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Biomedical subjects

J R Scott

Publications and source records attributed to J R Scott.

At least 19 recordsLinked to original sources
PubMed1992

Infection specific prion protein (PrP) accumulates on neuronal plasmalemma in scrapie infected mice.

Prion protein (PrP) is an abundant membrane-associated host protein which accumulates in abnormal, relatively protease-resistant forms in the brains of animals with scrapie and related diseases. Using correlative light and electron microscopy we determined the sites of subcellular localisation of PrP in mice infected with the 87V strain of scrapie. Disease specific accumulation of PrP was observed at light microscopy as amyloid plaques or as diffuse or granular staining within the neuropil, often clearly associated with individual neurons. Serial electron microscopical preparations were immunostained for PrP by the immunogold method. Gold particles were located on amyloid fibrils and on the plasmalemma of neurites at the periphery of plaques and in the neuropil, irrespective of the morphological form of PrP accumulation when viewed by light microscopy. This suggests that amyloid fibrils are formed following the accumulation and aggregation of sub-unit proteins at the plasmalemma and, furthermore, that normal PrP may be converted to its pathological form at this site.

Amyloid↗
PubMed1992

Ultrastructural features of spongiform encephalopathy transmitted to mice from three species of bovidae.

The ultrastructural neuropathology of mice experimentally inoculated with brain tissue of nyala (Tragelaphus angasi; subfamily Bovinae), or kudu (Tragelaphus strepsiceros; subfamily Bovinae) affected with spongiform encephalopathy was compared with that of mice inoculated with brain tissue from cows (Bos taurus; subfamily Bovinae) with bovine spongiform encephalopathy (BSE). As fresh brain tissue was not available for nyala or kudu, formalin-fixed tissues were used for transmission from these species. The effect of formalin fixation was compared with that of fresh brain in mice inoculated with fixed and unfixed brain tissue from cows with BSE. The nature and distribution of the pathological changes were similar irrespective of the source of inoculum or whether the inoculum was from fresh or previously fixed tissue. Vacuolation caused by loss of organelles and swelling was present in dendrites and axon terminals. Vacuoles were also seen as double-membrane-bound and single-membrane-bound structures within myelinated fibres, axon terminals and dendrites. Vacuoles are considered to have more than one morphogenesis but the structure of vacuoles in this study was nevertheless similar to previous descriptions of spongiform change in naturally occurring and experimental scrapie, Creutzfeldt-Jakob disease, Gerstmann-Sträussler-Scheinker syndrome and kuru. Other features of the ultrastructural pathology of the transmissible spongiform encephalopathies including dystrophic neurites and scrapie-associated particles or tubulovesicular bodies were also found in this study. Neuronal autophagy was a conspicuous finding. It is suggested that excess prion protein (PrP) accumulation, or accumulation of the scrapie-associated protease-resistant isoform of PrP, may lead to localised sequestration and phagocytosis of neuronal cytoplasm and ultimately to neuronal loss.

Animals↗
PubMed1992

Alloimmune conditions and pregnancy.

Transfer of fetal red blood cells and platelets to the maternal circulation can stimulate an immune response with production of immunoglobulin that can cross the placenta. Similarly, passage of maternal stem cells to an immunologically incompetent fetus can theoretically produce graft-versus-host disease. disease. Maternal sensitization to red blood cell antigens such as D and Kell can result in anaemia, hydrops, and death in an incompatible fetus. Current assessment of these pregnancies involves serial analysis of amniotic fluid bilirubin concentration, with umbilical cord blood sampling reserved for special circumstances; neither ultrasound or Doppler blood flow analysis are accurate in the prediction of fetal haematocrit. Intravascular transfusion is the treatment of choice for hydropic fetuses. Perinatal survival in non-hydropic fetuses is similar with either intravascular or intraperitoneal transfusion, and the choice of procedures is individualized. Isoimmune fetal thrombocytopenia is usually the result of maternal sensitization to the PlA1 antigen. There is significant risk of intracranial haemorrhage, both antepartum and during labour and delivery. Umbilical cord blood sampling at term can determine fetal platelet count and the need for platelet transfusion, and can aid in deciding the appropriate route of delivery.

Blood Transfusion, Intrauterine↗
PubMed1992

Anaerobic and aerobic power in arms and legs of elite senior wrestlers.

