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Biomedical subjects

R A Hall

Publications and source records attributed to R A Hall.

At least 19 recordsLinked to original sources
PubMed1995

Glycosylation and antigenic variation among Kunjin virus isolates.

Previous studies have found Kunjin (KUN) virus isolates from within Australia to be genetically homogenous and that the envelope protein of the type strain (MRM61C) was unglycosylated and lacked a potential glycosylation site. We investigated the extent of antigenic variation between KUN virus isolates from Australia and Sarawak using an immunoperoxidase assay and a panel of six monoclonal antibodies. The glycosylation status of the E protein of each virus was also determined by N glycosidase F (PNGase F) digestion and limited sequence analysis. The results showed that KUN viruses isolated within Australia oscillated between three antigenic types defined by two epitopes whose expression was influenced by passage history and host cell type. In contrast an isolate from Sarawak formed a stable antigenic type that was not influenced by passage history and was distinct from all Australian isolates. PNGase F digestions of KUN isolates indicated that 19 of the 33 viruses possessed a glycosylated E protein. Nucleotide sequence of the 5' third of the E gene of selected KUN isolates revealed that a single base change in PNGase F sensitive strains changed the tripeptide N-Y-F (amino acids 154-156 of the published sequence) to the potential glycosylation site N-Y-S. Further analysis revealed that passage history also had a significant influence on glycosylation.

Amidohydrolases↗
PubMed1995

A novel complex formed between the flavivirus E and NS1 proteins: analysis of its structure and function.

We examined the structural features and functional significance of a novel complex which forms between the envelope (E) protein and nonstructural protein NS1 of Murray Valley encephalitis (MVE) virus. Western blot analysis of virus-infected C6/36 cell lysates revealed that the undenatured form of this E-NS1 complex was a heat-sensitive E-(NS1 dimer) complex. Furthermore, the E-NS1 complex was observed in cells infected with Kunjin, Japanese encephalitis, West-Nile and Kokobera viruses which indicates the complex is a common feature of flavivirus infection. E-NS1 complex which had been immunoaffinity purified from MVE-infected cell lysates or eluted from gel slices exhibited partial breakdown into the individual monomers, demonstrating that the complex arose from the association of E and NS1 proteins and was not a single polypeptide created from incomplete gene cleavage. Radioimmunoprecipitation and western blot analysis of MVE-infected cell lysates and culture fluid preparations collected at various times after infection revealed that the E-NS1 complex has a long half life, accumulates in the virus-infected cell with time and is not secreted into the extracellular fluid. We have postulated that the E-NS1 complex, or at least a major portion of the complex, is a non-specific aggregation with no functional significance in the viral life cycle.

Animals↗
PubMed1995

Thoracoscopic removal of mediastinal parathyroid adenoma.

Mediastinal parathyroid tissue hyperfunctions in as much as 25% of the patients with primary hyperparathyroidism, and this may be responsible for causing conventional operative procedures to fail in as much as one-third of the cases. When lesions prove to not be accessible through a cervical incision, or when a mediastinal adenoma is diagnosed before cervical procedures, median sternotomy and angiographic ablation have been considered the only options for removal. However, thoracoscopy has theoretic advantages over both. Two patients underwent successful thoracoscopic removal of a hyperfunctioning ectopic mediastinal parathyroid adenoma and their cases are presented here.

Adenoma↗
PubMed1995

Fungal pulmonary abscess in an adult secondary to hyperimmunoglobulin E (Job's) syndrome.

Job's syndrome is characterized by recurring bacterial infections of the skin and sinopulmonary tract. Laboratory evaluation reveals consistent elevation of circulating immunoglobulin E levels. The syndrome has been reported as a rare cause of bacterial pulmonary abscess and pneumatocele formation in childhood; here we present a case of cavitating fungal abscess in an adult with Job's syndrome.

Adult↗
PubMed1995

Retrograde cerebral and distal aortic perfusion during ascending and thoracoabdominal aortic operations.

