Biomedical subjects

M D Evans

Publications and source records attributed to M D Evans.

At least 37 records · Page 2Linked to original sources

PubMed1999

Persistent adhesion of epithelial tissue is sensitive to polymer topography.

The persistent adhesion of corneal epithelial tissue to the surface of a porous polymer is of interest in the development of a corneal onlay. Using an in vitro model system, this study examined the effect of polymer surface topography on the assembly of basement membrane and hemidesmosomes. Corneal epithelial tissue was grown on polycarbonate surfaces with a range of pore sizes (0.1-3.0 micron, pore diameter) and an equivalent nonporous surface. The ultrastructure of the tissue-polymer interface was evaluated using electron microscopy. On the porous surfaces, the tissue responded to a balance between the size of the discontinuity (pores) and the amount of polymer surface between the pores. Continuous basement membrane and a regular pattern of hemidesmosomal plaque occurred only on the 0.1 micron surface, where both the pores and the total surface area covered by pores were relatively small. The assembly of adhesive structures on surfaces with pore diameters between 0.4-2.0 microns was restricted to regions of polymer between pores. No adhesive structures assembled on the nonporous or on the 3.0-micron surface. These results demonstrate that, in addition to porosity, surface topography is a significant factor in the formation of structures involved in the persistent adhesion of stratified epithelial tissue on a polymer.

Animals↗

PubMed1999

Modulation of corneal epithelial stratification by polymer surface topography.

The topography and porosity of a polymer may affect the epithelialization of a corneal implant. We used an in vitro model to examine the effect of polymer surface topography on corneal epithelial tissue stratification and the deposition of proteins associated with epithelial adhesion. A range of topographies was provided by polycarbonate membranes with nominal pore diameters of 0.1, 0.4, 0.8, 1.0, 2.0, or 3.0 microm and a nonporous surface. Stratification of epithelial tissue outgrowth on these surfaces was evaluated using light and electron microscopy. Deposition of proteins associated with basement membrane and adhesion complex formation at the tissue-polymer interface was assessed using immunohistochemistry. Surfaces with pores in the 0.1-0.8-microm-diameter range supported superior stratification and protein deposition compared with those containing pores of > or = 1.0 microm. Cytoplasmic processes penetrated single pores 2.0 and 3.0 microm in diameter and fused pores 1.0 microm in diameter. Tissue on the nonporous surface had a lower level of stratification compared with surfaces with pores 0.1-0.8 microm in diameter. These results point to the significance of surface topography in biomaterial applications that require persistent epithelialization.

Animals↗

PubMed1999

Discrepancies in the measurement of UVC-induced 8-oxo-2'-deoxyguanosine: implications for the analysis of oxidative DNA damage.

Ultraviolet (UV) light-induced indirect, oxidative damage to DNA has received increasing attention with respect to the mutagenic and carcinogenic effects of solar radiation. An oxidative lesion that has raised particular interest because of its qualitative and quantitative importance is 8-oxo-2'-deoxyguanosine. This deoxynucleoside lesion is most frequently measured by high performance liquid chromatography with electrochemical detection (HPLC-EC) following enzymatic hydrolysis of DNA or as the base equivalent, 8-oxoguanine, by gas chromatography-mass spectrometry (GC-MS) following acid hydrolysis of DNA. We have noted a discrepancy in the literature whereby the levels of 8-oxo-2'-deoxyguanosine measured by HPLC-EC in UVC-irradiated DNA are significantly higher than when 8-oxoguanine is measured by GC-MS. By making use of the availability of both HPLC-EC and stable-isotope dilution GC-MS methodologies in our laboratory we have confirmed the discrepancy noted in the literature by parallel analysis of the same UVC-irradiated calf thymus DNA samples. Furthermore, analysis of the UVC-induced product by UV-visible spectrophotometry, voltammetry and its detection by a monoclonal antibody which recognises 8-oxo-2'-deoxyguanosine strongly suggests that the product is indeed 8-oxo-2'-deoxyguanosine. Partial explanation for this discrepancy could be an inordinate resistance of UVC-irradiated DNA to formic acid hydrolysis. However, we cannot completely exclude the possibility that there is a formic acid-labile species which co-elutes with 8-oxo-2'-deoxyguanosine in enzymatically digested UVC-irradiated DNA. Whether this phenomenon is unique to UV-irradiation damage or occurs with other systems that cause oxidative damage to DNA awaits further investigation. Irrespective of the exact mechanism, there will be significant implications for the analysis of oxidative DNA damage.

8-Hydroxy-2'-Deoxyguanosine↗

PubMed1999

Dosimetry of hip irradiation for the prevention of heterotopic bone formation after arthroplasty.

