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Biomedical subjects

M Lee

Publications and source records attributed to M Lee.

At least 505 records · Page 28Linked to original sources
PubMed1996

Cannabinoid inhibition of adenylate cyclase-mediated signal transduction and interleukin 2 (IL-2) expression in the murine T-cell line, EL4.IL-2.

Cannabinoid receptors negatively regulate adenylate cyclase through a pertussis toxin-sensitive GTP-binding protein. In the present studies, signaling via the adenylate cyclase/cAMP pathway was investigated in the murine thymoma-derived T-cell line, EL4.IL-2. Northern analysis of EL4.IL-2 cells identified the presence of 4-kilobase CB2 but not CB1 receptor-subtype mRNA transcripts. Southern analysis of genomic DNA digests for the CB2 receptor demonstrated identical banding patterns for EL4.IL-2 cells and mouse-derived DNA, both of which were dissimilar to DNA isolated from rat. Treatment of EL4.IL-2 cells with either cannabinol or Delta9-THC disrupted the adenylate cyclase signaling cascade by inhibiting forskolin-stimulated cAMP accumulation which consequently led to a decrease in protein kinase A activity and the binding of transcription factors to a CRE consensus sequence. Likewise, an inhibition of phorbol 12-myristate 13-acetate (PMA)/ionomycin-induced interleukin 2 (IL-2) protein secretion, which correlated to decreased IL-2 gene transcription, was induced by both cannabinol and Delta9-THC. Further, cannabinoid treatment also decreased PMA/ionomycin-induced nuclear factor binding to the AP-1 proximal site of the IL-2 promoter. Conversely, forskolin enhanced PMA/ionomycin-induced AP-1 binding. These findings suggest that inhibition of signal transduction via the adenylate cyclase/cAMP pathway induces T-cell dysfunction which leads to a diminution in IL-2 gene transcription.

Adenylyl Cyclase Inhibitors↗
PubMed1996

Suppression of the hypersensitive response in potato by acetyl salicylic acid.

Increased salicylic acid has been correlated with systemic acquired resistance in several plants. Inoculation of potato plants with the pathogen Phytophthora Infestans or inducers from the fungus, arachidonic acid and eicosapentaenoic acid also induce systemic acquired resistance in the plant. We now report that treatment of potato tuber slices with acetyl salicylic acid markedly reduces the resistant response of these tissues.

Antifungal Agents↗
PubMed1996

Chymase in exocytosed rat mast cell granules effectively proteolyzes apolipoprotein AI-containing lipoproteins, so reducing the cholesterol efflux-inducing ability of serum and aortic intimal fluid.

Degranulated mast cells are present in human fatty streaks. Chymase in granules released from degranulated rat serosal mast cells, i.e., in granule remnants, proteolyzes human high density lipoprotein3 (HDL3), and so reduces its ability to induce cholesterol efflux from macrophage foam cells in vitro. In this study we found that remnant chymase, by proteolyzing human serum and human aortic intimal fluid, prevents these two physiologic fluids from effectively inducing cholesterol efflux from cultured macrophage foam cells. Inhibition was strongest when remnants were added to apolipoprotein AI (apoAI)-containing lipoproteins; the remnants had no effect on the weaker efflux produced by apoAI-deficient serum. Western blot analysis showed that granule remnants degrade apoAI in serum and in internal fluid. When released from remnants, chymase lost its ability to proteolyze HDL3 in the presence of serum. Thus, remnant chymase (but not isolated chymase) was able to resist the natural protease inhibitors present in serum and in intimal fluid. The results imply participation of exocytosed mast cell granules in foam cell formation in atherogenesis.

Animals↗
PubMed1996

Isolation and characterization of Caenorhabditis elegans extracellular matrix.

The extracellular matrix (ECM) plays an important structural and functional role in multicellular organisms. Because of the similarities between C. elegans and vertebrate development, this nematode could serve as a simplified model to study the biology of the ECM. In this study, a method for extracting mammalian ECM was adapted for the extraction of C. elegans ECM. ECM components from C. elegans were found to be homologous to mammalian ECM by immunoblotting. It was also demonstrated that antibodies generated against C. elegans ECM stained basement membrane-like structures in C. elegans eggs, larvae, and adults.

Animals↗
PubMed1996

Absence of association or genetic linkage between the angiotensin-converting-enzyme gene and left ventricular mass.

