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Biomedical subjects

J M Anderson

Publications and source records attributed to J M Anderson.

At least 19 recordsLinked to original sources
PubMed2000

hCASK and hDlg associate in epithelia, and their src homology 3 and guanylate kinase domains participate in both intramolecular and intermolecular interactions.

Membrane-associated guanylate kinase (MAGUK) proteins act as molecular scaffolds organizing multiprotein complexes at specialized regions of the plasma membrane. All MAGUKs contain a Src homology 3 (SH3) domain and a region homologous to yeast guanylate kinase (GUK). We showed previously that one MAGUK protein, human CASK (hCASK), is widely expressed and associated with epithelial basolateral plasma membranes. We now report that hCASK binds another MAGUK, human discs large (hDlg). Immunofluorescence microscopy demonstrates that hCASK and hDlg colocalize at basolateral membranes of epithelial cells in small and large intestine. These proteins co-precipitate from lysates of an intestinal cell line, Caco-2. The GUK domain of hCASK binds the SH3 domain of hDlg in both yeast two-hybrid and fusion protein binding assays, and it is required for interaction with hDlg in transfected HEK293 cells. In addition, the SH3 and GUK domains of each protein participate in intramolecular binding that in vitro predominates over intermolecular binding. The SH3 and GUK domains of human p55 display the same interactions in yeast two-hybrid assays as those of hCASK. Not all SH3-GUK interactions among these MAGUKs are permissible, however, implying specificity to SH3-GUK interactions in vivo. These results suggest MAGUK scaffold assembly may be regulated through effects on intramolecular SH3-GUK binding.

Binding Sites↗
PubMed2000

Noninvasive assessment of reperfusion after fibrinolytic therapy for acute myocardial infarction.

Assessment of reperfusion by the 12-lead electrocardiogram (ECG) or biochemical markers is limited by suboptimal sensitivity and/or specificity. Body surface mapping (BSM) improves the spatial sampling of the 12-lead ECG. Serial 12-lead ECGs and 64-lead anterior BSMs were recorded from 67 patients with acute myocardial infarction undergoing coronary angiography 90 minutes after fibrinolytic therapy. ECG-1 and BSM-1 were recorded before/shortly after therapy (median 18 minutes). ECG-2 and BSM-2 were recorded after the 90-minute angiogram (median 30 minutes). The maximum ST elevation on ECG-1 was noted and > or = 30% ST resolution on ECG-2 was taken to represent partial/complete reperfusion. Patients were randomly divided into a training set and validation set. Isointegral and isopotential ST-T variables from BSMs of training-set patients were compared with Thrombolysis In Myocardial Infarction (TIMI) trial flow using discriminant analysis to identify which variables best classified reperfusion. Reperfusion (TIMI 2/3 flow) occurred in 32 of 34 training-set patients and in 29 of 33 validation-set patients. In the training set, > or = 30% ST resolution correctly classified reperfusion with 72% sensitivity (23 of 32) and 50% specificity (1 of 2). In the validation set, > or = 30% ST resolution classified reperfusion with 59% sensitivity (17 of 29) and 50% specificity (2 of 4). In comparison, a model containing 24 BSM variables correctly classified all training-set patients, and when prospectively tested in the validation-set, correctly classified 28 of 29 patients who achieved reperfusion (97% sensitivity) and all 4 patients who failed to reperfuse (p = 0.035). In conclusion, BSM is more useful than the 12-lead ECG for noninvasive assessment of reperfusion after fibrinolytic therapy for acute myocardial infarction.

Analysis of Variance↗
PubMed2000

Increased discrimination of "false memories" in autism spectrum disorder.

