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A T Lee

Publications and source records attributed to A T Lee.

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

Three-point phase-contrast velocity measurements with increased velocity-to-noise ratio.

We describe a technique to increase the velocity-to-noise ratio (VNR) of phase-contrast magnetic-resonance velocity images based on making three measurements/flow encoding axis rather than the usual two. A phase-aliased high first moment data set and a nonaliased low first moment data set are acquired, and the high-moment data are phase unwrapped using the low-moment data. The VNR of the resulting measurement is given by that of the high-moment measurement and increases linearly with the first moment. A factor of 4 gain in VNR was observed with only a 50% increase in scan time. Thus, this method is a much more efficient way to increase VNR than simple averaging.

Cerebrospinal Fluid↗
PubMed1995

Motion artifacts in fMRI: comparison of 2DFT with PR and spiral scan methods.

Activation signals based on BOLD contrast changes consequent to neuronal stimulation typically produce cortical intensity differences of < 10% at 1.5T. Hemodynamically driven pulsation of the brain can cause highly pulsatile phase shifts, which in turn result in motion artifacts whose intensity is larger than the activation signals in 2DFT scan methods. This paper presents a theoretical and experimental comparison of the magnitude of such artifacts for 2DFT and two other methods using non-Cartesian k-space trajectories. It is shown that artifacts increase with TR for 2DFT methods, and that projection reconstruction (PR) and spiral methods have significantly reduced artifact intensities, because these trajectories collect low spatial frequencies with every view. The spiral technique is found to be superior in terms of efficiency and motion insensitivity.

Artifacts↗
PubMed1995

Discrimination of large venous vessels in time-course spiral blood-oxygen-level-dependent magnetic-resonance functional neuroimaging.

A technique is described for discriminating blood-oxygen-level-dependent (BOLD) signal changes originating from large venous vessels and those that arise from the cortical parenchyma based on examining the temporal delay of each pixel's response. Photic stimulation experiments were performed with a conventional 1.5 T scanner and correlated each pixel's time-course with sine and cosine functions at the frequency of the stimulus. It was found that the signal in pixels anatomically associated with gray matter was delayed between 4 and 8 s compared with the stimulus, whereas the signal in pixels correlated with visible vessels and sulci was generally delayed from 8 to 14 s. This larger delay is consistent with the longer time required for blood to reach the larger vessels. Finally, stimulus-induced NMR phase changes were observed for the largest vessels, which are attributed to bulk susceptibility shifts.

Brain↗
PubMed1995

A role for DNA mutations in diabetes-associated teratogenesis in transgenic embryos.

Congenital malformations are the leading cause of death in infants of insulin-dependent diabetic mothers. Although there are data to suggest that hyperglycemia itself is teratogenic, few mechanisms have been proposed to explain diabetic embryopathy. To address the possibility that DNA mutations play a role in the fetal malformations associated with diabetes, we developed a transgenic mouse model system to measure the mutation frequency of a neutral target gene, lacI, during embryonic development in a maternal hyperglycemic environment. Despite the short 21-day gestational period of the mouse, we observed a twofold increase in the mutant frequency of the lacI transgene in fetuses that developed in a mild diabetic environment (blood glucose > 8.3 mmol/l) compared with those that developed under normoglycemic conditions (blood glucose < 8.3 mmol/l). These data provide the first direct evidence of the genotoxic effect of diabetes in vivo and suggest a mechanism for the teratogenicity of the maternal diabetic environment.

Amino Acid Sequence↗
PubMed1994

Comparative analysis of DNA mutations in lacI transgenic mice with age.

Transgenic mice have been developed recently that contain copies of the well-defined mutagenesis reporter gene, lacI, in an integrated bacteriophage-based shuttle vector. The lacI gene, which is present in all cells of the mouse, can be excised specifically from isolated genomic DNA and efficiently packaged into bacteriophage particles after the addition of packaging extracts. Mutations of the lacI gene are easily detected by the derepression of beta-galactosidase resulting in blue plaques in the presence of X-gal. Originally developed as a short-term in vivo mutagenesis assay to screen genotoxic agents, we have used this system to measure naturally occurring DNA mutations as a function of chronological age. There was a linear increase in the presence of phenotypic lacI mutants from birth to 24 months (r = 0.731, P < 0.001), such that 24-month-old mice have accumulated approximately fourfold more mutants than newborn pups. Molecular analysis of these spontaneously arising DNA mutations showed them to result predominantly from base substitutions (80.6-98.5%) that were equally distributed between transitions and transversions. However, lacI mutations in animals > 3 months of age demonstrated a higher percentage of mutations (12-19.4% vs. 1.2%) resulting from discriminable size changes (> 20 bp) than was observed for mice 1-2 months old. Sequence analysis of mutations resulting from a > 20 bp size change revealed them to be due to a duplication of adjacent lacI sequence. These results indicate that there is a gradual accumulation of DNA mutations with age and that the types of mutations also are influenced by the age of the animal.

