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Biomedical subjects

D R Lee

Publications and source records attributed to D R Lee.

At least 55 records · Page 3Linked to original sources
PubMed1996

Initial biocompatibility studies of a novel degradable polymeric bone substitute that hardens in situ.

Clinical management of osseous defects often requires bone grafts. The standard for treatment is autogenous bone harvested from sites such as either the iliac crest or the outer table of the calvaria. In addition to the problem of donor site morbidity and the limited supply of graft material, there is the additional operating time associated with harvesting procedures. A synthetic, bone graft substitute that can match the clinical performance of autogenous bone could alleviate these deficiencies. Therefore, a polymeric bone substitute was developed that consists of a four-armed star polymer of poly(dioxanone-co-glycolide) endcapped at each termini with a biocompatible lysine-based diisocyanate crosslinker. The polymer can be mixed with inorganic fillers such as either hydroxyapatite or tricalcium phosphate to form either injectable or moldable putty. The addition of a catalyst (for example, diethylaminoethanol and water) to the polymer produces a crosslinking reaction causing the combination to harden. This reaction is nontoxic, normo-thermic and can be performed in situ. During the course of the polymerization, carbon dioxide is liberated, producing an interconnected porous network within the implant, suitable for bone ingrowth. This paper will describe a preliminary biocompatibility assay of the bone substitute.

Animals↗
PubMed1996

Identification of rhesus macaques with spontaneous endometriosis.

Rhesus macaques (Macaca mulatta) with endometriosis were identified using reproductive histories, serum levels CA-125, pelvic ultrasonography, laparoscopy, and histopathology. All animals were evaluated from a large breeding colony and had a history of infertility and/ or spontaneous abortions. Laparoscopy and ultrasonography were performed on 40 macaques: 27 macaques from the breeding colony with elevated CA-125 levels, ten macaques from the breeding colony with normal or low serum CA-125 levels, and three macaques from another facility with previously diagnosed spontaneous endometriosis. Clinical endometriosis was diagnosed by laparoscopy in 16/37 (43%) macaques from the breeding colony and was confirmed by histologic examination in all animals biopsied. The disease was classified as minimal (40%), mild (25%), moderate (10%), or severe (25%). The most common sites of endometriosis were the serosal surface of the uterus (75%) and the posterior cul-de-sac (75%). In this study, CA-125 levels were useful in identifying animals from the breeding colony with endometriosis. The rhesus macaque provides a valuable animal model to study endometriosis and potentially to assess efficacy of therapeutic agents for this disease condition.

Animals↗
PubMed1995

Peptide-induced rescue of serologic epitopes on class I MHC molecules.

To monitor conformational changes in MHC class I structure induced by interaction with peptide or beta 2-microglobulin (beta 2-m), we have taken a serologic approach. Previous studies by us and others have defined circumstances wherein specific peptides can decrease serologic recognition of class I molecules. However, such blocking of serologic epitopes has often been interpreted as steric hindrance by peptide side chains. In this paper, we describe peptide-induced gains in recognition by mAbs 30-5-7, 34-1-2, and B22/249. In experiments with mAb 30-5-7, impaired reactivity, which resulted from an Ld loop mutation, was specifically rescued by the binding of a beta-galactosidase-derived peptide to the Ld mutant. In studies with mAb 34-1-2, poor Ld detection was enhanced by mutations in Ld at beta 2-m interaction sites or by changes within the peptide-binding groove. To evaluate whether known peptides in the Ld groove could influence 34-1-2 recognition, we tested six peptide ligands, four of which increased the reactivity of 34-1-2 with the Ld-expressing cell to various degrees (up to 14-fold). It is of interest that Ld mutations at position 9 and 95/97 made significant differences in the ranking of the peptides in regard to their ability to increase recognition by 34-1-2 and B22/249. This finding suggests that mutations in the binding groove can alter peptide conformation and result in secondary changes in class I structure. On the basis of the cumulative serologic data, we propose that the class I molecule displays considerable fluidity, and is structurally influenced by both beta 2-m and peptide.

