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Biomedical subjects

J M Anderson

Publications and source records attributed to J M Anderson.

At least 181 records · Page 10Linked to original sources

Complement C3 participation in monocyte adhesion to different surfaces.

As part of an ongoing investigation into the role of the monocyte/macrophage in biocompatibility, a major goal is to identify the adhesion mechanisms that initiate and promote the observed in vivo morphologic progression of monocyte-to-macrophage-to-foreign body giant cell on biomaterials. We have exploited differently modified polystyrenes, specific component-depleted sera, and monoclonal antibodies (mAbs) to leukocyte integrins to ask what adhesion mechanisms mediate human blood monocyte adhesion to different surfaces in vitro. Preliminary findings are that monocyte interactions with fluorinated, siliconized, nitrogenated, and oxygenated surfaces are reduced by 50-100% when complement component C3-depleted serum is used for adsorption; reductions vary with material surface properties. Adhesion is restored on all surfaces when C3-depleted serum is replenished with purified C3. Monocyte adhesion to serum-adsorbed surfaces is inhibited by mAbs to the leukocyte integrin beta subunit, CD18 (mAbs 60.3 and MHM23), and partially inhibited by a mAb to the alpha subunit, CD11b (mAb 60.1), suggesting adhesive interactions between adsorbed C3bi (the hemolytically inactive form of the C3b fragment) and the leukocyte integrin CD11b/CD18. However, adsorbed fibrinogen reduces the effectiveness of these mAbs, indicating that alternative adhesion mechanisms may operate depending on the propensities of critical adhesion-mediating components to be adsorbed onto different surfaces.

Antibodies, Monoclonal↗

Expression of cytokeratin confers multiple drug resistance.

The cytokeratin network is an extensive filamentous structure in the cytoplasm whose biological function(s) is unknown. Based upon previous data showing the modification of cytokeratin by mitoxantrone, we investigated the ability of cytokeratin networks to influence the survival response of cells to chemotherapeutic agents. We have compared the survival of mouse L fibroblasts lacking cytokeratins with that of L cells transfected with cytokeratins 8 and 18 in the presence of chemotherapeutic drugs. The expression of cytokeratins 8 and 18 conferred a multiple drug resistance phenotype on cells exposed to mitoxantrone, doxorubicin, methotrexate, melphalan, Colcemid, and vincristine. The degree of drug resistance was 5-454 times that of parental cells, depending upon the agent used. Drug resistance could not be attributed to altered growth characteristics, altered drug accumulation, or an altered drug efflux in the transfected cells. Cytokeratin does not confer resistance to ionizing radiation, which damages DNA independently of intracellular transport mechanisms. These data suggest a role for cytokeratin networks in conferring a drug resistance phenotype.

Animals↗

Theoretical analysis of in vivo macrophage adhesion and foreign body giant cell formation on polydimethylsiloxane, low density polyethylene, and polyetherurethanes.

Quantitative description of foreign body giant cell (FBGC) formation on implanted polymer surfaces as a function of time can conceivably correlate cell adhesion with polymer properties and possibly predict the behavior of the polymer in vivo. In the present study, the formation of FBGCs on various biomedical polymers was quantified by two parameters: the density of adherent macrophages present initially that participate in FBGC formation (d0) and the rate constant for cell fusion (k); both kinetic parameters were used to calculate the time-dependent FBGC density (dfc). The materials used were: three Pellethane poly(etherurethanes) (PEUs) varying in weight percent of hard segment, one poly(etherurethane urea) (PEUU), and NHLBI-DTB primary reference materials: low density polyethylene (LDPE), silica-free polydimethylsiloxane (PDMS). The results indicated that up to 5 weeks of implantation, FBGCs were formed from the fusion of one population of adherent macrophages present by 3 days post-implantation. Furthermore, only a small fraction (< 8%) of this initial adherent macrophage population participated in FBGC formation. Based on the results of previous studies and the current study, it was concluded that increase in PEU hard segment weight percent, surface hardness and hydrophobicity increased total protein adsorption and effectively increased d0 and dfc. No further correlations between the material properties of all polymers and the cell kinetics can be made at this time. However, this study demonstrated that macrophage adhesion and FBGC formation can be quantified with the cell fusion model, and are modulated by various polymer properties.