Aging↗
PubMed1993

Transposition of an Alu-containing element induced by DNA-advanced glycosylation endproducts.

Advanced glycosylation endproducts react with DNA and cause mutations and DNA transposition in bacteria. To investigate the mutagenic effect of advanced glycosylation in mammalian cells, plasmid DNA containing the lacI mutagenesis marker was modified by advanced glycosylation endproducts in vitro, transfected into murine lymphoid cells, recovered, and analyzed for mutations, plasmid size changes, and the presence of shared insertion sequences. An 853-bp host-derived DNA sequence, designated INS-1, was identified as an insertion element common to plasmids recovered from multiple independent transfections. Modification of DNA by advanced glycosylation increased by 60-fold the apparent frequency of INS-1 transposition: from 0.025% to 1.5%. The INS-1 element contains a 180-bp region that is homologous to the Alu repetitive sequence family. INS-1 was also observed to be present within larger insertional mutations and, in two cases, an apparently truncated version of INS-1 that lacks the Alu region was identified. These results demonstrate the experimental induction of DNA transposition involving mammalian chromosomal elements and suggest that advanced glycosylation may play a role in the formation of Alu-containing insertions that have been found to disrupt human genes.

Animals↗
PubMed1993

Paracrine control of gastric epithelial cell growth in culture by transforming growth factor-alpha.

Studying primary cultures of replicating canine oxyntic mucosal cells, we found evidence for modulation of cell growth by endogenous factors. [3H]thymidine incorporation into DNA was rapid with cells cultured in medium free of serum or added growth factors, and growth rates of these cultures were markedly dependent on plating density, indicating mitogenic control by soluble endogenous growth factors. Data indicated that endogenous transforming growth factor-alpha (TGF-alpha) exerted mitogenic control under the following conditions. 1) TGF-alpha was detected in the cultured cells by radioimmunoassay and immunohistochemistry. 2) TGF-alpha-like immunoreactivity and receptor reactivity were present in the medium in concentrations sufficient to exert mitogenic control. 3) Receptors for TGF-alpha and epidermal growth factor (EGF) were present in the cultures. 4) Immunoabsorption by a TGF-alpha-specific antisera reduced [3H]thymidine incorporation. TGF-alpha was localized to parietal cells by immunohistochemistry and cell separation. In contrast, combined [3H]thymidine autoradiography and immunohistochemistry with anti-TGF-alpha did not detect TGF-alpha in dividing cells. We conclude that parietal cell TGF-alpha exerts paracrine control of mucosal cell growth in vitro, and we speculate that this is an important paracrine mechanism in vivo.

Cell Count↗
PubMed1991

Induction of gamma delta transposition in response to elevated glucose-6-phosphate levels.

The nonenzymatic glycosylation of nucleic acids in vitro by the reducing sugars, glucose or glucose-6-phosphate, alters both physical and biological properties. Recent investigations have demonstrated that elevated intracellular levels of glucose-6-phosphate in glycolytic mutants of E. coli resulted in a concentration-associated increase in mutations of a target plasmid. The majority of the plasmid mutations were due to large (greater than 1 kb) insertions or deletions. We describe here the further analysis of mutant plasmids isolated from bacteria grown under conditions which were conducive to the intracellular accumulation of glucose-6-phosphate. We have found that a number of the insertional plasmid mutations were the result of the movement of the transposable element gamma delta from the host genome into the plasmid. The frequency of gamma delta transposition was also associated with the amount of glucose-6-phosphate accumulated in the bacterial cells. Furthermore, the presence of another transposable element, either Tn 5 or Tn 10 in the host genome increased the rate of gamma delta transposition without affecting its own movement. The observed increase in gamma delta transposition suggests a novel mechanism of induction by reducing sugars which may be the result of DNA modifications by reducing sugars.