Amino Acid Sequence↗
PubMed1995

Disposition of misoprostol and its active metabolite in patients with normal and impaired renal function.

The disposition of misoprostol acid, the active metabolite of misoprostol, was studied in 48 subjects with various degrees of renal function after administration of a single 400 microgram oral dose of misoprostol. Subjects were assigned to one of four treatment groups: group 1, normal renal function with creatinine clearance (CLCR) 80-140 mL/min/1.73 m2; group 2, mild renal impairment with CLCR 50-79 mL/min/1.73 m2; group 3, moderate renal impairment with CLCR 20-49 mL/min/1.73 m2 or group 4, end stage renal disease (ESRD) patients maintained on hemodialysis. The maximum plasma concentration (Cmax) and time to reach Cmax (tmax) for misoprostol acid tended to be larger in group 4 subjects; however, it failed to reach statistical significance. Although not statistically significant, in group 4 subjects the terminal half-life (t1/2) of misoprostol acid was almost twice as large (1.27 +/- 0.77 h) as in groups 1, 2, and 3 (0.70 +/- 0.72, 0.72 +/- 0.67, and 0.73 +/- 0.45 h, respectively). Misoprostol acid's total area under the plasma concentration curve (AUC0 infinity) was larger in group 4 subjects (1173.5 +/- 487.4 pg.h/mL) as compared with groups 1, 2, and 3 (421.4 +/- 263.1, 418.9 +/- 114.5, and 377.0 +/- 145.2 pg.h/mL, respectively; P < .05). The apparent total body clearance (CL) of misoprostol acid was statistically significantly smaller in group 4 subjects (0.094 +/- 0.044 L/kg/min) as compared only with group 3 subjects (0.284 +/- 0.102 L/kg/min). The dose of misoprostol may need to be reduced in ESRD patients on prolonged hemodialysis to prevent unnecessary high plasma levels of misoprostol acid and to avoid possible dose-related adverse effects.

Acute Kidney Injury↗
PubMed1995

Twenty-four-hour ambulatory blood pressure monitoring in healthy elderly people: reference values.

One hundred and two healthy elderly volunteers (aged 65-83 years) underwent 24-hour ambulatory blood pressure monitoring. Mean day- and night-time pressures were higher than reported for younger subjects. Removing outlying BP measurements from individual 24-hour recordings only minimally affected mean figures of the group but in some individuals mean day diastolic BP may be up to 5 mmHg lower on editing. Using unedited data, mean day and night pressures were 134/81 mmHg and 119/67 mmHg respectively (corresponding 95th centiles 160/96 mmHg and 146/84 mmHg) and systolic and diastolic BP loads were 36% and 21%. These values may be used as temporary reference values for elderly subjects pending the results of longitudinal studies. Studies quoting ABPM data should specify whether data editing has been employed and, if so, the editing thresholds should be stated.

Aged↗
PubMed1995

The bioavailability and disposition of 1-(beta-D-arabinofuranosyl)-5-(1-propynyl)uracil (882C87), a potent, new anti-varicella zoster virus agent.

1. The bioavailability and disposition of 882C87, an anti-varicella zoster virus (VZV) agent, have been investigated in healthy young and elderly volunteers. 2. The mean bioavailability of a 200 mg tablet was 21.1% in the young (range 13.3-33.0%, n = 10) and 24.6% in the elderly (range 14.4-38.4%, n = 8), which is sufficient to achieve plasma concentrations well above the IC50 for anti-VZV activity. 3. Plasma concentrations of 882C87 after 50 mg i.v. were higher in the elderly than in the young, associated with a significantly longer half-life (13.7 vs 11.8 h) and decreased renal clearance (0.11 vs 0.14 ml min-1 kg-1) and total clearance (0.15 vs 0.17 ml min-1 kg-1). 4. After intravenous administration, the main route of elimination of 882C87 was renal with 81.6% recovered unchanged in urine in the young and 71.2% in the elderly. The pyrimidine base, 5-propynyluracil (5-PU) was unquantifiable in plasma and only present in trace amounts in urine. 5. After oral administration to four healthy volunteers, only 17% of a dose of [14C]-882C87 was recovered unchanged in urine and 58% as 5-PU, with total recovery in urine accounting for 86% of the dose. There was a lag of 4-12 h before the appearance of 5-PU in plasma, peak concentrations were one-third to a half those of 882C87. The data suggest that 5-PU is formed from unabsorbed 882C87 in the gut lumen and then absorbed and excreted in urine. 6. 882C87 is a potential once daily treatment for shingles.