Adsorption↗

Creep of a poly(etherurethane urea) in an oxidative environment.

The creep behavior of a PEUU without added stabilizers was examined in H2O2/CoCl2, an environment that simulates the biodegradation of this polymer. Creep in the control environments, air, water, and H2O2, was logarithmic with time as is characteristic of primary or viscoelastic creep. At short times, creep in H2O2/CoCl2 followed the same time dependency as creep in H2O2; however, at longer times an acceleration in the creep rate was observed. Creep in H2O2/CoCl2 was satisfactorily described by addition of a linear time term to the creep equation with an induction time, ti. The induction time was extended by stress-induced crystallization of the soft segments, but was reduced by an increase in H2O2 concentration. Oxidative degradation of the PEUU soft segments was detected by infrared and GPC analysis at times less than ti. This led to the speculation that an initial "precursor" layer was created at the surface by chain cleavage. Microcracking in a subsequent stage was postulated to be responsible for the observed effect on the creep behavior.

Biocompatible Materials↗

Ion-selective membranes with low plasticizer content: electroanalytical characterization and biocompatibility studies.

High molecular weight poly(vinyl chloride) and aliphatic polyurethane (Tecoflex)-based ion selective membranes, with normal and reduced amounts of plasticizer, as well as without plasticizer, were tested with respect to their analytical properties, their biocompatibility, and cellular responses. The analytical properties of the membranes did not change significantly within a wide range of polymer to plasticizer ratios. However, the membranes with reduced plasticizer content had better adhesive properties, less anion interference, extended life time, and better biocompatibility. Using the cage implant system, the results showed that an increase of plasticizer weight percent in Tecoflex membranes correlated positively with the increase in host inflammatory response up to 14 days of implantation. The results also demonstrated that both PVC and Tecoflex-based ion-selective membranes with the most common membrane composition (1:2 polymer to plasticizer ratio) exhibited a similar acute inflammatory response, but the PVC-based membrane elicited a reduced chronic inflammatory response when compared with the Tecoflex-based membrane.

Adhesiveness↗

In vivo induction of macrophage Ia antigen (MHC class II) expression by biomedical polymers in the cage implant system.

Examination of the cellular components in the inflammatory exudate, which infiltrates subcutaneous cages, can be used to monitor the progress of an inflammatory response to an implanted material. Of particular interest is the study of monocyte/macrophage infiltration into the implanted cages containing biomaterials, as macrophages may initiate a wide spectrum of responses upon interaction with a foreign material. In this study, the authors propose a technique using subcutaneous tissue cages in conjunction with cytofluorimetric analysis of exudate leukocytes to evaluate the monocyte/macrophage cell activation in response to different materials. The studies reported here used several materials (thermoplastic and elastomeric polymers) as the challenging agent, to demonstrate whether polymers, chemically different from each other, could differentially activate macrophages to carry out their proinflammatory role more effectively. The materials tested included: poly(etherurethane ureas) (PEUU A'), poly(etherurethane ureas) with a surface active additive, Methacrol, (PEUU C'), polymethylsiloxane (PDMS), polyetherimide, (PEI), and polyetheretherketone, (PEEK). For all tested materials, the maximum numbers of exudate cells and of Ia-positive macrophages were found on day 7, although the entity of the cell increase was associated with the material used for the implant. Similarly, the percentage of Ia-positive macrophages varied according to the specific polymer present in the cages after 7 days. By day 14, the percentage of Ia-positive macrophages decreased with individual exudates showing 19-32% Ia-positive cells depending on the different type of material. Only in the case of PDMS did the percentage of Ia-positive macrophages remain the same as compared with control empty cage macrophages.

