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Biomedical subjects

J D Smith

Publications and source records attributed to J D Smith.

At least 145 records · Page 8Linked to original sources
PubMed1996

Tetrahymena thermophila: analysis of phospholipids and phosphonolipids by high-field 1H-NMR.

The phospholipids of control and lipid-modified Tetrahymena thermophila were identified and quantified, using 1-D and 2-D COSY proton NMR spectroscopy on intact lipids, before and after HPLC separation. The results are comparable to those obtained using classical lipid analytical techniques. The results indicate that the study of enzyme pathways and other metabolic processes involving phospholipids in Tetrahymena and related protozoa can be carried out using proton NMR spectroscopy as the investigating technique.

Animals↗
PubMed1996

Apolipoprotein E allele-specific antioxidant activity and effects on cytotoxicity by oxidative insults and beta-amyloid peptides.

The apolipoprotein E (APOE) E4 allele is associated with Alzheimer's disease, cardiovascular disease, and decreased longevity. To probe the mechanism of these associations, cell lines were created which secrete each apoE isoform. ApoE conditioned media, purified apoE, and commercially obtained apoE protected B12 cells from hydrogen peroxide cytotoxicity with E2 > E3 > E4. Physiological levels of apoE protected cells from beta-amyloid peptides, while higher doses of apoE led to increased cytotoxicity. E2 > E3 > E4 possessed antioxidant activity, and apoE bound certain metal ions. The decreased antioxidant activity of E4 could contribute to its association with Alzheimer's disease, cardiovascular disease and decreased longevity.

Alleles↗
PubMed1996

Alzheimer amyloid-beta peptide forms denaturant-resistant complex with type epsilon 3 but not type epsilon 4 isoform of native apolipoprotein E.

BACKGROUND: The apolipoprotein E (apoE) type epsilon 4 isoform specifies increased cerebral and cerebrovascular accumulation of amyloid-beta protein (A beta) and contributes to the genetic susceptibility underlying a large proportion (approximately 60%) of typical, sporadic Alzheimer disease. Unfortunately, in vitro biochemical studies of direct apoE isoform-specific interactions with A beta have been inconsistent, perhaps due to the use by different research groups of apoE isoform preparations in different conformational states (purified denatured versus native). MATERIALS AND METHODS: In the current study, we have investigated the possibility that synthetic A beta(1-40) preferentially associates with native apoE of either the type epsilon 3 or the type epsilon 4 isoform. RESULTS: Here, we demonstrate the preferential association of synthetic A beta(1-40) with native apoE epsilon 3. The complex between apoE epsilon 3 and A beta(1-40) could not be disrupted by sodium dodecyl sulfate. In a parallel assay, no denaturant-resistant association of A beta(1-40) with apoE epsilon 4 was detectable. CONCLUSIONS: These results support the notion that the apoE epsilon 4 isoform may foster beta-amyloidogenesis because apoE epsilon 4 is inefficient in forming complexes with A beta.

Amyloid beta-Peptides↗
PubMed1996

Novel neuritic clusters with accumulations of amyloid precursor protein and amyloid precursor-like protein 2 immunoreactivity in brain regions damaged by thiamine deficiency.

Experimental thiamine deficiency (TD) is a classical model of a nutritional deficit associated with a generalized impairment of oxidative metabolism and selective cell loss in the brain. In rats, TD-induced cell degeneration is accompanied by an accumulation of amyloid precursor protein (APP)/amyloid precursor-like protein 2 (APLP2) immunoreactivity in abnormal neurites and perikarya along the periphery of, or scattered within, the lesion. Prompted by these data and our previous findings of a genetic variation in the development of TD symptoms, we extended our studies to mice. C57BL/6, ApoE knockout, and APP YAC transgenic mice received thiamine-deficient diet and pyrithiamine injections. Unlike rats, APP/APLP2-immunoreactive neurites in all strains of mice were sparsely scattered within damaged areas and did not delimit the thalamic lesion. In addition, abnormal clusters of intensely immunoreactive neurites occurred only in areas of damage including the thalamus, mammillary body, and inferior colliculus. The clusters appeared as either irregular clumps or round or oval rosettes that strikingly resembled the neuritic component of Alzheimer amyloid plaques. However, immunostaining using various antisera to synthetic amyloid beta-protein (A beta 1-40) and thioflavine S histochemistry failed to show evidence of a component of A beta Neither APP/APLP2-immunoreactive clusters nor amyloid plaques were observed in the brain from patients with Wernicke-Korsakoff syndrome, the clinical manifestation of TD in man. Our results demonstrate species (i.e., genetic) differences in the response to TD-induced damage and support a role for APP and APLP2 in the response to brain injury. This is the first report that chronic oxidative deficits can lead to this novel pathology.