Administration, Oral↗
PubMed1995

Investigation of ambulatory blood pressure monitoring data editing criteria.

Ambulatory blood pressure monitoring (ABPM) allows multiple measurements of BP over a 24h period but often produces a proportion of measurements which are distant from the mean and unlikely to be genuine. The software which accompanies the SpaceLabs 90207 monitor requires criteria to be set which excludes outliers (autoediting) but there is no published data evaluating at what levels these autoediting criteria should be set. This study set out to ascertain whether outlying BP measurements represent true blood pressures or machine error, the determinants of machine error and appropriate autoedit criteria. Twenty subjects, both normotensive and hypertensive, were studied by ABPM with a blinded observer recording BP using a standard mercury sphygmomanometer (SMS) connected via a T-tube during three activity phases: sitting with cuffed arm still, walking but keeping arm still during measurement and sitting but gently moving the arm during measurement. The results show that a high ABPM/SMS discrepancy is associated with arm movement. After studying different autoediting criteria which aimed to eliminate the maximum number of conflicting readings while excluding the minimum number of unconflicting readings, an autoediting threshold of between 1.75 and 2 s.d.s from the mean based separately on sleep and awake measurements is suggested.

Aged↗
PubMed1994

Association of Ld-restricted peptides with the wild-derived mouse Lw16 MHC class I molecule.

Previous serological analysis of untreated splenocytes and L cell transfectants expressing the wild-derived mouse major histocompatibility complex (MHC) class I molecule, Lw16, demonstrated the presence of two forms of this molecule in the cell lysates, one reactive with both the alpha 2 domain-reactive monoclonal antibody (mAb) 30-5-7 and the alpha 3 domain-reactive mAb 28-14-8 (30-5-7+ 28-14-8+), and the other reactive with only the latter of the two (30-5-7- 28-14-8+). Furthermore, the analysis suggested the presence of both forms on the cell surface. Due to the similarity of Lw16 to the inbred mouse-derived Ld molecule, we tested a panel of Ld-restricted and control peptides for their ability to bind to Lw16 molecules. Here, we report that two Ld-restricted viral peptides, lymphocytic choriomeningitis virus nucleoprotein (LCMV NP) 118-126 and murine cytomegalovirus (MCMV) pp89 168-176, significantly increase the number of Lw16 molecules on the cell surface as measured by the mAb 28-14-8, and the proportion of those molecules that are recognized by the mAb 30-5-7. This was further supported by an increase in mAb 30-5-7-reactive molecules in L.Lw16 cell lysates following treatment with either of these peptides. Examination of the stability of the different forms on the cell surface suggested that the 30-5-7+ Lw16 molecules induced with these peptides were unstable and probably lost their Ld-restricted peptides to generate 30-5-7- 28-14-8+ molecules; these latter molecules were also unstable. In contrast, putative 30-5-7+ and 30-5-7- 28-14-8+ Lw16 molecules on untreated cells were stable. Together, these results suggest that two Ld-restricted, viral peptides can induce assembly of or stabilize 30-5-7+ 28-14-8+ Lw16 molecules, mimicking endogenous self peptides. However, the association of the Ld-restricted peptides with Lw16 is apparently not optimal, since it results in unstable Lw16 molecules.

Amino Acid Sequence↗
PubMed1994

A polymorphic residue in the amino terminal alpha 1 hemi-domain of the mouse Ld class I molecule affects its assembly and surface expression.