Animals↗

Theoretical analysis of in vivo macrophage adhesion and foreign body giant cell formation on strained poly(etherurethane urea) elastomers.

Quantitative description of foreign body giant cell (FBGC) formation on poly(etherurethane urea) (PEUU) surfaces as a function of time can conceivably predict the effects of polymer characteristics on cellular responses in vivo. In the present study, the formation of FBGCs on strained and unstrained PEUUs was quantified with two parameters: the density of adherent macrophages present initially that participate in FBGC formation (d(o)) and the rate constant for cell fusion (k); both kinetic parameters were used to calculate the time-dependent FBGC density (dfc). Relationships were sought between results of the cellular analysis and the extent of environmental stress cracking (ESC), as characterized by scanning electron microscopy. Surface degradation was semiquantified with percent light transmittance. The materials used were: base PEUU, base PEUU with 1% Santowhite antioxidant powder, base PEUU with 5% Methacrol 2138F antifume agent, and base PEUU with both 1% Santowhite and 5% Methacrol 2138F. A comparison of unstrained base PEUU with base PEUU strained to 400% elongation indicated that the rate of cell fusion, but not d(o) and dfc, increased in the presence of strain. In all strained samples, additives that strongly affected the ESC also influenced FBGC kinetic parameters. Strained PEUU containing Santowhite had the lowest d(o), the slowest rate of cell fusion, and lowest dfc, and the least incidence of ESC. The results suggest that the incidence of ESC in PEUU was decreased in the presence of Santowhite, which also lowered the number of adherent macrophages participating in FBGC formation, the rate of FBGC formation and the subsequent FBGC density. These studies also indicate that strain in PEUUs does not directly modulate the adherent macrophage and FBGC density. Further studies are necessary to delineate the relationship between PEUU strain and adherent macrophage and FBGC activation, which leads to the exocytosis of degrading agents and the observed incidence of biodegradation.

Animals↗

Biocompatibility of a new semisolid bioerodible poly(ortho ester) intended for the ocular delivery of 5-fluorouracil.

The biocompatibility of a new semisolid, hydrophobic poly(ortho ester) (POE) intended for controlled drug delivery to the eye was evaluated. The polymer was injected subconjunctivally in rabbits, and clinical and histologic examinations were performed 3, 10, 15, and 21 days after injection. Polymers injected as controls were an aqueous gel of sodium hyaluronate (SH), 1% in phosphate buffer, and medical grade silicone oil. After injection, the POE emulsified into small droplets and a focal eosinophilic reaction was noted at 3 days' implantation. At 10 days' implantation, the POE was not identified in the implantation site and the inflammatory reaction had resolved, with fibroblasts being the predominant cell type. At 15 and 21 days, no POE was identified and normal appearing tissue was present in the injection site. Sodium hyaluronate was not inflammatory over the period of the implantations. Silicone oil induced a slight inflammation at 3 days, with the presence of eosinophils and limited necrosis with cellular debris. Silicone oil was present in the implantation site at 3, 10, 15, and 21 days. The inflammatory response to the respective polymers was evaluated in the subconjunctival tissue. The inflammatory reaction was quantified at the implant site, adjacent subconjunctival tissues, and scleral and corneal stroma. The inflammatory cell densities in these respective tissue zones were determined, and the ratio of eosinophils over total inflammatory cells was calculated. POE did not become encapsulated with fibrous tissue, but biodegraded in a short time, indicating its potential for use after glaucoma filtration surgery.

Animals↗

Specificity of replicase-mediated resistance to cucumber mosaic virus.