Amyloid beta-Protein Precursor↗
PubMed1996

Further characterization of the metabolic properties of triglyceride-rich lipoproteins from human and mouse apoC-III transgenic mice.

We previously showed that human apoC-III expression in transgenic mice causes hypertriglyceridemia due to the accumulation of enlarged very low density lipoprotein (VLDL)-like particles, with increased triglycerides and apoC-III and decreased apoE. In vivo turnover studies indicated the metabolic basis was decreased particle fractional catabolic rate. The presence of enlarged triglyceride-rich particles with prolonged residence time in plasma implied defective lipolysis, but in vitro these particles were good substrates for purified lipoprotein lipase (LPL). In the current study we further characterize the metabolic properties of these particles. We show that expression of a mouse apoC-III transgene can also cause hypertriglyceridemia with a similar accumulation of a VLDL-like particle with increased apoC-III and decreased apoE. A vitamin A fat tolerance test was used to show that MoCIIITg and HuCIIITg mice had similarly delayed clearance of triglyceride-rich postprandial particles. Thus, the previously observed hypertriglyceridemia caused by human apoC-III transgene expression was not due interspecies incompatibility but a property of apoC-III. In further experiments we showed VLDL from apoC-III transgenic mice interacted poorly with fibroblast lipoprotein receptors and this could be corrected by adding exogenous apoE. In addition, control VLDL interaction could be decreased by exogenous apoC-III. Moreover, the hypertriglyceridemia of HuCIIITg mice could be normalized by crossbreeding with HuETg mice. Thus, a functionally significant reciprocal relationship of apoC-III and apoE exists, presumably due to competition for space on the surface of triglyceride-rich lipoproteins. Finally, VLDL from HuCIITg and MoCIIITg mice showed decreased binding to heparin-Sepharose. This suggests and additional locus of the defect in these mice could potentially be in the binding of triglyceride-rich lipoproteins to heparan sulfate proteoglycan matrix on the surface of endothelial cells in which LPL is embedded. This could explain the predicted functional lipase deficiency in apoC-III transgenic mice based on the observation of a prolonged residence time of enlarged triglyceride-rich lipoproteins.

Animals↗
PubMed1996

Indicators of postoperative pain in cats and correlation with clinical criteria.

OBJECTIVE: To identify clinical indicators that may help identity postoperative pain in cats after ovariohysterectomy. ANIMALS: Healthy, laboratory animal source cats. PROCEDURE: Clinical indicators of pain were identified, and relief from pain in response to butorphanol was studied in 5 groups of cats. 10 cats had 1 hour of general anesthesia only, followed by recovery without additional medication. 10 cats had general anesthesia and ovariohysterectomy, followed by recovery without additional medication. 10 cats had general anesthesia, ovariohysterectomy, and postoperative administration of 0.1 mg of butorphanol/kg of body weight. Another 10 cats had general anesthesia, ovariohysterectomy, and postoperative administration of 0.3 mg butorphanol/kg. 10 cats received 0.1 mg of butorphanol/kg, IM, only. Samples and recorded data were obtained before, during, and after the anesthesia period. Clinical variables measured included heart rate, blood pressure, respiratory rate, rectal temperature, PCV, and blood glucose concentration. Results were compared with changes in norepinephrine, epinephrine, and cortisol concentrations. RESULTS: Cats that did not receive analgesics had higher cortisol concentration than did cats without surgery and cats that received butorphanol after surgery. Systolic blood pressure measured by ultrasonic Doppler was found to be predictive of cortisol concentration, using a multiple linear regression model. CONCLUSIONS: Cortisol concentration increased in response to surgical stress and pain, and this increase was diminished by use of butorphanol. CLINICAL RELEVANCE: Systolic blood pressure was the best clinical predictor of postoperative pain.

Analgesics, Opioid↗
PubMed1995

Cytochrome P450 1A1 promoter as a genetic switch for the regulatable and physiological expression of a plasma protein in transgenic mice.