Individuals with autism spectrum disorder (ASD) have impaired ability to use context, which may manifest as alterations of relatedness within the semantic network. However, impairment in context use may be more difficult to detect in high-functioning adults with ASD. To test context use in this population, we examined the influence of context on memory by using the "false memory" test. In the false memory task, lists of words were presented to high-functioning subjects with ASD and matched controls. Each list consists of words highly related to an index word not on the list. Subjects are then given a recognition test. Positive responses to the index words represent false memories. We found that individuals with ASD are able to discriminate false memory items from true items significantly better than are control subjects. Memory in patients with ASD may be more accurate than in normal individuals under certain conditions. These results also suggest that semantic representations comprise a less distributed network in high-functioning adults with ASD. Furthermore, these results may be related to the unusually high memory capacities found in some individuals with ASD. Research directed at defining the range of tasks performed superiorly by high-functioning individuals with ASD will be important for optimal vocational rehabilitation.

Adult↗
PubMed2000

CaCo-2 cells treated with Clostridium perfringens enterotoxin form multiple large complex species, one of which contains the tight junction protein occludin.

The previous model for the action of Clostridium perfringens enterotoxin (CPE) proposed that (i) CPE binds to host cell receptor(s), forming a small ( approximately 90 kDa) complex, (ii) the small complex interacts with other eucaryotic protein(s), forming a large ( approximately 160 kDa) complex, and (iii) the large complex triggers massive permeability changes, thereby inducing enterocyte death. In the current study, Western immunoblot analysis demonstrated that CPE bound to CaCo-2 human intestinal cells at 37 degrees C forms multiple large complex species, with apparent sizes of approximately 200, approximately 155, and approximately 135 kDa. These immunoblot experiments also revealed that occludin, an approximately 65-kDa tight junction protein, is present in the approximately 200-kDa large complex but absent from the other large complex species. Immunoprecipitation studies confirmed that occludin physically associates with CPE in large complex material and also indicated that occludin is absent from small complex. These results strongly suggest that occludin becomes associated with CPE during formation of the approximately 200-kDa large complex. A postbinding association between CPE and occludin is consistent with the failure of rat fibroblast transfectants expressing occludin to bind CPE in the current study. Those occludin transfectants were also insensitive to CPE, strongly suggesting that occludin expression is not sufficient to confer CPE sensitivity. However, the occludin-containing, approximately 200-kDa large complex may contribute to CPE-induced cytotoxicity, because nontoxic CPE point mutants did not form any large complex species. By showing that large complex material is comprised of several species (one containing occludin), the current studies indicate that CPE action is more complicated than previously appreciated and also provide additional evidence for CPE interactions with tight junction proteins, which could be important for CPE-induced pathophysiology.

Animals↗
PubMed2000

Shear stress-induced apoptosis of adherent neutrophils: a mechanism for persistence of cardiovascular device infections.

The mechanisms underlying problematic cardiovascular device-associated infections are not understood. Because the outcome of the acute response to infection is largely dependent on the function of neutrophils, the persistence of these infections suggests that neutrophil function may be compromised because of cellular responses to shear stress. A rotating disk system was used to generate physiologically relevant shear stress levels (0-18 dynes/cm(2); 1 dyne = 10 microN) at the surface of a polyetherurethane urea film. We demonstrate that shear stress diminishes phagocytic ability in neutrophils adherent to a cardiovascular device material, and causes morphological and biochemical alterations that are consistent with those described for apoptosis. Complete neutrophil apoptosis occurred at shear stress levels above 6 dynes/cm(2) after only 1 h. Morphologically, these cells displayed irreversible cytoplasmic and nuclear condensation while maintaining intact membranes. Analysis of neutrophil area and filamentous actin content demonstrated concomitant decreases in both cell area and actin content with increasing levels of shear stress. Neutrophil phagocytosis of adherent bacteria decreased with increasing shear stress. Biochemical alterations included membrane phosphatidylserine exposure and DNA fragmentation, as evaluated by in situ annexin V and terminal deoxynucleotidyltransferase-mediated dUTP end labeling (TUNEL) assays, respectively. The potency of the shear-stress effect was emphasized by comparative inductive studies with adherent neutrophils under static conditions. The combination of tumor necrosis factor-alpha and cycloheximide was ineffective in inducing >21% apoptosis after 3 h. These findings suggest a mechanism through which shear stress plays an important role in the development of bacterial infections at the sites of cardiovascular device implantation.