UNLABELLED: The purpose of this study was to characterize anaerobic power and aerobic power (power at peak oxygen uptake, or peak VO2) of elite senior (post-collegiate) wrestlers. Subjects (n = 14) from the U.S. national senior freestyle and Greco Roman wrestling teams were evaluated at separate tests using cycle ergometry for peak VO2 of the arms, peak VO2 of the legs, arm anaerobic peak power, and leg anaerobic peak power. Power output at peak VO2 was recorded for each test and compared to the anaerobic power of the appropriate body segment. The results (mean +/- SD) showed that wrestlers produced 2.3 +/- 0.4 W.kg body weight-1 at arm peak VO2 (43.7 +/- 4.6 ml.kg body weight-1. min-1), 4.2 +/- 0.5 W.kg body weight-1 at leg peak VO2 (50.9 +/- 5.1 ml.kg body weight-1.min-1), arm PP of 7.7 +/- 1.0 W.kg body weight-1, and leg PP of 10.5 +/- 1.7 W.kg body weight-1. Comparing power output during anaerobic and aerobic tests, the subjects performed at 3.4 +/- 0.6 times their peak oxygen uptake during arm anaerobic ergometry, whereas leg anaerobic peak power was 2.7 +/- 0.4 times the power at peak oxygen uptake for the legs (p < 0.05 for difference between ratios). CONCLUSION: relative to aerobic power, elite senior wrestlers may produce power anaerobically in the upper body at significantly higher levels than in the lower body.

Adult↗
PubMed1992

Scrapie in the central nervous system: neuroanatomical spread of infection and Sinc control of pathogenesis.

Following bilateral intraocular (i.o.) infection of Sinc s7 mice with ME7 scrapie, sequential tissue pools were taken from retina, optic nerve, superior colliculus (SC), dorsal lateral geniculate nucleus (dLGN), visual cortex and cerebellum. The infectivity levels in these pools were estimated by intracerebral (i.c.) assay in C57BL/FaBtDk mice. Infectivity was first detected in retina at 35 days post-injection (as an increase above residual injected inoculum), SC at 56 days, dLGN at 77 days and in optic nerve, visual cortex and cerebellum at 98 days. Pathological lesions were shown to develop in the same sequence later in the incubation period. Comparison of sequential retina and SC assays in congenic mice, which differ only in the vicinity of the Sinc locus, revealed a difference in the initial detection and progression of i.o. infection of between 60 and 100 days, indicating that Sinc acts by delaying the initiation of replication. Higher levels of infectivity were found in retina and SC of mice infected with 79A scrapie, which destroys the photoreceptor layer in the retina, than with ME7 scrapie, which does not. Retrograde transport of infection was indicated by the levels of infectivity in the retina after i.c. infection with ME7 or 79A scrapie. These results indicate that scrapie spread within the central nervous system is restricted to neuroanatomical pathways, and that Sinc controls the initiation, but not the rate of replication.

Animals↗
PubMed1992

CooB is required for assembly but not transport of CS1 pilin.

CS1 pili are filamentous proteinaceous appendages found on many enterotoxigenic Escherichia coli (ETEC) strains isolated from human diarrhoeal disease. They are thought to effect colonization of the upper intestine by facilitating binding to human ileal epithelial cells. We have identified a gene, cooB, which lies directly upstream of cooA, the gene that encodes the major structural CS1 protein. When translated in vitro, the protein product of cooB migrates in sodium dodecyl sulphate/polyacrylamide gel with an apparent molecular mass of 26 kDa, which is consistent with that predicted from its DNA sequence. We constructed a mutant allele (cooB-1) by insertion of the omega fragment, which inhibits transcription and translation, into the cooB gene in vitro. In a derivative of an ETEC strain with the cooB-1 mutation (JEF100) and a plasmid that encodes Rns (pEU2030), the positive regulator required for CS1 expression, no cooB and a greatly reduced level of cooA product was detectable in total cell extracts. The reduction of cooA in this strain appears to result from polarity of the cooB mutation because introduction of the wild-type cooA gene in trans causes production of CooA protein, which is found in cell pellet extracts, in extracts containing only surface proteins and in the culture supernatant. Therefore, in the absence of CooB, CooA is stable and it is transported through both inner and outer membranes. However, the cooB-1 strain with cooA in trans does not cause haemagglutination of bovine erythrocytes (the model system used to assay adherence mediated by coli surface antigen 1 (CS1) pili).(ABSTRACT TRUNCATED AT 250 WORDS)

Antigens, Bacterial↗
PubMed1992

Standardization of a technique for analysing the frequency of parasite-specific cytotoxic T lymphocyte precursors in cattle immunized with Theileria parva.