Transgenic and gene knockout techniques allow for in vivo study of the consequences of adding or subtracting specific genes. However, in some instances, such as the study of lethal mutations or of the physiological consequences of changing gene expression, turning on and off an introduced gene at will would be advantageous. We have used cytochrome p450 1A1 promoter to drive expression of the human apolipoprotein E (apoE) gene in transgenic mice. In six independent lines, robust expression of the transgene depended upon injection of the inducer beta-naphthoflavone, whereas the seventh line had high basal expression that was augmented further by the inducer. The low level of basal expression in an inducer-dependent line was confirmed upon breeding the transgene onto the hypercholesterolemic apoE-deficient background. In the basal state transgene expression was physiologically insignificant, as these mice were as hypercholesterolemic as their nontransgenic apoE-deficient littermates. When injected with the inducer, plasma cholesterol levels of the transgenic mice decreased dramatically as apoE expression was induced to yield greater than physiological levels in plasma. The inducer could pass transplacentally from an injected mother to her fetuses with concomitant induction of fetal transgene mRNA. Inducer could also pass via breast milk from an injected mother to her suckling neonatal pups, giving rise to the induction of human apoE in neonate plasma. These finding suggest a strategy to temporarily ameliorate genetic deficiencies that would otherwise lead to fetal or neonatal lethality.

Animals↗
PubMed1995

Reduction of cellular rejection and increase in longer-term survival after heart transplantation after HLA-DR matching.

HLA matching in cardiac transplants is perceived as being logistically difficult. We studied 1135 consecutive primary cardiac allografts between 1980 and 1994 to assess the effect of HLA mismatching on long-term graft survival and cellular rejection episodes within 3 months of transplantation. We found a significant association between HLA-DR mismatching and the number of episodes of rejection (no mismatch 0.80 [SE 0.13], one mismatch 1.22 [0.06], two mismatches 1.42 [0.06], p < 0.05). We found a similar correlation between the total number of biopsy specimens showing evidence of cellular rejection and HLA-DR mismatch. The time between operation and the first rejection episode shortened with increasing HLA-DR mismatch (no mismatch 85.5 [37.3] days, one mismatch 43.1 [8.1], two mismatches 24.1 [2.9], p < 0.05). Furthermore, the proportion of patients with no evidence of rejection correlated with HLA-DR incompatibility. A significant association between improved graft survival and HLA-DR mismatching was found over 1, 5, and 10 years after transplantation (no mismatch 1 year 92%, 5 years 83%, 10 years 76%, one mismatch 1 year 81%, 5 years 73%, 10 years 59%, and two mismatches 78% 1 year, 5 years 70%, and 10 years 52%, p = 0.02). Increased efforts to prospectively HLA match patients has resulted in 25% of patients transplanted between January and May 1995 (n = 13/52) receiving grafts matched for HLA-DR. HLA matching reduces the frequency and severity of acute cardiac allograft rejection and improves graft survival for up to 10 years. Our preliminary results suggest that it is possible to use HLA matching prospectively for our selection of recipients.

Actuarial Analysis↗
PubMed1995

Decreased atherosclerosis in mice deficient in both macrophage colony-stimulating factor (op) and apolipoprotein E.

To develop a murine model system to test the role of monocyte-derived macrophage in atherosclerosis, the osteopetrotic (op) mutation in the macrophage colony-stimulating factor gene was bred onto the apolipoprotein E (apoE)-deficient background. The doubly mutant (op/apoE-deficient) mice fed a low-fat chow diet had significantly smaller proximal aortic lesions at an earlier stage of progression than their apoE-deficient control littermates. These lesions in the doubly mutant mice were composed of macrophage foam cells. The op/apoE-deficient mice also had decreased body weights, decreased blood monocyte differentials, and increased mean cholesterol levels of approximately 1300 mg/dl. Statistical analysis determined that atherosclerosis lesion area was significantly affected by the op genotype and gender. The confounding variables of body weight, plasma cholesterol, and monocyte differential, which were all affected by op genotype, had no significant additional effect on lesion area once they were adjusted for the effects of op genotype and gender. Unexpectedly, there was a significant inverse correlation between plasma cholesterol and lesion area, implying that each may be the result of a common effect of macrophage colony-stimulating factor levels. The data support the hypothesis that macrophage colony-stimulating factor and its effects on macrophage development and function play a key role in atherogenesis.

Animals↗
PubMed1995

Switches in expression of Plasmodium falciparum var genes correlate with changes in antigenic and cytoadherent phenotypes of infected erythrocytes.