Base Sequence↗
PubMed1991

Mitogenic response of canine fundic epithelial cells in short-term culture to transforming growth factor alpha and insulinlike growth factor I.

We report methods allowing the culture of rapidly dividing gastric epithelial cells to investigate the regulation of mucosal cell replication. Cells from canine fundic mucosa were dispersed by enzyme treatment, enriched by filtration and elutriation, and cultured on collagen gel in DMEM/F12 medium. After 48 h, greater than 95% of the cells displayed immunoreactivity with antibody to cytokeratin, an epithelial marker. The cells formed confluent monolayers by 72 h with a transmembrane resistance of 1,600 ohm.cm2 when mounted in a Ussing chamber indicating retention of epithelial cell characteristics. Calf serum (0.1-2%) produced a dose-dependent mitogenic effect evident by increases in [3H]-thymidine incorporation into acid-precipitated material and in cell number. After an 18-24-h incubation with [3H]-thymidine, approximately 55% of the cells cultured in 2% serum showed evidence of DNA synthesis by autoradiography and all of the replicating cells were cytokeratin positive. Using comparable culture conditions, a similar proportion of cells incubated for 18-24 h with bromodeoxyuridine displayed nuclear anti-bromodeoxyuridine immunoreactivity, thus indicating that over half of the cells in these cultures synthesized DNA during this period. As with serum, epidermal growth factor and transforming growth factor alpha (TGF alpha) (10 pM to 1 nM), insulin (10 nM to 1 microM) and insulinlike growth factor-I (IGF-I, 1-100 nM) increased [3H]-thymidine uptake. The greater potency of IGF-I, compared to insulin, suggests the presence of IGF-I receptors. We conclude that this culture preparation is composed of fundic mucosal epithelial cells and contains a predominance of dividing epithelial cells. EGF/TGF alpha and IGF-I are potential factors directly regulating proliferation of fundic mucosal cells.

Animals↗
PubMed1991

Aircrew decision-making behavior in hazardous weather avoidance.

In-flight encounters with hazardous weather represent one of the most significant safety issues in civil aviation operations. Aircrew judgment is often cited as the probable cause of incidents and accidents involving weather, although lack of information is also a factor. The present study examines how information, presented at different times and in different forms, affects the awareness and decision-making behavior of aircrews in a flight simulation study of a recent microburst/windshear incident. In order to examine the influence of enhanced information transfer on aircrew behavior, intracrew communications and approach-to-land decisions were evaluated with conventional ATC communications and with automated cockpit alerting and display of weather information. Results of the study revealed that aircrews provided only with conventional ATC transmissions of weather information had difficulty discriminating conditions conducive to microburst events from less hazardous windshear events. Improved situation awareness for microburst events was found when ground-based convective weather information was provided in real time to aircrews. Avoidance decision-making was found to be less efficient with conventional ATC alert transmissions when compared to the performance of crews provided with a visual display of microburst events. The importance of information transfer on aircrew situation awareness and decision-making in hazardous weather avoidance is discussed.

Adult↗
PubMed1990

In vitro and in vivo reactions of nucleic acids with reducing sugars.

Reducing sugars such as glucose and glucose 6-phosphate have been shown to nonenzymatically react with the amino groups of proteins. The modification of proteins by reducing sugars can alter both physical characteristics and biological functions. Analogous to the reaction observed with proteins, the amino groups of DNA bases are also able to react nonenzymatically with reducing sugars. The modifications of DNA by reducing sugars result in the time- and sugar-concentration-dependent changes in biological properties. In this communication we review data describing in vitro and in vivo models we have used to investigate the biological consequences of the nonenzymatic glycosylation of DNA.

Animals↗
PubMed1989

Antibodies to cachectin/tumor necrosis factor reduce interleukin 1 beta and interleukin 6 appearance during lethal bacteremia.

Cytokines secreted in response to invading micro-organisms are important mediators of detrimental hemodynamic and metabolic changes in the host. To test whether cachectin/TNF plays a role in triggering release of other cytokines in the setting of infection, anesthetized baboons were passively immunized against systemic cachectin/TNF before infusion of a LD100 dose of live Escherichia coli. Bacteremia led to significant increases in circulating levels of cachectin/TNF, IL-1 beta, and IL-6. Although bacterial endotoxin/lipopolysaccharide is a potent stimulus for the synthesis and release of IL-1 and IL-6 in vitro, specific neutralization of cachectin/TNF in vivo with mAb pretreatment significantly attenuated both the IL-1 beta and the IL-6 responses despite fulminant overwhelming bacteremia. These data suggest that cachectin/TNF is essential for the initiation or amplification of IL-1 and IL-6 release during lethal gram-negative septic shock syndrome.

