency vestibular evoked potentials in human subjects. Standard ABR equipment is used, with a customized solid-state modification of the triggering mechanism. Signal averaging is used to record responses to multiple linear decelerations. Results indicate the presence of a short-latency wave, which is absent in vestibular-deficient subjects. The literature is reviewed and illustrative cases are presented. We believe vestibular evoked potentials are a promising new modality in investigation of vestibular physiology.

Evoked Potentials, Auditory↗