Plasmodium falciparum expresses on the host erythrocyte surface clonally variant antigens and ligands that mediate adherence to endothelial receptors. Both are central to pathogenesis, since they allow chronicity of infection and lead to concentration of infected erythrocytes in cerebral vessels. Here we show that expression of variant antigenic determinants is correlated with expression of individual members of a large, multigene family named var. Each var gene contains copies of a motif that has been previously shown to bind diverse host receptors; expression of a specific var gene correlated with binding to ICAM-1. Thus, our findings are consistent with the involvement of var genes in antigenic variation and binding to endothelium.

Amino Acid Sequence↗
PubMed1995

Placement of a permanent cystostomy catheter to relieve urine outflow obstruction in dogs with transitional cell carcinoma.

Permanent cystostomy catheters were placed in 7 dogs to relieve urine outflow obstruction from presumed transitional cell carcinoma of the bladder trigone and urethra. The catheters were easily managed at home by the owners. Complications were minimal. The most frequent complication was urinary tract infection. Two owners complained of difficulty in draining the bladder 1 week before euthanasia of their dogs. The primary reason for euthanasia of all dogs was progression of the tumor. Survival times of these dogs were similar to those previously reported for dogs with transitional cell carcinoma involving the bladder and urethra, irrespective of treatment. Placement of a permanent cystostomy catheter should be considered in dogs with transitional cell carcinoma associated with urine outflow obstruction when, owing to the dog's condition or the owner's preference, radical surgery or other treatment is not an option.

Animals↗
PubMed1995

Humoral immune response to human aortic valve homografts.

The humoral response to homograft valves in humans is largely unknown. The anti-human lymphocyte antigen (HLA) antibody production, specificity, and immunoglobulin class were examined sequentially in 73 patients undergoing aortic valve replacement. In addition, the long-term production of antibodies was examined in a cross-sectional study of 160 patients at periods varying from 1 to 15 years postoperatively. Human lymphocyte antigen antibodies were produced in 17 of 30 antibiotic-sterilized homografts (56%) and in 15 of 15 "homovital" homograft recipients, compared with 6 of the 28 control xenograft recipients (21%) (p < 0.001). The HLA antibodies were immunoglobulin G in all 15 homovital homografts, in 11 of 17 antibiotic-sterilized homografts, and in four of the six xenograft cases. Human lymphocyte antigen specificities could be assigned to the antibodies in 21 cases. In 10 of 11 cases in which donor HLA typing data were available, the antibodies detected were directed against donor HLA class I antigens. Of six possible determinants of HLA antibody production, the type of homograft valve implanted (homovital or antibiotic sterilized) correlated with antibody formation. In the cross-sectional study, 66 of the 85 homovital homograft recipients tested for HLA antibodies after 1 year were found to have antibodies, compared with 29 of 75 antibiotic-sterilized homograft recipients (p = 0.00003). We conclude that homografts appear to stimulate a strong donor HLA-specific antibody response, particularly of the immunoglobulin G class. This is most common in homovital valve recipients. These antibodies can persist for 15 years after operation. The clinical significance of this response requires further investigation.

Antibody Specificity↗
PubMed1995

The role of subvocalization in auditory imagery.

Five experiments explored the utility of subvocal rehearsal, and of an inner-ear/inner-voice partnership, in tasks of auditory imagery. In three tasks (reinterpreting ambiguous auditory images, parsing meaningful letter strings, scanning familiar melodies) subjects relied on a partnership between the inner ear and inner voice, one similar to the phonological loop system described in the short-term memory literature. Apparently subjects subvocally rehearsed the imagery material, which placed the material in a phonological store that allowed the imagery judgement. In a fourth task (distinguishing voiced and unvoiced consonants in imagery), subjects still subvocally rehearsed, but seemed to need no additional phonological store to respond correctly. In this case they may have consulted articulatory or kinesthetic cues instead. In a fifth experiment (making homophone judgements), subjects hardly even needed to subvocally rehearse, a result suggesting that homophone judgements rely on some direct route from print to phonology. We consider the breadth of the partnership between the inner ear and inner voice, the level that subvocal rehearsal occupies in the cognitive system, and the functional neuroanatomy of the phonological loop system.

Acoustic Stimulation↗
PubMed1995

Characterization of class I MHC folding intermediates and their disparate interactions with peptide and beta 2-microglobulin.