Actins↗
PubMed2000

Adsorbed IgG: a potent adhesive substrate for human macrophages.

Previous reports from our laboratory have demonstrated qualitatively that preabsorbed IgG can enhance long-term macrophage adhesion in vitro. This investigation further characterizes and quantifies the biological effect of adsorbed human IgG on human macrophages and probes the potential mechanisms. Ten-day human monocyte/macrophage cultures on Plastek M (PM), a normally poor cellular substrate for macrophages, confirmed the ability of preabsorbed IgG to dramatically enhance long-term macrophage adhesion. An adsorption solution concentration of 200 microg/mL of IgG was necessary to provide a consistent, optimal cellular response. (125)I adsorption studies indicated Langmuir-style IgG adsorption at low concentrations; however, no adsorption maximum was observed. Additional adsorption analysis revealed that the IgG fragments Fab, F(ab')(2), and Fc adsorb at levels only 20-40% that of the whole molecule. Despite the lower adsorption levels, both preabsorbed Fab and F(ab')(2) were shown to be as effective as whole molecule IgG at enhancing long-term macrophage adhesion. Surprisingly, the preabsorbed Fc fragment demonstrated no IgG-like activity, thereby eliminating the possibility of an Fc receptor-based mechanism. Other possible mechanisms, such as macrophage lectins, novel macrophage Fab receptors, and complement activation by adsorbed IgG and IgG fragments, are discussed.

Biocompatible Materials↗
PubMed2000

Early diagnosis of right ventricular or posterior infarction associated with inferior wall left ventricular acute myocardial infarction.

Right ventricular (RV) or posterior infarction associated with inferior wall left ventricular acute myocardial infarction (AMI) has important therapeutic and prognostic implications. However, RV and posterior chest leads in addition to the 12-lead electrocardiogram are required for accurate detection. Body surface mapping (BSM) has greater spatial sampling and may further improve inferior wall AMI classification. Consecutive patients with chest pain lasting <12 hours were assessed to identify those with AMI and > or =0.1 mV ST elevation in > or =2 contiguous inferior leads of the 12-lead electrocardiogram (bundle branch block or left ventricular hypertrophy excluded). A 12-lead electrocardiogram, RV leads (V(2)R, V(4)R), posterior chest leads (V(7), V(9)), and a BSM were recorded. From each BSM, the 12 electrodes overlying the RV region (regional RV map) and 10 electrodes overlying the posterior wall (regional posterior map) were assessed for ST elevation. Infarct size was estimated by serial cardiac enzymes. AMI occurred in 173 of 479 patients. Of the 62 patients with inferior wall AMI, ST elevation > or =0.1 mV occurred in 26 patients (42 in V(2)R or V(4)R compared with 36 patients (58%) in > or =1 electrode on the regional RV map (p = 0.0019). ST elevation > or =0.1 mV occurred in 1 patient (2%) in V(7) or V(9) compared with 17 patients (27%) in > or =1 electrode on the regional posterior map (p = 0.00003). ST elevation > or =0.05 mV occurred in 6 patients (10%) in V(7) or V(9) compared with 22 patients (36%) in > or =1 electrode on the regional posterior map (p = 0.00003). Patients with ST elevation on regional RV and/or posterior maps had a trend toward larger infarct size (mean peak creatine kinase 1,789+/-226 vs. 1,546+/-392 mmol/L; p = NS). Thus, BSM, when compared with RV or posterior chest leads, provides improved classification of patients with inferior wall AMI and RV or posterior wall involvement.

Body Surface Potential Mapping↗
PubMed2000

Adsorbed serum proteins responsible for surface dependent human macrophage behavior.

