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Biomedical subjects

A C Scott

Publications and source records attributed to A C Scott.

At least 37 records · Page 2Linked to original sources
PubMed1995

Western blot mapping of disease-specific amyloid in various animal species and humans with transmissible spongiform encephalopathies using a high-yield purification method.

SAF-protein, an amyloid, is the main constituent of scrapie-associated fibrils (SAF) and a specific marker for transmissible spongiform encephalopathies (TSE). Using an improved extraction method and Western blot detection, the disease-specific amyloid was found in various parts of the central nervous system of hamsters orally infected with scrapie, of squirrel monkeys orally infected with kuru, sporadic Creutzfeldt-Jakob disease (CJD) and scrapie, of human patients with sporadic CJD, of a sheep with natural scrapie and of a cow with bovine spongiform encephalopathy (BSE). In human CJD samples, the concentration of TSE-specific amyloid was estimated to be 1000- to 10 000-fold lower than in the central nervous system of hamsters with scrapie. The extraction method has a yield of 70% and allows Western blot detection of the TSE-specific amyloid in samples representing 1-10 micrograms of brain tissue from intracerebrally infected hamsters, as well as in individual spleens from hamsters with terminal scrapie infected by the intracerebral, oral or intraperitoneal route. A 20-100 mg sample of material is sufficient for the extraction of the pathological protein from different rodent, monkey, ovine, bovine and human tissues. The results reported here demonstrate the potential suitability of the method for the routine diagnosis of TSE as well as for the detailed analysis of distribution patterns of the TSE-specific amyloid in experimental approaches to the investigation of these diseases.

Amyloid↗
PubMed1994

Correlation between the results of a histopathological examination and the detection of abnormal brain fibrils in the diagnosis of bovine spongiform encephalopathy.

A statistical comparison was made between the results of the statutory neurohistopathological method for the post mortem diagnosis of bovine spongiform encephalopathy (BSE) and the detection of abnormal brain fibrils (SAF). A total of 773 suspect cases was examined by both methods; it comprised 531 animals born before the feeding of ruminant-derived protein to ruminant species was prohibited and 242 born after the ban. The relative sensitivities and specificities of the methods were calculated for the diagnosis of clinically suspected BSE. The agreement between the results of the methods was excellent for 331 of the cases born before the ban and for all the cases born after it. In these cases the samples were not autolysed. For the 200 cases in which autolysis was recorded there was poor agreement between the diagnostic methods and this was attributed to an apparently reduced specificity of the histopathological diagnosis in the autolysed material. Despite the potentially greater specificity of fibril detection in the diagnosis of scrapie-like disease, this study indicates that a reliance on fibril detection alone may result in some false negative diagnoses, probably owing to the inadequate sampling of the tissues.

Animals↗
PubMed1993

Scrapie associated fibril detection on decomposed and fixed ovine brain material.

Samples of cerebral cortex from eight scrapie affected sheep and two unaffected control sheep were stored for up to nine days at temperatures ranging from 18 degrees C to 29 degrees C. Scrapie associated fibrils (SAF) could be detected in proteinase K treated brain extracts from all the eight scrapie affected animals after five days storage and in six out of the eight after nine days storage. SAF could not be detected in any brain extracts from the two control animals. Formol saline fixed brain material from a further six scrapie affected and two clinically normal sheep, were also subjected to an extraction technique used to detect fibrils. No characteristic SAF were observed in any of these fixed samples. Long filamentous structures were observed in four of the fixed scrapie affected brain extracts and in one of the fixed unaffected control brain extracts.

Animals↗
PubMed1992

Feline spongiform encephalopathy: fibril and PrP studies.

The brains from 18 cats were examined for the presence of the fibrils and modified PrP protein which are molecular diagnostic markers for scrapie-like diseases. Thirteen cats were referred with clinical neurological signs potentially indicative of feline spongiform encephalopathy (FSE). Of these, five had histopathological changes of FSE, five had other lesions of the central nervous system, and in three the brain was normal. The remaining five cats had no clinical neurological signs and were selected as controls. Fibrils and modified PrP protein were found in the brains of the five cats with FSE and in one of the cats with neurological signs but no histopathological changes in the central nervous system. Fibrils were present in the absence of modified PrP in the brains of two cats, one with neurological signs and a histologically confirmed meningioma, and one with no neurological signs and a histologically normal brain.