BACKGROUND: Several alternative circulatory management techniques during thoracic aortic reconstruction have been implemented at this institution. This study was performed to assess whether retrograde cerebral perfusion during proximal aortic operations and distal aortic perfusion during thoracoabdominal aortic operations have improved outcomes. METHODS: A retrospective review of 156 patients undergoing elective and emergent operations of the thoracic aorta over the past 7 years was performed. Seventy-five patients underwent proximal aortic procedures: 22 with ascending aneurysms, 45 with type A dissections, and 8 with arch reconstructions. Eighty-one patients underwent descending thoracic or thoracoabdominal procedures: 26 with Crawford type I aneurysms, 18 with type II, 8 with type III, 8 with type IV, 11 with traumatic transections, and 10 with type B dissections. Outcomes measured were neurologic injury, renal failure, and mortality. RESULTS: For proximal aortic procedures, the stroke rate was 12% using cardiopulmonary bypass and 48% using hypothermic circulatory arrest. The addition of retrograde cerebral perfusion decreased the stroke rate to 0% (p < 0.01) and the mortality rate to 7.1% compared with 37% for hypothermic circulatory arrest (p < 0.05). For thoracic and thoracoabdominal aortic operations, straight cross-clamping resulted in a 27% rate of spinal cord injury and a 24% rate of renal failure, whereas the addition of distal aortic bypass resulted in a statistically significant reduction (p < 0.01) in neurologic injury to 7% and a notable, but not statistically significant, decrease in renal failure to 13%. Distal aortic bypass also reduced the mortality rate from 22% to 7% (p < 0.05). CONCLUSIONS: Retrograde cerebral perfusion decreases the stroke rate and mortality rate in proximal aortic operations and distal aortic perfusion decreases the rates of neurologic injury, renal failure, and mortality in thoracoabdominal aortic operations.

Aortic Dissection↗
PubMed1995

Immunodominant epitopes on the NS1 protein of MVE and KUN viruses serve as targets for a blocking ELISA to detect virus-specific antibodies in sentinel animal serum.

Two mosquito-borne flaviviruses, Murray Valley encephalitis (MVE) and Kunjin (KUN), are the aetiological agents of Australian encephalitis. MVE causes a severe and potentially fatal form of the disease while KUN is responsible for only a few relatively mild cases. Therefore it is important that serological tests used in flavivirus surveillance differentiate between infections with these two viruses. However, this has been hampered in the past by the close antigenic relationships between flaviviruses in traditional serological assays. An epitope blocking ELISA using MVE-specific and KUN-specific monoclonal antibodies (mAb) reacting to the non-structural protein NS1 of these viruses and a flavivirus group-specific mAb reacting to the envelope (E) protein was assessed for testing sentinel animals for seroconversion to specific flavivirus infections. Using these assays we were able to detect serum antibodies to a variety of flavivirus in laboratory infected rabbits, and naturally infected chickens and in the case of primary infections, differentiate those caused by KUN or MVE. These assays are now used routinely in our laboratory for testing chicken sera from sentinel flocks in the Kimberley and Pilbara regions of north Western Australia.

Animals↗
PubMed1994

Effects of cyclothiazide on synaptic responses in slices of adult and neonatal rat hippocampus.

The effects of cyclothiazide, a drug that blocks AMPA receptor desensitization, on synaptic responses were studied in field CA1 of hippocampal slices from adult and neonatal rats. Cyclothiazide (100 microM) reliably increased AMPA receptor-mediated field EPSP decay times in adult slices but only after prolonged (60-120 min) applications. In neonatal slices, 30 min applications of cyclothiazide were sufficient to produce large and long-lasting increases in response decay times. Since the effects of cyclothiazide on AMPA receptor binding properties were similar in adult and neonatal forebrain membranes, the data indicate that the slow action of the drug in adult slices is probably due to diffusion barriers. The prolongation of EPSP decay by cyclothiazide suggests that AMPA receptor-mediated synaptic responses in hippocampus are terminated by receptor desensitization.

Animals↗
PubMed1994

Kainate binding to the AMPA receptor in rat brain.

Displacement of [3H]AMPA and [3H]CNQX by kainate was measured in membranes and solubilized fractions from rat brain. In soluble fractions, plots of [3H]AMPA and [3H]CNQX binding displaced by kainate resulted in one-site fits with Ki values in the range of 1-3 microM. In membranes, plots of [3H]AMPA binding displaced by kainate resulted in graphs which were better fit by two-site regression analysis than by a one-site fit. The Ki value for the high-affinity component of these two-site fits was 3-9 microM and the low-affinity component Ki was in the range of 70-120 microM; similar values were determined for kainate displacement of [3H]CNQX. The presence of thiocyanate ions had no effect on kainate displacement of [3H]CNQX. Since the affinity for kainate of the presumed synaptic AMPA receptor is in the range of EC50 values for kainate determined from physiological studies, these data contribute further evidence for the idea that kainate binding to synaptic AMPA receptors may be responsible for many of kainate's physiological effects.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1994

Arboviruses causing human disease in the Australasian zoogeographic region.