Substrate specific cellular responses are the result of a complex biological system that includes protein adsorption, receptor-ligand binding, and signal transduction. This investigation attempted to identify specific proteins adsorbed from human serum that may be responsible for the previously reported in vitro surface dependent behavior of human macrophages and foreign body giant cells (FBGCs). The adsorption of human albumin, alpha(2)-macroglobulin, complement factor 3b, fibronectin, IgG, thrombospondin, vitronectin (VN), and von Willebrand factor (vWF) from a 25% serum solution was quantified with (125)I-labeled protein. Adsorption substrates included clean glass, alkyl-silane modified glass, amino-silane modified glass, poly(ethylene oxide) (PEO)-coupled glass, and the reference biomaterials poly(etherurethane urea), Silastic(R), and poly(tetrafluoroethylene) (PTFE). Following quantification of 2-h adsorption, surfaces were treated with sodium dodecyl sulfate (SDS) and the level of adsorbed proteins remaining was quantified. The pre- and post-SDS adsorption were both compared to previously reported surface dependent in vitro macrophage and FBGC behavior on the same surfaces; however, no correlations could be made. Adsorption strength, defined as the percentage of initially adsorbed protein that remained adsorbed after SDS treatment, correlated well with previously reported in vitro cellular behavior indicating that adsorbed vWF, IgG, and VN may be involved in the modulation of adherent macrophage and FBGC behavior. Those surfaces that strongly adsorbed vWF also inhibited long-term macrophage adhesion, while those surfaces that strongly adsorbed IgG promoted long-term macrophage adhesion. In addition, the highest levels of FBGC formation had been observed only on those surfaces that strongly adsorbed VN. Subsequent human monocyte cultures on protein preadsorbed substrates confirmed the inhibitory effect of adsorbed vWF and the promoting effect of IgG on longterm macrophage adhesion as predicted by adsorption strength correlations. However, preadsorbed VN was not observed to modulate FBGC formation, which is in contrast to the conclusions of the adsorption correlations.

Adsorption↗
PubMed2000

High-dose-rate postoperative vaginal cuff irradiation alone for stage IB and IC endometrial cancer.

PURPOSE: To evaluate the effectiveness of postoperative high-dose-rate (HDR) vaginal cuff irradiation alone (1500 cGy in 3 fractions) in patients with Stage Ib and Ic endometrial cancer. METHODS AND MATERIALS: This is a retrospective review of 102 patients with Stage Ib and Ic endometrial cancer treated with a hysterectomy and postoperative HDR intracavitary therapy alone during the period of 1/1/90-12/31/96. Each patient received 1500 cGy in 3 weekly treatments, dosed to a depth of 0.5 cm. Pathologic features such as depth of invasion, tumor grade, lower uterine segment (LUS) involvement, and lymphvascular invasion (LVI) were evaluated for their impact on recommended postoperative treatment. All survival curves were generated utilizing Kaplan-Meier methods and all statistical comparisons were via a Wilcoxon rank sum test. RESULTS: The 5-year actuarial overall survival (OS) is 84% and the 5-year disease-free survival (DFS) is 93%. Locoregional disease control (pelvic control) was excellent with 97% of the patients free of pelvic disease at 5 years. Of the three pelvic failures only one was in the vaginal cuff. LVI, LUS involvement, Grade 3 and/or outer third myometrial involvement were identified in 41 patients. Thirty-one of these patients underwent a lymphadenectomy and there were two regional failures within this increased-risk group. CONCLUSIONS: We obtained an excellent level of locoregional control with minimal morbidity and minimal time commitment for treatment with vaginal HDR brachytherapy alone. Our dose per fraction and total dose is lower than most reported series and there is no apparent loss in locoregional control. In addition, intermediate-risk patients and patients with an increased risk of recurrence (Grade 3, outer third myometrial involvement, LVI, LUS) may be treated with cuff irradiation alone, after surgical staging and a negative lymphadenectomy.

Adenocarcinoma↗
PubMed2000

High molecular weight kininogen inhibition of endothelial cell function on biomaterials.

