A role for lysosomes in scrapie pathogenesis.
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Biomedical subjects
Publications and source records attributed to J Lowe.
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The polymerase chain reaction (PCR) was used to identify spores of Bacillus anthracis. By using an assay capable of amplifying a 1247-bp fragment from the gene that encodes the edema factor of B. anthracis, as few as 10(3) copies of a plasmid containing the edema factor gene and as few as 2 x 10(4) spores were detected. Subjecting the product of this PCR to a second PCR designed to amplify a 208-bp fragment nested within the 1247-bp product improved detection to a single plasmid copy per PCR and to two spores of B. anthracis per PCR.
Forty-eight patients with retinitis pigmentosa completed at home a questionnaire about everyday tasks, and recorded their distance visual acuities on reduced Snellen charts. Respondents were asked to assess their own abilities for seven tasks involving visual search and mobility on a 1 to 4 scale. This revealed the relative difficulty of each task for the whole group. Other questions were open-ended to allow respondents to express their individual visual problems and experiences. Several problems came to light that were not amenable to treatment with low vision aids, but should be considered as a guide to the clinician in counseling.
Five patients with constricted visual fields and good central acuity were fitted with an experimental binocular field expander. They undertook a laboratory visual search task in a stationary horizontal 90 degree field with and without the device. Four of five patients adapted relatively quickly to the inherent optical distortions of the field expander. When this binocular aid was worn search efficiency improved significantly with practice; search time was highly correlated (r = -0.92; p less than 0.01) with information channel capacity in the larger visual field.
alpha B crystallin is a protein which has homology with the small cell stress proteins. A characterized antibody to residues 1-10 of alpha B crystallin was used to immunostain tissues containing ballooned (chromatolytic, achromasic) neurons. The tissues included two cases of classical Pick's disease, one case of dementia with swollen achromasic neurons in the cortex, two cases of Alzheimer's disease with large numbers of ballooned neurons, two cases of motor neuron disease, four cases of cortico-basal degeneration, and four cases with areas of brain showing swollen neurons adjacent to recent cerebral infarcts. The anti-alpha B crystallin showed strong diffuse cytoplasmic immunoreactivity of swollen cortical neurons in all the diseases. Astrocytes and oligodendroglial cells were also stained in normal tissues as previously described. Weak diffuse immunoreactivity with an antibody to ubiquitin-conjugates was also seen in the swollen neurons from cases of neurodegenerative disease but not following infarction. Ballooned neurons have been shown to contain phosphorylated neurofilament epitopes not normally present in the perikaryonal region. The presence of alpha B crystallin in ballooned neurons, together with previous data which also indicate its close association with intermediate filaments, suggest that alpha B crystallin may be involved in aggregation and remodelling of neurofilaments in disease. The presence of alpha B crystallin in neurons at the edge of areas of cerebral infarction is likely to reflect cells which are regenerating following damage; its detection may therefore be a marker for such cells. On a practical level, the antibody greatly facilitates the localization of such abnormal neurons in diagnostic histology.
A consecutive series is reported of 17 patients who underwent early surgical treatment for acetabular or unstable pelvic fractures associated with ipsilateral fractures of the femur. Treatment included external and internal fixation, and required careful consideration of the surgical approach and the positioning of the patient. The multiple injuries sustained by these patients required simultaneous procedures by several surgical teams. All the femoral fractures were internally fixed at the initial operation and eight patients had primary definitive treatment of all their other fractures as well. In nine patients the definitive treatment of their other fractures was delayed for an average of 11 days. There were no deaths, and no serious infections. The long-term morbidity resulted from the associated injuries and not from the pelvic or femoral fractures.
A retrospective study was carried out to identify factors which predisposed Thoroughbred horses to severe injuries, as compared to less severe injuries, while racing on New York Racing Association (NYRA) tracks during the period of January 1986 to June 1988. A severe injury was defined as an injury which led to humane destruction of the horse. A less severe injury was defined as a horse which didn't race within 6 months following a muscular, ligament, tendon, or skeletal injury on the racetrack. The data were obtained from the Horse Identification Department records kept by the Chief Examining Veterinarian of NYRA and included 55 severely injured horses and 245 less severely injured horses. Multiple logistic regression analysis was used to identify factors associated with the risk of severe injuries compared to less severe injuries in those horses. There was a significant association between track and the risk of severe injury (horses raced on Belmont and Saratoga were more likely to develop a severe injury compared to horses raced on Aqueduct Main). The track surface was also associated with the risk of severe injury (horses raced on a firm turf had a significantly lower risk of severe injury associated with the track was significantly modified by the track condition (horses raced at Belmont when it was muddy had a significantly increased risk compared to Aqueduct dirt). Horses were more likely to experience severe injury in the early part of the race (less than or equal to 6 furlongs) than the latter part of the race (greater than 6 furlongs). The risk of severe injury decreased with the age of the horse.