BACKGROUND: Homozygous carries of the D allele of the angiotensin-converting-enzyme (ACE) gene have been reported to be at increased risk for various cardiovascular disorders, including left ventricular hypertrophy. We investigated the potential role of the ACE gene in influencing left ventricular mass. METHODS: Quantitative echocardiographic data and DNA samples were available for 2439 subjects from the Framingham Heart Study. ACE genotypes were determined by an assay based on the polymerase chain reaction. (The D allele of the ACE gene contains a deletion, whereas the I [insertion] allele does not.) Left ventricular mass and the prevalence of left ventricular hypertrophy, adjusted for clinical covariates, were analyzed according to genotype. Genetic linkage between the ACE locus and left ventricular mass was evaluated by quantitative analysis of pairs of siblings. RESULTS: The ACE genotype was associated neither with left ventricular mass nor with the prevalence of left ventricular hypertrophy. Mean (+/-SE) left ventricular mass (adjusted for sex) among subjects carrying the DD, DI, and II genotypes was 165+/-1.6, 165+/-1.3, and 166+/-2.0 g, respectively (P=0.90). The prevalence of left ventricular hypertrophy among the three genotype groups was 15.6 percent, 13.6 percent, and 15.6 percent, respectively (P=0.36), and the adjusted relative risk of left ventricular hypertrophy associated with the DD genotype was 1.10 (95 percent confidence interval, 0.86 to 1.19). Linkage analysis in 759 pairs of siblings using both the ACE D/I marker and a microsatellite polymorphism at the neighboring locus for the human growth hormone gene failed to support any role of ACE in influencing left ventricular mass. CONCLUSIONS: The ACE genotype showed no association with echocardiographically determined left ventricular mass, nor did it confer an increased risk of left ventricular hypertrophy. We found no appreciable role of the ACE gene in influencing left ventricular mass.

Adult↗
PubMed1996

Association of fibrinogen and coagulation factors VII and VIII with cardiovascular risk factors in the elderly: the Cardiovascular Health Study. Cardiovascular Health Study Investigators.

The cross-sectional correlates of three hemostatic factors--fibrinogen, factor VII, and factor VIII--were examined in the Cardiovascular Health Study, a population-based cohort study of 5,201 subjects over age 65 years. Subjects were recruited in 1989-1990 in Forsyth County, North Carolina; Sacramento County, California; Washington County, Maryland; and Pittsburgh, Pennsylvania. In multivariate linear regression models, cardiac risk factors significantly associated with fibrinogen were current smoking, race, lipids, and white blood count. In women, alcohol use, obesity, physical activity, and insulin level were also significant, while in men hypertension was correlated. The significant correlates of factor VII were lipids and white blood count in men and estrogen use, alcohol use, race, lipids, insulin level, white blood count, and obesity in women. The independent correlates of factor VIII were insulin, glucose, and race in both sexes; low density lipoprotein cholesterol, white blood count, and diuretic use in men; and alcohol use in women. In multivariate models, factors known to be modifiable risk factors for cardiovascular disease accounted for more of the population variance of these hemostatic factors in women than in men, especially for factor VII. The hemostatic factors may mediate some effects of risk factors on disease, and this should be considered in longitudinal studies.

Age Distribution↗
PubMed1996

Down-regulation of protein kinase C in murine splenocytes: a potential mechanism for 2-acetylaminofluorene-mediated immunosuppression.

2-Acetylaminofluorene (AAF), an arylamide carcinogen, inhibited in a dose dependent manner mouse spleen cell proliferation in response to lipopolysaccharide (LPS). The objective of the present studies was to investigate the effects of AAF on protein kinase C (PKC) activation, an enzyme required for LPS-induced splenocytes proliferation. After treatment with 50 microM AAF for 18 h, PKC activity in the cytosolic fraction decreases by 50% from the control level, and splenocytes lost 30% of total PKC activity. Furthermore, as determined by the electrophoretic mobility shift assay, AAF inhibited the binding activity of the transcription factor complex, NF-KB, whose LPS-mediated induction is dependent on PKC activation in murine splenocytes. These results strongly suggest that LPS-mediated signaling in spleen cells is interrupted by AAF early in the signal transduction pathway, at a point proximal to the activation of PKC.

2-Acetylaminofluorene↗
PubMed1996

Intersubunit communication in tryptophan synthase by carbon-13 and fluorine-19 REDOR NMR.