Over 65 arboviruses have been reported from countries in the Australasian zoogeographic region, but only a few have been implicated in human disease. These include the flaviviruses Murray Valley encephalitis (MVE), Kunjin (KUN), Kokobera (KOK), and dengue, particularly types 1 and 2; the alphaviruses Ross River (RR), Barmah Forest (BF), and Sindbis (SIN); and the bunyaviruses, Gan Gan and Trubanaman. In this paper recent epidemiological and clinical results pertaining to these viruses are reviewed, with major emphasis on MVE and RR viruses. The extensive early studies of Australian arboviruses have been reviewed by Doherty [49, 50], and their ecology and vectors more recently by Kay and Standfast [87]. In addition, the biology of MVE and KUN [113] and RR [87, 114] viruses have been the subjects of more detailed reviews. The Australasian zoogeographic region is defined as countries east of the Wallace and Weber lines, two hypothetical lines in the Indo-Australian archipelago where the fauna of the Australasian and Oriental regions meet. Seroepidemiological studies of human arboviral infections have suggested that the Japanese encephalitis flavivirus and the chikungunya alphavirus occur only in the Oriental region, whereas the related MVE and RR viruses, respectively, are restricted to the Australasian region [85, 148]. Serological results from Wallacea, the zone between the Wallace and Weber lines, are not so clear-cut [85]. This review is therefore restricted to countries east of Wallacea, specifically New Guinea and Australia.

Animals↗
PubMed1994

AMPA receptor development in rat telencephalon: [3H]AMPA binding and western blot studies.

Telencephalic membranes from rats of different embryonic (E16, E19) and postnatal (P2, P7, P14, adult) ages were assessed for alpha-[3H]amino-3-hydroxy-5-methylisoxazole-4-propionic acid ([3H] AMPA) binding and for immunoreactivity levels of AMPA receptor subunits (GluR1, GluR2/3, and GluR4). In addition, the synaptic markers synaptophysin and NCAM140 (a neural cell adhesion molecule isoform) were examined by immunoblot. The density of [3H]AMPA binding sites increased steadily with advancing age. This increase was due mainly to the development of the large low-affinity component (KD = 400 nM) that dominates the [3H]AMPA binding profile of adult rat brain membranes. As resolved by two-site regression analysis, the high-affinity component (KD = 15 nM) of the [3H]AMPA binding increased by approximately twofold from E16 to adult, whereas the low-affinity component increased by 25-fold. Staining for GluR1 and GluR2/3 increased steadily with increasing age at all time points examined; synaptophysin and NCAM140 exhibited similar ontogenic immunostaining profiles. GluR4 immunoreactivity was first evident at P14 and increased by adulthood.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Cyclothiazide decreases [3H]AMPA binding to rat brain membranes: evidence that AMPA receptor desensitization increases agonist affinity.

The effects of cyclothiazide, a drug which blocks AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) receptor desensitization, were tested on binding of [3H]AMPA to rat brain membranes. Cyclothiazide reduced [3H]AMPA binding by lowering the apparent affinity of the AMPA receptor. The magnitude of the decrease was temperature dependent and greater for membrane-bound than for solubilized receptors. These data provide evidence that desensitization increases the affinity of the AMPA receptor for agonists and indicate that a significant percentage of AMPA receptors in conventional equilibrium binding assays are in a desensitized state.

Animals↗
PubMed1993

Single channel recordings of reconstituted AMPA receptors reveal low and high conductance states.

Glutamate receptors belonging to the AMPA (alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid) subclass were partially purified 30- to 60-fold from forebrain of adult rats and incorporated into planar bimolecular lipid membranes. The channel conductance associated with the reconstituted receptors was activated by kainate and AMPA in a manner that suggests cooperative binding of two to three agonist molecules is required to induce channel opening. This conductance was blocked by the specific antagonist DNQX (6,7-dinitroquinoxaline-2,3-dione). When the partially purified AMPA receptors were reconstituted by the tip-dipping method in asymmetric saline conditions ('outside-out configuration'), the addition of 300 nM AMPA to the pseudo-extracellular solution elicited single channel current fluctuations that were also inhibited by DNQX. Analyses of the currents revealed that the ion channels of reconstituted AMPA receptors have two distinct conductance levels of 12 and 60 pS with the great majority of receptors belonging to the former variety. These results suggest that reconstitution may be useful in identifying factors that regulate the binding and conductance properties of AMPA receptors.

Animals↗
PubMed1993

Diuretic and diet effect on Menière's disease evaluated by the 1985 Committee on Hearing and Equilibrium guidelines.

Fifty-four patients, diagnosed with Menière's disease and treated with diuretics and a low-salt diet, were evaluated retrospectively with the 1985 AAO/HNS Committee on Hearing and Equilibrium (CHE) guidelines for vertigo and hearing changes. The patient data base was also evaluated with other methods that helped determine the effectiveness of the 1985 AAO/HNS CHE guidelines. After 24 months of therapy, vertigo control was complete or substantial in 79% of the patients, limited or insignificant in 19%, and worse in 2% as evaluated by the CHE 1985 guidelines. Hearing improved in 35% of the patients, was unchanged in 29%, was worse in 22%, and could not be classified by CHE guidelines in 14%. Hearing was also evaluated by comparison of individual thresholds before medical therapy, and at 22 and 74 months after the start of medical therapy. We found a stabilization of low- and mid-threshold frequencies, with an average rate of hearing loss approximating 0 dB/yr with 74 months of followup. The results of this preliminary study suggest that diuretics and a low-salt diet may decrease the natural progression of sensorineural hearing loss in patients with Menière's disease. Compared with other methods of data analysis, the 1985 CHE guidelines lacked sensitivity to evaluate the hearing changes observed.