Animals↗
PubMed1992

Scrapie-like encephalopathy in a greater kudu (Tragelaphus strepsiceros) which had not been fed ruminant-derived protein.

A 19-month-old greater kudu (Tragelaphus strepsiceros), whose dam had died 15 months earlier with spongiform encephalopathy, required euthanasia after developing severe ataxia and depression with an apparently sudden onset. No macroscopic abnormalities were detected on post mortem examination but a scrapie-like spongiform encephalomyelopathy was apparent on histopathological examination of brain and segments of spinal cord. Negative stain electron microscopy of proteinase K-treated detergent extracts of tissue from the brain stem revealed the presence of scrapie associated fibrils, and a 25 to 28 kDa band comparable with that identified as abnormal PrP (prion protein) from the brains of domestic cattle with spongiform encephalopathy was detected using rabbit antiserum raised against mouse PrP. The animal was born nine months after the statutory ban on the inclusion of ruminant-derived protein in ruminant feeds and, as no other possible sources of the disease were apparent, it appears likely that the infection was acquired from the dam.

Animal Feed↗
PubMed1992

Bovine spongiform encephalopathy: detection of fibrils in the central nervous system is not affected by autolysis.

The effect of autolysis on the electron microscopic detection of the characteristic abnormal fibrils, originally called 'scrapie-associated fibrils', was investigated in four different areas of the central nervous system (CNS) from 10 clinically suspect BSE cattle after post mortem delay and compared with the histopathological diagnosis. The tissues for fibril detection were subjected to controlled incubations to simulate autolysis. Fibril detection in all areas sampled from nine animals in which BSE was confirmed by histopathology was not affected by combined post mortem delays and specific controlled treatments. Detection of fibrils from the cervical spinal cord was no less sensitive than from brain areas. Fibrils were not detected in the one suspect case in which histopathology did not reveal lesions of BSE. The study confirms that fibril detection is of diagnostic value in BSE when post mortem autolysis renders CNS material unsuitable for histopathology.

Animals↗
PubMed1992

Isolation of a Lelystad virus-like agent from British pigs and scanning electron microscopy of infected macrophages.

Six, one-week-old gnotobiotic piglets were inoculated with tissues or sera collected from field cases of porcine reproductive and respiratory syndrome. The piglets showed little or no illness, and two that were necropsied at 8 and 9 days post infection appeared grossly normal. However, a Lelystad virus-like agent was isolated from most of the inoculated pigs using porcine alveolar macrophage cultures. Seroconversion to the Lelystad virus was observed and some animals developed microscopically detectable interstitial pneumonias. Scanning electron microscopy was used to study the in vitro cytopathic effect of the Lelystad virus on porcine alveolar macrophages.

Abortion, Veterinary↗
PubMed1992

An evaluation of the Thermafil endodontic obturation technique.

The purpose of this study was to evaluate the Thermafil endodontic obturation technique and to compare it with laterally condensed gutta-percha. Thirty-seven maxillary central incisors were similarly prepared and divided into groups. Seventeen were obturated with Thermafil and 17 with lateral condensation. Three served as controls. After vacuum staining, all teeth were cleared and apical dye penetration was evaluated by two independent observers. Average leakage values were 0.24 mm and 1.32 mm for Thermafil and 0.47 mm and 1.18 mm for lateral condensation. There was no significant difference between the techniques, although a difference between evaluators was noted. Final results point to the relative subjectivity of in vitro leakage studies. When comparing the obturation times of both techniques, the Thermafil technique averaged 2 min 56 s while lateral condensation took 3 min 26 s. Statistical analysis showed no significant difference in obturation times.

Analysis of Variance↗
PubMed1992

Membrane conductances involved in amplification of small signals by sodium channels in photoreceptors of drone honey bee.