In comparison to Dd and most other mouse major histocompatibility complex class I molecules, the Ld molecule is poorly expressed on the cell surface, has a lower affinity for beta 2-microglobulin and is trafficked more slowly to the cell surface. Previous studies using Ld-Dd exon-shuffled constructs and the chimeric Ddm1 molecules suggested that the Ld alpha 1 domain was responsible for this phenotype. Two constructs, one containing an Ld-Dd hemi-exon-shuffled alpha 1 exon and the other containing a Dd-Ld hemi-exon-shuffled alpha 1 exon, were inserted into either Ld or Dd to replace the intact alpha 1 exon. These constructs were transfected into mouse L cells. Flow cytometric analyses of the resulting transfectants indicate that the Dd-Ld alpha 1/Ld molecules, similar to the Dd alpha 1/Dd alpha 2/Ld molecules, were expressed at a higher level on the cell surface than either the Ld-Dd alpha 1/Ld molecules or intact Ld molecules. Analyses of the molecules in lysates suggested that a higher proportion of the Dd-Ld alpha 1/Ld molecules, like the Dd alpha 1/Dd alpha 2/Ld molecules, as compared to the Ld-Dd alpha 1/Ld and intact Ld molecules were assembled as detected by alpha 2 domain-reactive monoclonal antibodies. Pulse-chase and lysate stability studies suggested that the lower steady state levels of assembled Ld-Dd alpha 1 molecules resulted from a slower assembly rate rather than instability. Collectively, these studies suggest that residues in the amino terminal half of the Ld alpha 1 domain are responsible for its inefficient assembly, probably leading to its low cell surface expression. To determine which polymorphic residues in the amino terminal alpha 1 hemi-domain might influence this phenotype, several Ld point mutants, in which a Dd amino terminal alpha 1 hemi-domain residue was substituted into the corresponding position of Ld, were analysed. These analyses suggested that, while the residue at position 9 has only a slight effect on beta 2-microglobulin association, it has a striking effect on assembly and cell surface expression.

Animals↗
PubMed1994

Definition of TCR recognition sites on Ld-tum- complexes.

The P911 variant of the P815 mastocytoma was shown by Lurquin et al. (Cell 58:293, 1989) to elicit rapid tumor rejection in a syngeneic host. This rejection was mediated by Ld-restricted cytotoxic T lymphocytes (CTL) for which targets could be sensitized by the synthetic peptide designated tum- (P91A-.12-24). In a previous study, T cell clones specific for Ld-tum- complexes displayed very restricted TCR usage and a characteristic TCR motif in the V alpha CDR3 region, predicted to interact with peptide. However, in contrast to the majority of Ld peptide ligands that are nonamers, the tum- peptide is a 13-mer and its sequence does not fit the Ld binding motif. Thus, to define shorter versions of the tum- 13-mer and residues involved in TCR recognition, nonamer derivatives were synthesized and compared in several different binding and functional assays. From these comparisons, the peptide TQNHRALDL was found to be the optimal nonamer. CTL recognition of Ala-substituted analogues of this peptide indicated that the His and Arg residues at positions 4 and 5 are important for TCR contact. We propose that these basic residues of the tum- peptide interact with the previously defined acidic residues in the CDR3 region of several TCR known to recognize Ld-tum- complexes.

Amino Acid Sequence↗
PubMed1994

Amino acid composition of human milk is not unique.

To determine whether the amino acid pattern of human milk is unique, we compared the amino acid pattern of human milk with the amino acid patterns of the milks of great apes (chimpanzee and gorilla), lower primates (baboon and rhesus monkey) and nonprimates (cow, goat, sheep, llama, pig, horse, elephant, cat and rat). Amino acid pattern was defined as the relative proportion of each amino acid (protein-bound plus free) (in mg) to the total amino acids (in g). Total amino acid concentration was lower in primate milk than in nonprimate milk. There were commonalities in the overall amino acid pattern of the milks of all species sampled; the most abundant amino acids were glutamate (plus glutamine, 20%), proline (10%) and leucine (10%). Essential amino acids were 40%, branched-chain amino acids 20%, and sulfur amino acids 4% of the total amino acids. The amino acid pattern of human milk was more similar to those of great apes than to those of lower primates. For example, cystine was higher and methionine was lower in primate milks than in nonprimate milks, and in great ape and human milks than in lower primate milks. Because the milk amino acid patterns of the human and elephant, both slow-growing species, were dissimilar, the amino acid pattern of human milk seems unrelated to growth rate.