Synthetic vascular grafts implanted into humans fail to develop a complete endothelial lining. In previous studies, we have shown that high-molecular-weight kininogens (HMWK) adsorb to the surfaces of biomaterials. In addition, it has been demonstrated that these proteins modulate cellular function. In the present study, we report on the adhesion and proliferation of human umbilical-vein endothelial cells (HUVEC) on tissue culture polystyrene, glass, polyurethane, and Mylar(trade mark) surfaces coated with human HMWK, either single-chain HMWK (SC-HMWK) or double-chain HMWK (DC-HMWK). Surfaces coated with fibronectin served as a positive control for these experiments. Parallel experiments were performed in which HUVEC were allowed to migrate from crosslinked dextran microcarrier beads (Cytodex 2) onto HMWK-coated surfaces. Our results indicate that HMWK-coated surfaces inhibit endothelial cell adhesion, proliferation, and migration at 24 and 72 h, and this inhibition is concentration dependent. To determine a potential mechanism for this inhibitory phenomenon, cells were stained for cytoskeletal actin filaments using rhodamine-phalloidin. Endothelial cells on HMWK-coated surfaces displayed F-actin filament reorganization/disassembly, characterized by the absence of peripheral actin bands in focal adhesion contacts. We conclude that HMWK inhibit endothelial cell adhesion, proliferation, and migration on a variety of biomaterial surfaces. This inhibitory effect may play a role in promoting the lack of endothelialization in synthetic vascular grafts, which is thought to play a significant role in the failure of these devices.

Adsorption↗
PubMed2000

Prevention of monocyte adhesion and inflammatory cytokine production during blood platelet storage: an in vitro model with implications for transfusion practice.

A novel platelet additive solution [ThromboSoltrade mark (TS)] was designed to allow extended refrigerated platelet storage. It has been shown to preserve platelet function and prevent cytokine accumulation in platelet concentrates stored for up to 9 days. It consists of amiloride, adenosine, sodium nitroprusside, dipyridamole, quinacrine, and ticlopidine. We hypothesized that the cytokine inhibition may be due to prevention of monocyte (MC) adhesion and activation on the surfaces of platelet storage bag plastic polymers. In an in vitro model, we incubated purified peripheral blood MCs on discs of polyolefin and polyvinylchloride from platelet storage bags, and on polystyrene, in the presence of TS for up to 7 days. We found that after incubation with TS, adherent MC numbers were decreased by >80-95% compared with controls on all surfaces examined. Levels of cytokines [interleukin (IL)-1beta, IL-1RA, IL-6, IL-8, and tumor necrosis factor-alpha] were low in wells with TS but rose progressively in the controls during incubation. Amiloride alone had similar effects on adhesion and cytokine release as the complete TS preparation. Removing amiloride from TS abrogated these effects. These findings suggest an important role for TS and amiloride in monocyte function, and have implications for the development of agents designed for prolonged platelet storage.

Blood Platelets↗
PubMed2000

Incorporation of (15)N from spiked cattle dung pats into soil under two moorland plant communities.

The rate and depth of cattle dung incorporation into moorland soil may be an important factor influencing plant community dynamics through its effects on soil nutrient availability. This study traces the incorporation of (15)N-labelled dung into a moorland soil under two vegetation types in Dartmoor National Park, UK. Cores of treated and control soil 10 cm deep were collected at 2, 4, 8 and 16 week intervals and divided into 2 cm depth increments. Soil samples were freeze-dried, ground and analysed for atom% (15)N and %N content using continuous-flow isotope-ratio mass spectrometry. The contribution of dung N to the soil N pool was estimated by changes in atom% (15)N of the soil. The incorporation of dung dry matter into the soil was also calculated. The labile component of the dung N was incorporated deeper and more rapidly into soil under grass than under heather vegetation. The implications of these processes for the dynamics of upland plant communities are considered in relation to the ability of plants to compete for nutrients.