A limiting dilution microculture system was optimized to quantify the frequency of Theileria parva-specific cytotoxic T lymphocyte precursors (CTLp) in peripheral blood mononuclear cells (PBMC) from immune cattle. Optimal results were obtained with responder cell input levels ranging from 2 x 10(4)/well to 6.25 x 10(2)/well, along with 1-5 x 10(3)/well stimulator cells in standard supplemented RPMI 1640 medium containing 2.5-5% T cell growth factors. Thirty-six microtitre wells were established at each responder input level. Cultures were incubated for 7 days at 38 degrees C, at the end of which time individual wells were screened for cytotoxic activity in a 4-h 111indium oxine-release assay. Analysis of the cytotoxicity data, by a computer-programmed maximum likelihood estimation method indicated that they conformed to the Poisson model of single-hit kinetics. Estimates of frequencies ranged from 1:3600 to 1:5275 CTLp in PBMC of eight cattle between 1 and 24 months after immunization with T. parva. By contrast, no CTLp were detected in six naïve animals analysed to a responder cell input of 10(5)/well. Split-well analysis of individual microwells showed that the CTL clones generated under limiting dilution conditions displayed exquisite specificity for parasitized cells, were genetically restricted and in some animals were parasite strain-specific.

Animals↗
PubMed1992

Conjugative transposition of Tn916: the transposon int gene is required only in the donor.

Conjugative transposition of transposon Tn916 has been shown to proceed by excision of the transposon in the donor strain and insertion of this element in the recipient. This process requires the product of the transposon int gene. We report here the surprising finding that the int gene is required only in the donor during conjugative transposition. We find that Tn916 int-1, whose int gene has been inactivated by an insertion mutation, transposes when a complementing wild-type int gene is present only in the donor during mating. When the int+ gene is present in a plasmid and is expressed from the spac promoter, conjugative transposition is very inefficient. However, when the Int+ function is supplied from a coresident distantly linked Tn916 tra-641 mutant, which is defective in a function required for conjugation, efficient conjugative transposition of Tn916 int-1 occurs. This suggests either that Int is not required for integration of Tn916 in gram-positive bacteria or that the protein is transferred from the donor to the transconjugant during the mating event. When the nonconjugative plasmid pAT145 was present in the donor, it was rarely cotransferred with Tn916. This suggests that complete fusion of mating cells is not common during conjugative transposition.

Bacillus subtilis↗
PubMed1992

Environmental regulation of virulence in group A streptococci: transcription of the gene encoding M protein is stimulated by carbon dioxide.

We have found that different atmospheres can have significant effects on the transcription of emm, the gene that encodes M protein, the major virulence factor of the group A streptococcus (Streptococcus pyogenes). Expression of emm was monitored by constructing a transcriptional fusion of the promoter for emm6.1 from S. pyogenes JRS4 to a promoterless chloramphenicol acetyltransferase gene. Transcription, as measured by determining chloramphenicol acetyltransferase specific activity, was stimulated by as much as 25-fold by increased carbon dioxide tension. Expression was greater in the latter stages of growth and was not affected by growth at 30 instead of 37 degrees C. Insertional inactivation of mry, a gene encoding a positive regulator of emm6.1, reduced chloramphenicol acetyltransferase activity below the detectable level. We conclude that expression of emm is influenced by environmental factors and that the level of carbon dioxide is one signal that may influence expression of M protein during infection.

Antigens, Bacterial↗
PubMed1992

Transfer of Tn916 between Lactococcus lactis subsp. lactis strains is nontranspositional: evidence for a chromosomal fertility function in strain MG1363.

Lactococcus lactis subsp. lactis MG1363 can act as a conjugative donor of chromosomal markers. This requires a chromosomally located fertility function that we designate the lactococcal fertility factor (Laff). Using inter- and intrastrain crosses, we identified other L. lactis strains (LMO230 and MMS373) that appear to lack Laff. The selectable marker in our crosses was Tcr, carried by Tn916, a transposon present on the chromosome. The transfer of Tcr was not due to Tn916-encoded conjugative functions, because (i) L. lactis cannot act as a donor in Tn916-promoted conjugation (F. Bringel, G. L. Van Alstine, and J. R. Scott, Mol. Microbiol. 5:2983-2993, 1992) and (ii) transfer occurred when the Tcr marker was present in a Tn916 derivative containing a mutation, tra-641, that prevents Tn916-directed conjugation in any host. In addition, we isolated a strain in which Tn916 appears to be linked to Laff; this strain should be useful for further analysis of this fertility factor. In this strain, Tn916 is on the same 600-kb SmaI fragment as Clu, a fertility factor previously shown to promote lactose plasmid transfer in L. lactis. Thus, it is possible that Clu and Laff are identical.