Plants transformed with a nucleotide sequence coding for a truncated RNA 2 replicase gene of the subgroup I strain of cucumber mosaic virus, Fny-CMV, are resistant to cucumber mosaic disease. Two resistant lines representing independent transformations in the original study have been propagated, and their progeny have been examined. Resistance to Fny-CMV was genetically integrated and was retained in the R4 generation. Fourteen subgroup I CMV strains, 6 strains uncharacterized as to subgroup, and 4 subgroup II CMV strains, as well as potato virus X, tobacco mosaic virus, tomato aspermy virus, tobacco etch virus, and peanut stunt virus were inoculated mechanically to R2 generation plants of these two lines. Resistance was found to 12 subgroup I CMV strains and to all of the subgroup-uncharacterized strains. No resistance was found to subgroup II CMV strains nor to the other viruses listed above. However, the two plant lines behaved differently in response to 3 subgroup I strains, K-CMV, PRC-CMV, and MB-CMV, where one line (5) was more resistant than the other (2). RNase protection analyses indicated that these 3 strains vary more in sequence from Fny-CMV than do the other subgroup I strains tested. Resistance was also shown when aphids were used to inoculate plants with either of 2 subgroup I CMV strains; no resistance was observed when aphids were used to transmit a subgroup II CMV strain. Inoculation of transgenic plants with pseudorecombinants produced with the RNAs derived from Fny-CMV to which the plants are resistant, and LS-CMV, a subgroup II strain to which they are susceptible, indicated that the resistance-breaking capacity of LS-CMV is a function of RNA 2.

Animals↗

Problems and approaches in covalent attachment of peptides and proteins to inorganic surfaces for biosensor applications.

Some of the fundamental problems in covalent attachment of peptides and proteins to putative biosensor surfaces are reviewed and specific approaches to these problems discussed. In addition, selected aspects of our recent work utilizing self-assembled monolayer (SAM) systems designed to react selectively with the thiol side chain of Cys in proteins are presented. Uniform attachment of a 21-amino acid peptide antigen through a single Cys residue with retention of biological function (antibody binding) has been attained. Further work with this system may lead to solutions for some of the problems which currently prevent the development of reliable biosensors for industrial and medical use.

Amino Acid Sequence↗

Immunohistochemical localisation of hydroxysteroid sulphotransferase in human breast carcinoma tissue: a preliminary study.

Understanding the function and regulation of the metabolism of steroid hormones by breast tumours will be instrumental to the development of novel treatments for this widespread disease. We have examined the expression of hydroxysteroid sulphotransferase, an enzyme which inactivates many steroids, in particular androgens, in normal breast tissue and in six ductal-type mammary carcinomas using immunohistochemistry. The enzyme is not expressed in the epithelial cells which line the normal breast duct, but is present in significant amounts in neoplastic cells, suggesting that the gene encoding this protein is activated at some stage of the neoplastic transformation. The implications of this finding for the role of steroid metabolism in breast cancer are discussed.

Breast↗

Electroanalytical and biocompatibility studies on carboxylated poly(vinyl chloride) membranes for microfabricated array sensors.

Potassium ion-selective and pH membrane electrodes based on neutral carrier ionophores for K+ (valinomycin) and H+ (TDDA and ETH 5294), respectively, immobilized in carboxylated PVC (PVC-COOH) with normal (classical) and reduced amounts of plasticizer, were investigated with respect to their general analytical performances (linear range, slope, detection limit, selectivity, internal membrane resistance), their biocompatibility and cellular responses. The analytical performance of potassium selective electrodes was not affected by reducing the plasticizer content from 66% (m/m) to about 33% (m/m) while that of pH electrodes was significantly changed at the lower plasticizer concentration level. The adhesive properties of PVC-COOH membranes to an inert substrate such as polyimide-coated Kapton are greatly improved by reducing the plasticizer content of the membrane. In addition, as was reported earlier by this group, improved biocompatibility was observed with these membranes relative to those with increased plasticizer content. A ratio of 1:1 m/m for PVC-COOH to plasticizer is recommended for the construction of planar ISEs without massive use of internal solution.

Animals↗

Characterization of extractable species from poly(etherurethane urea) (PEUU) elastomers.