Animals↗
PubMed2000

Laboratory-scale mass production of a multi-micropatterned grafted surface with different polymer regions.

In this article, we demonstrate laboratory-scale mass production of a regionally precise multi-micropatterned surface photo-graft-copolymerized with three water-soluble monomers based on the photochemistry of an iniferter, which means that it acts as an initiator, a transfer agent and a terminator, benzyl N, N-diethyldithiocarbamate. The surface was semi-automatically prepared using a combination of a custom-designed irradiation apparatus installed with a motor-controlled stage for a substrate and three photomasks with different line-patterned slits (number of slits 20, width 500 microm, length 10 mm), and carbon dioxide laser cutting apparatus. A particular region of poly(styrene-co-vinylbenzyl N,N-diethyldithiocarbamate) coated on a PET film was irradiated in a particular aqueous monomer solution while moving the irradiated portion stepwise after irradiation through each line of the photomask. Photo-graft-copolymerization was carried out sequentially with acrylic acid sodium salt (AANa), N-[3-(dimethylamino)propyl]acrylamide methiodide (DMAPAAm), and acrylamide (AAm) using differently patterned photomasks. Characterization of surface elemental distribution by X-ray photoelectron spectroscopy (XPS), and light microscopic visualization by dye staining revealed a microprocessed surface with 20 sets of micropatterns, each of which had three line regions grafted with three different polymers. The irradiation of a carbon dioxide laser manipulated via computer-aided design (CAD) software onto the microprocessed surface resulted in automatic circular cutting for each set of micropatterns to mass-produce multi-micropatterned substrates for the study of substrate-dependent endothelial cell responses.

Animals↗
PubMed2000

Improved biocompatibility of a viscous bioerodible poly(ortho ester) by controlling the environmental pH during degradation.

The poly(ortho ester), POE, used in this investigation, is a viscous bioerodible polymer (8 kDa), which rapidly degrades into a triol and an acidic by-product, acetic acid. In order to improve biocompatibility, we have evaluated the addition of various basic excipients, such as sodium acetate, hydroxyapatite, calcium carbonate and magnesium hydroxide, which buffered and neutralized the acidic degradation product and prolonged the polymer lifetime and drug release. This decrease of POE degradation rate results in a decreased rate of formation of the acidic by-product. Similarly, a POE of higher molecular weight (14 kDa) has been tested. Sodium acetate was too hydrophilic to affect the drug release and the biocompatibility of the polymer, whereas the presence of magnesium hydroxide markedly prolonged the drug release and improved the acceptability of the polymer. The increased molecular weight POE did not improve biocompatibility and a similar but delayed, inflammatory reaction was observed.

Animals↗
PubMed2000

Changes in lipoprotein(a) levels measured by six kit methods during growth hormone treatment of growth hormone-deficient adults.

Lipoprotein(a) [Lp(a)], an independent risk factor for cardiovascular disease, has previously been reported to increase, decrease or show no change in growth hormone (GH)-deficient individuals receiving GH replacement. To assess whether these inconsistencies could be attributed to differences in immunoassay methods, Lp(a) was measured by six commercial kits at 0, 3, 6 and 9 months in nine GH-deficient individuals (median age 68.3 years, six male) during 9 months GH therapy. There was a significant rise in Lp(a) with the INCStar immunoturbidimetric (IT) method and the Mercodia enzyme linked immunosorbent assay (ELISA) (P</=0.05, two-tailed Wilcoxon signed rank test), a non-significant rise with the Pharmacia immuno-radiometric assay and the Biopool ELISA methods (P =0.06), and no change with the Immuno ELISA and WAKO IT kits. There was also considerable variation in the values reported within each individual. These results suggest that the previously reported inconsistencies may in part be due to methodological differences, and that the effect of GH on Lp(a) remains unknown. This study highlights the need for a more common approach to the standardization of Lp(a) methods and the selection of antibodies used in them. Better performing methods may allow a more reliable interpretation of the effects of GH on Lp(a)

Aged↗