Base Sequence↗
PubMed1992

Outcome of children born to women treated during pregnancy for the antiphospholipid syndrome.

OBJECTIVE: To determine the rate of neonatal and childhood medical complications in the offspring of women with the antiphospholipid syndrome who are treated during pregnancy. METHODS: We compared 29 infants born to 23 mothers with antiphospholipid syndrome with a group of control children matched for year and gestational age at birth and route of delivery. During pregnancy, mothers with antiphospholipid syndrome were treated with accepted therapeutic regimens, including prednisone and low-dose aspirin, heparin and low-dose aspirin, and others. RESULTS: Neonatal complications in the study infants included hyperbilirubinemia (14), respiratory distress syndrome (three), bronchopulmonary dysplasia (two), necrotizing enterocolitis (two), intraventricular hemorrhage (two), neonatal sepsis (one), coarctation of the aorta (one), hypothyroidism (one), hypoglycemia (one), and death (one). Two children had feeding problems, four had delayed psychomotor development, and eight were small for their age. However, the overall rate of neonatal or childhood complications did not differ from that in the control children. CONCLUSIONS: Our data indicate that prematurity and neonatal problems are common, but the childhood course for these offspring is similar to that of other premature infants.

Antiphospholipid Syndrome↗
PubMed1992

Outcome of treated pregnancies in women with antiphospholipid syndrome: an update of the Utah experience.

OBJECTIVE: To determine the outcome of treated pregnancies in women with well-characterized antiphospholipid syndrome. METHODS: We reviewed 82 consecutive pregnancies in 54 women with antiphospholipid syndrome who were treated during pregnancy with the following: 1) prednisone and low-dose aspirin; 2) heparin and low-dose aspirin; 3) prednisone, heparin, and low-dose aspirin; or 4) other combinations of these medications or immunoglobulin. RESULTS: The overall neonatal survival rate was 73%, excluding spontaneous abortions, but treatment failures (fetal and neonatal deaths) occurred in all treatment groups. Patients with successful treated pregnancies had fewer previous fetal deaths than those with unsuccessful treated pregnancies. There were no significant differences in outcome among the four treatment groups. Preeclampsia and fetal distress occurred in half of all pregnancies, and fetal growth impairment occurred in nearly one-third. Preterm delivery due to maternal or fetal indications was required in 37% of the pregnancies. Four pregnancies were also complicated by postpartum thrombosis during treatment. CONCLUSIONS: Pregnancy in women with antiphospholipid syndrome appears to be improved by treatment, but fetal loss may occur despite treatment. Preeclampsia, fetal distress, fetal growth impairment, and premature delivery are common. Because of the clinically significant risk of thrombotic episodes, thrombosis prophylaxis should be considered in these patients.

Antiphospholipid Syndrome↗
PubMed1992

Lateral cervical radiographs and adenoid size: do they correlate?

Clinicians have questioned the value of lateral soft tissue neck x-ray (LSTN) in assessing adenoid size. Elaborate cephalometric assays have been devised to measure degree of nasopharyngeal obstruction secondary to adenoid hypertrophy. This study prospectively studied 73 children, aged 11 months to 13 years, with clinical evidence of adenoid hypertrophy to assess how well a LSTN correlates with direct intraoperative observation of adenoid size and nasopharyngeal obstruction. We found a relatively weak correlation (Pearson coefficient r = 0.34) between x-ray and operative observations. We conclude that LSTN is an appropriate examination in the preoperative assessment of children being considered for adenoidectomy. However, this test must be interpreted by recognizing the inherent limitations of evaluating a dynamic structure, such as the nasopharynx, through a non-dynamic modality.

Adenoids↗
PubMed1991

Evidence for intrinsic control of scrapie pathogenesis in the murine visual system.

Using the optic nerve to route scrapie infection into the brain reduces the initial spread of the disease to well-defined neuronal relays, and simplifies the observation of cause and effect of agent transport, replication and degeneration of the nervous system. One drawback of intraocular targeting of infection is the relatively long incubation periods required to produce clinical disease. By using highly-enriched fractions of infectivity and two models of murine scrapie, we have found that this time delay is not simply due to the limited amount of infective inoculum that can be injected into the eye. This provides evidence of intrinsic control of scrapie pathogenesis within the murine visual system.

Animals↗