The beta subunits of the 143-kDa alpha2beta2 tetrameric enzyme tryptophan synthase have been labeled by L-[ring-4-19F]phenylalanine and L-[phenol-4-13C]tyrosine in an effort to monitor the positions of these residues on ligand binding. Of the 13 phenylalanine and 11 tyrosine residues in the beta subunit, only three pairs have labels with 13C-19F separations of less than 6 angstrom. The beta subunit residues Tyr279 and Phe280 (each members of one of the three Tyr-Phe proximate pairs) have been suggested as possible conformational gates on ligand binding. The 188-MHz 19F NMR spectrum of the microcrystalline, double-labeled enzyme complex has five resolved lines under 5-kHz magic-angle spinning and 80-kHz proton dipolar decoupling. The distribution of beta-subunit 19F isotropic shifts is altered by addition of L-[3-13C]-serine to the mother liquor in contact with the microcrystals, consistent with a conformational rearrangement. The 13C label from serine is detected at 28 ppm as a methyl tautomer of bound aminoacrylate. The change in aromatic 19F chemical shifts on binding of serine indicates an alteration in local electric field gradients within the beta subunit. However, rotational-echo double-resonance 13C NMR (with 19F dephasing) shows that the average 13C-19F distance for the three phenylalanine-tyrosine proximate pairs in the beta subunit is changed by less than 1 angstrom.

Carbon Isotopes↗
PubMed1996

Structural variation of the pseudoautosomal region between and within inbred mouse strains.

The pseudoautosomal region (PAR) is a segment of shared homology between the sex chromosomes. Here we report additional probes for this region of the mouse genome. Genetic and fluorescence in situ hybridization analyses indicate that one probe, PAR-4, hybridizes to the pseudoautosomal telomere and a minor locus at the telomere of chromosome 9 and that a PCR assay based on the PAR-4 sequence amplifies only the pseudoautosomal locus (DXYHgu1). The region detected by PAR-4 is structurally unstable; it shows polymorphism both between mouse strains and between animals of the same inbred strain, which implies an unusually high mutation rate. Variation occurs in the region adjacent to a (TTAGGG)n array. Two pseudoautosomal probes can also hybridize to the distal telomeres of chromosomes 9 and 13, and all three telomeres contain DXYMov15. The similarity between these telomeres may reflect ancestral telomere-telomere exchange.

Animals↗
PubMed1996

Expression of functional glucagon receptors on lymphoid cells.

The objective of the present studies was to determine whether the existence of functional glucagon receptors could be established on lympoid cells. The glucagon receptor, which positively regulates adenylate cyclase, is a member of the superfamily of seven transmembrane domain G-protein coupled receptors. Previously reported specific binding with [125I]-glucagon to a variety of lymphoid and myeloid cell preparations suggests that glucagon receptors are expressed within the immune system. In the present study, Northern analysis of polyA RNA isolated from primary mouse and rat derived lymphoid tissues and lymphoid cell lines EL-4.IL-2, Jurkat E6-1, CH12LX, and BCL1-3B3 cells were probed with a 32P-labeled human hepatic glucagon receptor. Mouse spleen and thymus, rat spleen, and the B cell line, CH12LX, all possessed a single 1.5 kb fragment (BCL1-3B3, 1.4 kb) which hybridized to the glucagon receptor cDNA probe, as compared to mouse liver which exhibited a 2.8 kb fragment. EL-4.IL-2 and Jurkat E6-1 cells possessed a 3.7 kb fragment with an additional 2.75 kb band present in Jurkat E6-1 cells. Treatment of mouse splenocytes and T- and B-lymphoma cells with glucagon (0 - 100 nM) produced a dose-dependent enhancement in intracellular cAMP which was maximal at 5 min post treatment followed by a gradual decline. Direct addition of glucagon to spleen cell cultures over a broad concentration range produced no effect on either lymphoproliferation following stimulation with anti-CD3 mAb, or LPS nor on the antibody forming cell (AFC) response to sRBC. Conversely, glucagon effectively reversed the suppression of the sRBC AFC response produced by delta9-tetrahydocannabinol (delta9-THC), and partially reversed the suppression produced by 2',3'-dideoxyadenosine, both of which are potent inhibitors of adenylate cyclase. These studies confirm the expression of functional glucagon receptors on lymphoid cells.

Adenylyl Cyclases↗
PubMed1996

A wheelchair cushion designed to redistribute sites of sitting pressure.