Methanol extracts of four poly(etherurethane urea) (PEUU) materials were analyzed using Gel Permeation Chromatography (GPC). The additives in the materials were Santowhite powder at 1 wt% and Methacrol 2138 F at 5 wt% loading levels. One-to-two wt% of the original PEUU films was extractable with methanol. The extractables consisted of a low molecular weight (Mw) PEUU polymer, an MDI-rich oligomer, the additives Santowhite (SW) powder and Methacrol 2138 F, and aniline. The low Mw PEUU polymer had a Mw of 12,000 relative to polystyrene, and the MDI-rich oligomer had a Mw of 1000 relative to polystyrene. Quantitation of all extracted species was achieved using GPC; the use of dual-detectors on the GPC made it possible to determine the soft-to-hard composition of the PEUU extracts as a function of molecular weight.

Amines↗

Transesophageal electroventilation. Optimization and improvement.

For electroventilation, short duration pulse trains (0.1 ms) were applied between two axillary pads (transchest) and compared with transesophageal electroventilation where pulses passed between the same pads to an esophageal electrode in apneic, pentobarbitone-anesthetized pigs. Significantly greater tidal volumes were produced by transesophageal electroventilation in comparison with transchest. As measured by maintained tidal volumes, duration of inspiratory air flow, peak flow, percentage hemoglobin oxygen saturation (%SaO2), and end tidal carbon dioxide concentration (ETCO2), the optimal requirements for transesophageal electroventilation were pulse frequency 40 Hz, 0.7 s duration pulses, at 60 to 100 V, pulse width 0.1 ms, with the esophageal electrode proximal to the gastroesophageal junction without producing brachial plexus stimulation. The efficiency of transesophageal electroventilation falls off rapidly following ventricular fibrillation. Thus, the application of this technique would be in respiratory arrest with maintained circulation, eg, in drug-induced respiratory depression, severe smoke inhalation, severe emphysema, high cervical cord lesion, and weaning from prolonged mechanical ventilation.

Animals↗

The changing role of school nurses--one state's experience.

The purpose of this study was to identify the scope of management responsibilities, health education, and early education for handicapped children, as well as clinical and program dimensions of the school nurse role. Changes in responsibilities over the last five years were explored in one state by utilizing a questionnaire aimed at providing a profile of the school nurse's perceived behavioral patterns. In every dimension of the school nurse role included in this study, the responsibilities of the school nurse had either increased or stayed the same on an overall basis.

Adolescent↗

Portable cardiac mapping assessment of acute ischaemic injury.

A portable cardiac mapping system is used to improve the accuracy of diagnosis of acute ischaemic injury outside hospital. Patients presenting chest pain suggestive of myocardial infarction (MI) were mapped by attendant medical personnel operating from a mobile coronary unit. These first MI maps were compared against average normal maps using QRS and ST-T isointegral values. Discriminant function analysis performed on the parameters achieved a sensitivity of 90% and a specificity of 96%.

Body Surface Area↗

Turnover of the photosystem II D1 protein in higher plants under photoinhibitory and nonphotoinhibitory irradiance.

The turnover in vivo of the Photosystem II (PS II) reaction center D1 protein was investigated by [35S] methionine labeling of leaf discs of Brassica napus and subsequent analysis after thylakoid SDS-gel electrophoresis. The rate of D1 protein degradation was found to have a t1/2 of approximately 2 h, at an irradiance corresponding to the growth irradiance. The rate of D1 protein degradation was not increased further by prior photoinhibitory treatment which inactivated 40% of the PS II centers, but the amount of [35S]methionine label incorporated into the D1 protein during 1 h was increased 3-fold. Whether photoinhibited or not, the incorporation of label into the D1 protein increased with irradiance only up to approximately the growth irradiance and above that it decreased with increasing incident light. These data indicate that a high rate of D1 protein degradation occurs not only after photoinhibition but also under conditions where no net decrease in the number of functional PS II centers occurs and that the number of PS II centers that undergo D1 protein turnover varies depending on prior photoinhibitory damage and on incident irradiance. A down-regulation of PS II in leaves at high irradiances is discussed.

Bacterial Proteins↗