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Studies in recent years have shown that ubiquitin has increasingly important functions in eukaryotic cells; roles which were previously not suspected in healthy and diseased cells. The interplay between molecular pathological and molecular cell biological findings has indicated that ubiquitin may be pivotal in the cell stress response in chronic degenerative and viral diseases. Furthermore, the studies have led to the notion that ubiquitination may not only serve as a signal for nonlysosomal protein degradation but may be a unifying covalent protein modification for the major intracellular protein catabolic systems; these can act to identify proteins for cytosolic proteinases or direct intact and fragmented proteins into the lysosome system for breakdown to amino acids. This unifying role could explain why ubiquitin is restricted to eukaryotic cells, which possess extensive endomembrane systems in addition to a nuclear envelope. Protein ubiquitination is a feature of most filamentous inclusions and certain other intracellular conglomerates that are found in some degenerative and viral diseases. The detection of ubiquitin-protein conjugates is not of great diagnostic importance in these diseases. Protein ubiquitination is not only essential for the normal physiological turnover of proteins but appears to have been adapted as part of an intracellular surveillance system that can be activated by altered, damaged, or foreign proteins and organelles. The purpose of this system is to isolate and eliminate these noxious structures from the cell: as a cytoprotective mechanism this appears to have evolved in the cell akin perhaps to an 'intracellular immune system'. Other heat shock proteins such as hsp 70 may be involved in this process. It is apparent that ubiquitin has a role in embryonic development. Protein ubiquitination is presumably involved in the reorganisation of cytoplasm that accompanies cell differentiation. Ubiquitin is also necessary for the gross intracellular degradative processes which are consequent upon programmed cell death. Cell elimination is of key importance for a number of developmental morphogenetic changes. An understanding of the molecular details of these processes will no doubt provide further insights into the wide ranging roles of ubiquitin in the life process. As it says in the book 'Ubiquitin'; there is no doubt that ubiquitin is a 'lucky' protein. It is lucky in many ways: lucky for scientific progress, lucky for biomedical scientists and lucky for life! If you have not already done so, why don't you get lucky and look for a role for ubiquitin in your experimental system. As Avram Hershko has said "there is plenty to go round"!
Ubiquitin-protein conjugates are found in the primary (azurophilic) lysosome-related granules but not in the secondary (specific) granules in mature polymorphonuclear neutrophils prepared from bone marrow. This is the first reported demonstration of ubiquitin-protein conjugates in lysosome-related membrane-bound vesicles in granulocytes and complements our previous findings of ubiquitinated proteins in lysosomes of fibroblasts. The significance of the selective presence of conjugates in only one of the two main types of neutrophil granules remains to be elucidated but may relate to the presence of the complement of acid hydrolases, including proteases, in the azurophilic granules compared to the specific granules. Ubiquitin-protein conjugates may enter the primary granules during neutrophil maturation by an autophagic process or by a heterophagic process during the fusion of phagosomes with primary granules. Alternatively protein ubiquitination may be involved in granule biogenesis.
In 2 pairs of non-identical twins, haemorrhagic-shock encephalopathy syndrome developed in 1 co-twin while the other died of sudden infant death syndrome. The twin pairs were aged 3 and 4 months, respectively, and no cause was identified. We suggest that stress protein deficiency may underlie both syndromes.
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Immunofluorescence studies on Epstein-Barr virus (EBV)-transformed lymphoblastoid cells have previously shown that the latent membrane transforming protein (LMP-1) is found in patch-like inclusions which also immunostain for vimentin. We now show that EBV transformation causes a major reorganization of intermediate filaments, microtubules, mitochondria, and lysosomal elements, which generally become oriented around the microtubule organizing centre. Immunogold electron microscopy shows that LMP-1 is primarily concentrated in secondary lysosomes together with ubiquitin-protein conjugates and heat-shock protein 70. Intermediate filament inclusion formation with the above characteristics may be a general response triggered by other membrane glycoproteins; as seen, for example, in major human neurodegenerative diseases such as diffuse Lewy body disease.
Ten patients were diagnosed as having primary non-Hodgkin's lymphoma of the central nervous system at University Hospital, Nottingham, between September 1986 and April 1989. None had clinical evidence of HIV-1 infection. All the patients started treatment with chemotherapy (BVAM), designed to cross the blood-brain barrier, followed by radiotherapy. Seven patients completed both chemotherapy and radiotherapy. Dose reduction during chemotherapy was necessary in three patients because of neutropenia. In two of the six patients with solitary tumours, complete resection was achieved surgically prior to treatment. Five of the remaining eight patients (63%) had radiological evidence of a complete response with chemotherapy. The other three patients had no response to chemotherapy but one had a complete response after radiotherapy. The two-year cause-specific survival of the 10 patients was 37%. Two of the three patients who had a postoperative performance status of 0 or 1 (ECOG/WHO) are alive and disease-free at 26 and 46 months from diagnosis. The median survival of the seven patients with a performance status of 2-4 was 10 months with two patients alive and disease-free at 19 and 26 months. The two-year cause-specific survival of these seven patients was 22%.
An 82 year old patient was lost to follow-up having been diagnosed both clinically and on CT scan as having a right choroidal malignant melanoma. Twenty months later she presented with a painful right proptosis, dense cataract and raised intraocular pressure. A second CT scan showed orbital cellulitis with no evidence of the previous mass. Following enucleation histopathological examination revealed that the tumour had undergone total spontaneous necrosis. The implications of the CT scan findings are discussed. Five cases of necrosed choroidal malignant melanoma producing proptosis in the absence of extrascleral tumour extension have now been described.
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A case-control study was conducted on Thoroughbred horses to identify factors associated with the risk of breakdown on racetracks. A total of 310 cases (breakdowns) were identified from the Horse Identification Department records kept by the chief examining veterinarian of New York Racing Association. For each case, two control horses were selected randomly from the Daily Racing Form Inc. records. Multiple logistic regression analysis was used to identify and quantify the risk of factors associated with breakdown, while simultaneously controlling for the effect of other putative factors. Factors associated with risk of breakdown were: track (horses raced on Saratoga racetrack were at a lesser risk of breakdown), track composition/condition (turf tracks had a lower risk compared to dirt), number of seasons in race, racing in a later race, number of starts per year, the total number of starts, season (summer had a higher risk than winter or spring) and age of the horse.