1. Voltage signals of about 1 mV evoked in photoreceptors of the drone honey bee by shallow modulation of a background illumination of an intensity useful for behaviour are thought to be amplified by voltage-dependent Na+ channels. To elucidate the roles of the various membrane conductances in this amplification we have studied the effects of the Na+ channel blocker tetrodotoxin (TTX) and various putative K+ channel blockers on the membrane potential, Vm. 2. Superfusion of a slice of retina with 0.5-10 mM-4-aminopyridine (4-AP) depolarized the membrane and, in fifty of sixty-three cells induced repetitive action potentials. Ionophoretic injection of tetraethylammonium produced similar effects. 3. In order to measure the depolarization caused by 4-AP, action potentials were prevented by application of TTX: 4-AP was applied when the membrane was depolarized to different levels by light. 4-AP induced an additional depolarization at all membrane potentials tested (-64 to -27 mV). We conclude that there are 4-AP-sensitive K+ channels that are open at constant voltage over this range. 4. 4-AP slowed down the recovery phase of the action potential induced by a light flash by a factor that ranged from 0.51 to 0.16. This reduction could be accounted for by the reduction in a voltage-independent K+ conductance estimated from the steady-state depolarization. 5. After the voltage-gated Na+ channels had been blocked by TTX, exposure to 4-AP further changed the amplitude of the response to a small (approximately 10%) decremental light stimulus. The change was an increase when the background illumination brought Vm to potentials more negative than about -40 mV; it was a decrease when Vm > -40 mV. The data could be fitted by a circuit representation of the membrane with a light-activated conductance and a K+ conductance (EK = -66 mV) that was partly blocked by 4-AP. The voltage range studied was from -52 to -27 mV; neither conductance in the model was voltage dependent. 6. The responses to small changes in light intensity in the absence of TTX were mimicked by a model. We conclude that a voltage-dependent Na+ conductance described by the Hodgkin-Huxley equations can amplify small voltage changes in a cell membrane that is also capable of generating action potentials; the magnitude of the K+ conductance is critical for optimization of signals while avoiding membrane instability.

4-Aminopyridine↗
PubMed1991

Natural scrapie: detection of fibrils in extracts from the central nervous system of sheep.

Extracts from the cervical spinal cord and from the medulla, thalamus, cerebellum and cerebral cortex of the brains of 10 sheep, histopathologically confirmed as cases of scrapie, were examined by electron microscopy for the presence of scrapie-associated fibrils. Characteristic fibrils were observed in all the extracts except for that from the thalamus of one sheep. No fibrils were found in any extracts from three control sheep. A comparison of these results with a similar study of 22 cases of bovine spongiform encephalopathy (BSE) suggests that in cases of scrapie the area of the brain chosen for the detection of fibrils is less critical than in cases of BSE, in which fibrils are more readily extracted from areas of the brain stem.

Animals↗
PubMed1991

A comparison of morphine-perphenazine and midazolam on preoperative sedation and arterial oxygen saturation.

The effectiveness of midazolam and a mixture of morphine-perphenazine premedication to produce sedation and their effects on preoperative oxygen saturation (SaO2) were examined. Eighty-five patients whose SaO2 measured with a pulse oximeter was greater than 90% and who were not receiving narcotic sedatives or oxygen were randomized to three groups. Each patient had his SaO2 recorded before premedication with placebo (saline), midazolam 0.08 mg.kg-1 or morphine 0.15 mg.kg-1 with perphenazine 2.5-5.0 mg im. From 30-90 min later, prior to anaesthesia SaO2 was repeated, and a sedation score was obtained by a blinded observer using a seven point scale. Median sedation scores were greater for midazolam (4) than for morphine-perphenazine (2) and placebo (1) (P less than 0.0001). As well, there was a decrease in the SaO2 in the morphine-perphenazine group (1.7 +/- 2.7%, P less than 0.001) but not in the midazolam and placebo groups (0.1 +/- 2.3%, -0.8 +/- 2.1%). In conclusion midazolam produced greater sedation than morphine-perphenazine and placebo without effect on SaO2 whereas morphine-perphenazine showed a decrease in SaO2 preoperatively.

Adult↗