Amino Acids↗
PubMed1994

Economic comparison of nutritional management strategies for Venezuelan dual-purpose cattle systems.

Objectives of this study were to compare three nutritional management strategies for dual-purpose herds in Venezuela in 1987 using a deterministic, multiperiod linear programming model of a representative farm. The model maximized discounted net margin (total revenues minus variable costs) from the herd for a 3-yr cow replacement cycle partitioned into six periods. The periods accounted for seasonal variation in forage availability and quality, and the model provided information about optimal animal inventories, animal sales, land in forage, and feed supplements. We compared current nutritional management practices and alternatives allowing optimal supplementation with commercial concentrate, molasses, cassava root, and urea. Iteration between the programming model and results from the Cornell Net Carbohydrate and Protein System assured technical coefficients consistent with predicted animal performance. Compared with management practices relying on commercial concentrate, optimal use of molasses and urea permitted increases in the stocking rate. Productivity and profit were restricted primarily by energy intake, which was constrained by intakes of NDF and DM. Alternative management strategies changed the relative importance of nutrient requirements and feed intake constraints. Thus, optimal interventions to alleviate nutritional constraints will vary with current management. Mobilizing adipose and protein tissues during lactation was optimal for most strategies. Supplementing with molasses and urea instead of commercial concentrate was the most profitable strategy, increasing herd net margin by 16% compared to the predominant feeding strategy in the late 1980s.

Adipose Tissue↗
PubMed1993

Human T cell clones reactive against U-small nuclear ribonucleoprotein autoantigens from connective tissue disease patients and healthy individuals.

SLE and mixed connective tissue disease (MCTD) are characterized by the presence of high titers of autoantibodies against uridine-rich RNA-small nuclear ribonucleoprotein (snRNP) Ag. Because the presence of such snRNP-reactive autoantibodies has recently been shown to be associated with polymorphisms of HLA, this study was undertaken to determine whether snRNP-reactive T cells could be identified and characterized from patients. PBMC were stimulated with affinity-purified snRNP Ag and cloned by limiting dilution in the presence of rIL-2 and rIL-4, snRNP-reactive human T cell clones were generated from three patients and two healthy blood donors who possessed disease-associated HLA genotypes. The cell surface phenotype of clones determined by flow cytometry was CD3+, CD4+, CD45RO+, TCR V alpha beta+. TCR V beta analysis, performed using V beta-specific primers and polymerase chain reaction, revealed that the T cell lines generated were clonal; a limited number of TCR V beta genes were expressed among the clones tested. All clones tested by mAb blocking of Ag-induced proliferation were restricted by HLA-DR. Several T cell clones were identified that were specific for B'/B or D polypeptides. These results demonstrate that snRNP-reactive T cells can be isolated from SLE and MCTD patients in vitro, and that Ag-driven expansion of such T cells could play a role in the immunopathogenesis of these diseases in vivo.

Adult↗
PubMed1993

Binding of peptides lacking consensus anchor residue alters H-2Ld serologic recognition.