Academies and Institutes↗
PubMed1993

Thiocyanate equally increases affinity for two DL-alpha-amino-3-hydroxy-5-methylisoxazolepropionic acid (AMPA) receptor states.

The effects of potassium thiocyanate on the binding of DL-alpha-amino-3-hydroxy-5-methylisoxazolepropionic acid (AMPA), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), and L-glutamate to the AMPA receptor complex were studied in rat brain membranes. Thiocyanate increased the affinity of AMPA receptors for both AMPA and L-glutamate and decreased the affinity for CNQX. Displacement of [3H]CNQX binding by AMPA revealed that the affinity for AMPA of both the high and the low affinity states of the receptor increased in the presence of thiocyanate, whereas the proportion of the two states was unchanged. In soluble fractions, which exhibit an apparently homogeneous population of the high affinity state of the receptor, the affinity for AMPA was increased by a magnitude similar to that seen for the high affinity state in the membranes. These results indicate that thiocyanate enhances AMPA binding not by converting low affinity receptors into the high affinity state but by equally increasing the affinities of the high and the low affinity states of AMPA receptors.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1992

Mouse telencephalon exhibits an age-related decrease in glutamate (AMPA) receptors but no change in nerve terminal markers.

The central excitatory amino acid receptor selective for alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) was examined in brain tissue from mice at 3 and 25 months after birth. Antibodies against the rat GluR-A glutamate receptor subunit (selective for kainate and AMPA) labeled a mouse brain component of about M(r) 100,000. Telencephalic tissue from the older group of mice exhibited 31% less immunoreactivity towards this component as compared with that from the young group. Binding of [3H]AMPA also decreased with age in the telencephalon to an extent which was similar to the loss of receptor immunoreactivity. Scatchard analysis revealed that this reduction is due to a decrease in receptor density and not to a change in binding affinity. In contrast, there were only small age-related changes in AMPA receptor immunoreactivity and binding levels in the brain stem and cerebellum. Binding to dopamine, serotonin, or GABA receptors was not significantly reduced in the older mice. Since the nerve terminal markers synaptophysin and the SV2 glycoprotein were not detectably different in the two groups of mice, the age-related reduction in AMPA receptors is not likely to be due to a general decrease in synaptic density. These data suggest that glutamatergic neurotransmission mediated by AMPA-type receptors is selectively impaired with aging in the telencephalon.

Aging↗
PubMed1992

Evidence that high- and low-affinity DL-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) binding sites reflect membrane-dependent states of a single receptor.

Binding of DL-alpha-[3H]amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid ([3H]AMPA) to lysed rat brain membranes in the presence of potassium thiocyanate resulted in curvilinear Scatchard plots that could be resolved by regression analysis into a large low-affinity component and a small high-affinity component. Solubilization with Triton X-100 resulted in solubilized and nonsolubilized fractions that were considerably enriched in the high-affinity component and correspondingly reduced in the low-affinity component. It thus appears that solubilization converts low-affinity AMPA receptors into high-affinity receptors. Also, synaptic plasma membranes were found to be greatly enriched in the low-affinity form and deficient in the high-affinity form of the AMPA receptor. These experiments provide evidence for the hypothesis that the high- and low-affinity components of AMPA binding are interconvertible states of the same receptor rather than separate binding sites and that the conversion of these receptors from their native high-affinity state to the low-affinity state occurs on insertion of the receptors into synapses.

Animals↗
PubMed1992

Functional reconstitution of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors from rat brain.

Glutamate receptors belonging to the subclass specifically activated by alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) were solubilized from rat forebrain membranes with Triton X-100 and partially purified through a series of three chromatographic steps. Specific [3H]AMPA binding increased 30-60-fold during the isolation procedure. A protein band recognized by antibodies against specific amino acid sequences of the glutamate receptor-A subunit was enriched with each purification step; the molecular mass of this band (105 kDa) corresponded to that of cloned AMPA receptor subunits. Photoaffinity labeling of forebrain membranes with 6-cyano-7-[3H]nitroquinoxaline-2,3-dione, a specific antagonist of the AMPA receptor, labeled a single band that comigrated with the immunolabeled protein. On reconstitution of the partially purified material into bilayer patches, single-channel current fluctuations were elicited by 300 nM AMPA and blocked by 1 microM 6,7-dinitroquinoxaline-2,3-dione.

Animals↗