Newly synthesized class I heavy chains achieve domain structure using disulfide bonds, assemble with beta-2 microglobulin (beta 2m), and bind peptide ligand to complete the trimeric complex. Although each of these initial events is thought to be critical for class I folding, their sequential order and effect on class I structure are unknown. Using mAb specific for distinct conformations of H-2Ld and Lq, we have defined folding intermediates of class I molecules. We show here that non-peptide-associated forms of Ld or Lq, detected by mAb 64-3-7 and designated L alt, lack numerous conformational epitopes surrounding their ligand binding sites. These results support the notion that L alt molecules have an open conformation. Interestingly, a significant proportion of L alt molecules were detected in association with beta 2m and these L alt/beta 2m heterodimers were preferentially folded by peptide in cell lysates. These findings indicate that class I heavy chain/beta 2m association can precede ligand binding and that peptide is probably the limiting factor for completion of the Ld/beta 2m/peptide trimeric complex in vivo. The characteristics of L alt molecules were investigated further by ascertaining the disulfide bond status of these molecules and their association with beta 2m and peptide. Treatment of cells with dithiothreitol (DTT), a membrane-permeable reducing agent, demonstrated that L alt molecules constitute a heterogeneous population including reduced, partially reduced and native class I molecules. Furthermore, partially reduced Ld alt molecules, in a cell line expressing a mutant Ld molecule lacking the alpha 2 domain disulfide bond, accumulated intracellularly, were not beta 2m-associated and displayed marginal peptide-induced folding in vitro. In accordance with this latter finding, peptide was found to preferentially convert fully disulfide-bonded forms of Ld alt to conformed Ld. Thus, we propose that intrachain disulfide bond formation precedes the association of class I heavy chain with beta 2m and peptide, and that disulfide bond formation is required for efficient assembly, ligand binding and folding of the class I heavy chain.

Animals↗
PubMed1995

Pre-transplant anti-epithelial cell antibodies and graft failure after single lung transplantation.

Eighty-five patients undergoing single lung transplantation have been studied to determine the presence of anti-epithelial cell antibodies (AECA) prior to transplantation using the human lung carcinoma epithelial cell line A549 in a microcytotoxicity assay. In addition, 29 healthy volunteers were also assayed for the presence of AECA. Twenty-seven of the 85 recipients exhibited AECA prior to transplantation compared to none of the 29 control subjects (p = 0.0001). Actuarial graft survival at 1 year was 78% for the AECA negative group compared to 56% for AECA positive recipients (p = 0.01). No correlation was seen between the presence of AECA and graft rejection as determined by transbronchial biopsy. However, there was an association between AECA and post-transplant infection (p = NS) where 16 (64%) of the AECA positive recipients had postoperative infection episodes compared to 25 (47%) of the negative recipients. Sodium dodecylsulphate polyacrylamide gel electrophoresis and Western blotting was also performed for 68 of the recipients and antibody reactivity was detected in 22 patients compared to 26 patients exhibiting AECA detectable by microcytotoxicity. The presence of AECA demonstrable by Western blotting did not correlate with graft survival, rejection or infection. In conclusion, AECA detectable prior to single lung transplantation are associated with a decrease in graft survival and with postoperative infections.

Adult↗
PubMed1995

The uncertain response in the bottlenosed dolphin (Tursiops truncatus).

Humans respond adaptively to uncertainty by escaping or seeking additional information. To foster a comparative study of uncertainty processes, we asked whether humans and a bottlenosed dolphin (Tursiops truncatus) would use similarly a psychophysical uncertain response. Human observers and the dolphin were given 2 primary discrimination responses and a way to escape chosen trials into easier ones. Humans escaped sparingly from the most difficult trials near threshold that left them demonstrably uncertain of the stimulus. The dolphin performed nearly identically. The behavior of both species is considered from the perspectives of signal detection theory and optimality theory, and its appropriate interpretation is discussed. Human and dolphin uncertain responses seem to be interesting cognitive analogs and may depend on cognitive or controlled decisional mechanisms. The capacity to monitor ongoing cognition, and use uncertainty appropriately, would be a valuable adaptation for animal minds. This recommends uncertainty processes as an important but neglected area for future comparative research.

Animals↗
PubMed1995

Computer screening for early detection of melanoma--is there a future?

Computer image analysis in the study of pigmented lesions is critically examined and discussed in the light of the current published data. The potential for objective analysis by computers as a possible screening aid for the inexperienced clinician is discussed. The future for this technology is exciting if handled with care.

Forecasting↗