Animals↗
PubMed1989

Interaction of the chicken progesterone receptor with heat shock protein (HSP) 90.

The chicken progesterone receptor A (PRA) was expressed from cDNA by in vitro transcription and translation and also by transient transfection of receptor-negative COS M6 cells. These receptors synthesized from cDNA exhibited functional properties similar to those of oviduct PRA. The ability of PRA to form an 8S complex and to bind to DNA was studied. PRA, synthesized by either expression system, formed an 8S complex which was dissociated by incubation in vitro with 0.4 M NaCl or 20 nM progesterone to generate a 4S species able to bind to DNA-cellulose. The presence of HSP 90 in the PRA 8S complex was confirmed by use of an HSP 90-specific antibody, AC-7. Expression constructs coding for various receptor deletions were studied in order to identify the site of interaction of PRA with HSP 90. Deletions of 290 amino acids from the C-terminus resulted in the loss of ability to form an 8S complex. Truncated receptor proteins lacking 153 amino acids from the C-terminus or 369 amino acids from the N-terminus were able to interact with HSP 90. These data suggest that the site of interaction between PRA and HSP 90 responsible for 8S complex formation may be in this region (amino acid 369-506). However, small internal amino acid deletions in this region of PRA did not result in the loss of interaction of mutant receptor proteins with HSP 90. Thus, it appears that there may be more than one site of interaction between PRA and HSP 90 in this region.

Animals↗
PubMed1989

Nonenzymatic glycosylation of DNA by reducing sugars.

The process of nonenzymatic glycosylation has proven to be not only of interest to food chemists in the preservation of foodstuffs, but has become of increasing interest to biologists. The nonenzymatic glycosylation of biologically relevant proteins has become an important factor in explaining some of the posttranslational modifications of proteins observed in diabetes and aging. Using in vitro and in vivo models, we describe in this paper the implications that the nonenzymatic glycosylation of DNA may have in the biological aging process.

DNA↗
PubMed1989

Flight simulator platform motion and air transport pilot training.

The influence of flight simulator platform motion on pilot training and performance was examined in two studies utilizing a B-727-200 aircraft simulator. The simulator, located at Ames Research Center, is certified by the FAA for upgrade and transition training in air carrier operations. Subjective ratings and objective performance of experienced B-727 pilots did not reveal any reliable effects of wide variations in platform motion design. Motion platform variations did, however, affect the acquisition of control skill by pilots with no prior heavy aircraft flying experience. The effect was limited to pitch attitude control inputs during the early phase of landing training. Implications for the definition of platform motion requirements in air transport pilot training are discussed.

Adult↗
PubMed1988

In vitro and in vivo studies of the effects of halogenated histidine analogs on Plasmodium falciparum.

The effects of four halogenated analogs of histidine on in vitro growth of Plasmodium falciparum malaria parasites were monitored by measurement of the incorporation of 3H-labeled amino acids into parasite proteins and by light and electron microscopy. The uptake of [3H]isoleucine was reduced to 50% of the control value by addition of 70 microM 2-fluoro-L-histidine (2-F-HIS) or 420 microM 2-iodo-L-histidine (2-I-HIS). [3H]histidine uptake into acid-insoluble material was affected equally by these two compounds, 50% inhibition resulting at 200 microM concentration. Morphological analysis of parasite development proved a sensitive assay, since development of mature trophozoites was inhibited 50% by 25 microM 2-F-HIS or 100 2-I-HIS. Electron microscopy studies suggested different mechanisms of action of 2-F-HIS and 2-I-HIS on P. falciparum. 2-F-HIS produced a decrease in knob number at the erythrocyte surface and accumulation of electron-dense material under the parasite membrane. 2-I-HIS had no obvious effect on knobs or electron-dense material but affected parasite morphology. Surprisingly, 2-chloro-L-histidine and 2-bromo-L-histidine did not inhibit P. falciparum in vitro, even though their halogen atom substituents are intermediate in size between F and I atoms. 2-F-HIS and 2-I-HIS were tested in vivo against P. falciparum in owl monkeys (Aotus sp.) but were ineffective at doses that were nontoxic.

Amino Acids↗