OBJECTIVES: Despite the diversity of wheelchair cushions currently in clinical use, pressure on bony prominences continues to be a major problem for wheelchair-bound patients, and the incidence of pressure ulcers remains high. No static surface has been reported to reduce resting pressure under the ischial tuberosities to below that of capillary pressure, which may well be the threshold for inducing tissue damage. An entirely new form of seating was designed to decrease absolute pressure using a prosthetic fitting technique analogous to a below-the-knee prosthesis. DESIGN: A repeated measures randomized design was used to test differences between the experimental (TCS) and three other standard cushions. SETTING: A Veterans Medical Center outpatient service. PATIENTS: Wheelchair-bound volunteer subjects, n = 47, were selected who weighed more than 60kg. MAIN OUTCOME MEASURES: Pressures were measured by a standard air pressure pad and also by a computer-linked array of pressure transducers. The grid was standardized and used to generate topographic maps for each site over time. These data were used to measure the seating interface pressures. RESULTS: There was a significant main effect of cushion over site, F = 131 for left ischial tuberosity. Pressure were lowest while patients sat on the experimental seat and differences were significant at all time points. Using 1psi as presumed capillary pressure, frequency of success at achieving this pressure threshold was greatest for the experimental seat, p < .001. This difference persisted throughout the 30 minutes of testing. CONCLUSIONS: A computerized pressure grid was developed that allowed evaluation of anatomically localized pressures. The prosthetically designed TCS displays lower seating pressures than any other cushion tested. Consistent and sustained pressures were below the postulated threshold for tissue damage.

Adult↗
PubMed1996

In vitro model to evaluate the relative efficacy of catheter-directed thrombolytic strategies.

RATIONALE AND OBJECTIVES: Catheter-directed thrombolytic therapy has become an accepted treatment for many vascular occlusions. However, the relative rates of lysis of the different methods of drug administration have not been quantified. We developed an in vitro model to simulate and quantify local lytic therapy of a thrombotic vascular occlusion and tested it by evaluating three catheter-directed lytic strategies. METHODS: Seven-centimeter-long segments of 125I-fibrinogen-labeled thrombus made from recently expired human blood from a blood bank were formed in plastic tubes and were placed in a flowing stream of saline. Using multisidehole catheters, the clots were "treated" with intrathrombic saline or urokinase administered by drip infusion or forced injection using identical total doses of drug and volumes of fluid. Using endhole catheters, saline or urokinase was drip infused into the leading edge of the thrombus using the same protocol. A collimated scintillation detector was used to quantify the amount of activity remaining in the thrombus during each experiment, and the resultant time-activity curves for the different trials were compared. RESULTS: Forced-injection administration of urokinase using a multisidehole catheter produced the fastest lysis, resulting in a half-life of 42 min. The other infusion methods were slower, with half-lives of 153 min for multisidehole urokinase drip infusion, 365 min for endhole urokinase drip infusion, and more than 1,000 min for multisidehole catheter forced injection of saline and multisidehole and endhole saline drip infusion. The differences among these groups were reproducible and statistically significant. CONCLUSION: Results suggest that a simple and inexpensive in vitro model simulating lysis of a vascular occlusion can produce reproducible quantitative data. The data demonstrate that forced injection of lytic agents with a multisidehole catheter enhances the rate of thrombolysis and that the enhancement is not primarily attributable to the mechanical effect of this mode of administration.

Catheterization, Peripheral↗
PubMed1996

Clinical laboratory tests: application to daily practice.

Along with physical examination and medical history, clinical laboratory tests are an essential evaluative tool of health professionals. Pharmacists have a key role in selecting appropriate laboratory tests for TDM, interpreting tests to determine whether a drug is producing its desired therapeutic effect, and avoiding adverse drug effects. Prudence dictates that pharmacists should recommend that a laboratory test be done only if the results may lead to some action such as a dosage reduction, discontinuation of a drug, or ordering of additional tests to confirm the presence of an adverse effect. In these cost-conscious times, panels of laboratory tests that include multiple assays (Table 5) should be recommended only when it is less expensive to order the panel than to order individual tests, or when multiple tests are needed to evaluate the patient's situation. Pharmacists should be familiar with useful references to help interpret and apply lab test results to particular patient situations, so that they can make appropriate recommendations for altering drug therapy. Pharmacists with an appreciation of the role of clinical laboratory testing in patient care can perform a valuable service for their patients and other health professionals.

Clinical Laboratory Techniques↗