CTL recognize class I MHC/peptide complexes on the surface of target cells. Crystallographic and serologic data have indicated that peptide ligands can influence the conformation of class I molecules and hence T cell recognition. How the binding of peptides with disparate sequence motifs affects the conformation of distinct regions within a class I molecule remains unknown. A series of site-directed mutants of the murine class I molecule H-2Ld was studied to address this question. These mutants were generated by in vitro mutagenesis and used to map the serologic epitopes recognized by a panel of Ld-reactive mAb. The influence of six different ligands on serologic recognition by these mAb was then examined. Of 12 mAb tested, only one, B22/249, was found to be significantly influenced by the bound peptide. Peptide discrimination by B22/249 was observed at the cell surface and in immunoprecipitates of Ld after incubation with two of the six ligands. The two peptides that caused suboptimal B22/249 recognition of Ld/peptide lack a proline at position 2, which is present in the other four peptides and has previously been defined as an anchor residue for Ld ligands. The epitope on Ld detected by mAb B22/249 includes residues 63 to 70 on the alpha 1 domain helix. Two of these residues are in pocket B, which computer modeling predicts to be in contact with the second residue of Ld-binding peptides. Therefore, these data imply that a mAb to a class I molecule can distinguish peptides with different motifs, possibly reflecting peptide-dependent conformational changes in the class I molecule.

Amino Acid Sequence↗
PubMed1993

Slow egress of a mouse MHC class I molecule to the cell surface despite its strong association with beta 2-microglobulin.

Two H-2D region class I genes from the wild-derived mouse strain B10.GAA37 provisionally encoding the Dw16 and Lw16 molecules, respectively, were transfected into mouse L cells, and the expressed gene products were analyzed serologically by flow cytometry. As expected from nucleotide sequence comparisons, these analyses revealed that several Ld-reactive monoclonal antibodies (mAbs) recognize Lw16 and not Dw16. As detected by flow cytometry of intact L.Lw16 cells and B10.GAA37 splenocytes, and by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) of immunoprecipitates from splenocyte lysates, the alpha 2 domain-reactive mAb 30-5-7 detected less Lw16 than did the alpha 3 domain-reactive mAb 28-14-8, suggesting the existence of two populations of Lw16 molecules: 30-5-7+ 28-14-8+ and 30-5-7- 28-14-8+. Sequential immunoprecipitation studies provided further evidence for these two Lw16 subsets; furthermore, the 30-5-7- 28-14-8+ subset was found predominantly on the cell surface and in association with beta 2-microglobulin (beta 2-m). Pulse-chase studies of B10.GAA37 splenocytes revealed that Lw16, like Ld, is trafficked slowly to the cell surface, whereas Dw16 is trafficked quickly, like most other mouse K and D region class I molecules. Despite these similarities, Lw16 and Ld differ in their association with beta 2-m, in that the immunoprecipitates of Lw16 contained much higher levels of radiolabeled beta 2-m per heavy chain. Together, these studies indicate that the slower trafficking of Lw16 to the surface does not result from a weaker association with beta 2-m, suggesting that other factors, such as peptide ligand-induced assembly, and/or retention by ER-resident proteins play an important role in the trafficking of major histocompatibility (MHC) class I molecules to the cell surface.

Animals↗
PubMed1993

Clinic blood pressure measurements and blood pressure load in the diagnosis of hypertension.

We have retrospectively compared the blood pressure load derived from 24 hour ambulatory blood pressure monitoring in patients with all clinic blood pressure readings elevated with those with only some elevated pressures to establish whether clinic readings alone are good predictors of blood pressure status. Fifty-seven patients attending a district general hospital hypertension clinic who were not on anti-hypertensive treatment were selected. Between two and six clinic readings were taken over a period of 1-6 months. Forty out of 57 patients had at least one clinic diastolic blood pressure reading of < 90 mmHg and, of these, 14 (35%) had a high blood pressure load and 26 (65%) had a normal blood pressure load. Patients with all diastolic blood pressure readings > 90 mmHg totalled 17 and of these 11 (65%) had high load and six (35%) had normal load. Patients with clinic diastolic blood pressure > 90 mmHg were significantly more likely to be truly hypertensive on the basis of blood pressure load than if one or more clinic readings was below 90 mmHg (P < 0.05). Diastolic pressures have some predictive power as to the blood pressure status defined by blood pressure load, but even consistently raised diastolic pressures do not necessarily indicate hypertension. Likewise one or more clinic diastolic blood pressure < 90 mmHg does not assuredly indicate normotension. Twenty-four hour ambulatory blood pressure monitoring may have an increasingly important role in the assessment of hypertension.

Adolescent↗