PURPOSE: The dosimetry of hip irradiation for the prevention of heterotopic bone formation following arthroplasty is complicated by the use of custom shielding in the treatment portal, and the fact that irradiation is usually required during a 48 hour period following surgery. Both the machine output and depth dose factors of the resulting fields are modified by the presence of the shielding blocks. A simplified dosimetric approach, based on correction factors for both the output and depth dose as a function of field geometry is presented for various megavoltage energy beams. MATERIALS AND METHODS: Measurements of relative dose factors (RDF) and percentage depth dose (PDD) were carried out for different combinations of field size, block size and separation between adjacent blocks. Both RDF and PDD measurements were made in a water phantom. Ratios of RDF and PDD were obtained by dividing individual measurements or curves by the corresponding values for the open field (i.e., without blocks). The average values of these ratios constitute the correction factors to be applied for a given MU or treatment time calculation. RESULTS: Extensive RDF and PDD measurements reveal that for the field and block dimensions of interest the correction factors for RDF can be parameterized as a function of separation between two adjacent blocks and beam energy alone and the depth correction factors are additionally only a function of depth. The correction factors for depth dose are equally valid for fixed source-skin distance techniques (that use PDD) and fixed source-axis distance techniques (that use TMR). CONCLUSION: A simple model for the calculation of output in hip irradiation is presented for the situation where the use of computer-based algorithms may not be practical. The model accurately predicts the RDF of the treatment portal to within 2% and the PDD to within 2% for the range of field sizes, block sizes, block gaps and beam energies of interest ignoring other variables.

Arthroplasty, Replacement, Hip↗

PubMed1999

Immunochemical detection of glyoxal DNA damage.

The relevance of reactive oxygen species (ROS) in the pathogenesis of inflammatory diseases is widely documented. Immunochemical detection of ROS DNA adducts has been developed, however, recognition of glyoxal-DNA adducts has not previously been described. We have generated a polyclonal antibody that has shown increased antibody binding to ROS-modified DNA in comparison to native DNA. In addition, dose-dependent antibody binding to DNA modified with ascorbate alone was shown, with significant inhibition by desferrioxamine, catalase, and ethanol. Minimal inhibition was observed with uric acid, 1,10-phenanthroline and DMSO. However, antibody binding in the presence of EDTA increased 3500-fold. The involvement of hydrogen peroxide and hydroxyl radical in ascorbate-mediated DNA damage is consistent with ascorbate acting as a reducing agent for DNA-bound metal ions. Glyoxal is known to be formed during oxidation of ascorbate. Glyoxylated DNA, that previously had been proposed as a marker of oxidative damage, was recognised in a dose dependent manner using the antibody. We describe the potential use of our anti-ROS DNA antibody, that detects predominantly Fenton-type mediated damage to DNA and report on its specificity for the recognition of glyoxal-DNA adducts.

Animals↗

PubMed1999

Accuracy of numerically produced compensators.

A feasibility study is performed to assess the utility of a computer numerically controlled (CNC) mill to produce compensating filters for conventional clinical use and for the delivery of intensity-modulated beams. A computer aided machining (CAM) software is used to assist in the design and construction of such filters. Geometric measurements of stepped and wedged surfaces are made to examine the accuracy of surface milling. Molds are milled and filled with molten alloy to produce filters, and both the molds and filters are examined for consistency and accuracy. Results show that the deviation of the filter surfaces from design does not exceed 1.5%. The effective attenuation coefficient is measured for CadFree, a cadmium-free alloy, in a 6 MV photon beam. The effective attenuation coefficients at the depth of maximum dose (1.5 cm) and at 10 cm in solid water phantom are found to be 0.546 cm-1 and 0.522 cm-1, respectively. Further attenuation measurements are made with Cerrobend to assess the variations of the effective attenuation coefficient with field size and source-surface distance. The ability of the CNC mill to accurately produce surfaces is verified with dose profile measurements in a 6 MV photon beam. The test phantom is composed of a 10 degrees polystyrene wedge and a 30 degrees polystyrene wedge, presenting both a sharp discontinuity and sloped surfaces. Dose profiles, measured at the depth of compensation (10 cm) beneath the test phantom and beneath a flat phantom, are compared to those produced by a commercial treatment planning system. Agreement between measured and predicted profiles is within 2%, indicating the viability of the system for filter production.

Feasibility Studies↗

PubMed1999

Validation of a finite element model of the Kirschner-Ehmer external skeletal fixation system.

OBJECTIVE: To determine the validity of finite element analysis (FEA) as a means of examining biomechanical properties of the Kirschner-Ehmer external skeletal fixation system. SAMPLE POPULATION: 10 paired tibiae harvested from skeletally mature dogs weighing between 30 and 38 kg immediately following euthanasia for reasons unrelated to musculoskeletal disease. PROCEDURE: A gap fracture was created in each bone; fragments were stabilized with 3 frame configurations (type I, type II, and type III), using enhanced-profile threaded pins. Each bone-frame construct was tested, using a materials testing machine in 3 modes of testing: axial compression (AC), mediolateral (ML) bending, and craniocaudal (CC) bending, for a total of 9 tests/bone. The elastic limit of the constructs was not exceeded during testing. Mean stiffness values were determined from load-displacement curves. A finite element model of each construct was created, using three-dimensional elastic beam elements, and stiffness values were calculated, using FEA. Correlations between experimental and FEA data then were determined. RESULTS: Significant differences in stiffness were seen among all 3 constructs in CC bending and AC, with stiffness increasing with construct complexity. No significant difference in ML bending stiffness was seen between type-II and type-III constructs; however, both were significantly stiffer than the type-I constructs. The experimental and FEA stiffness data were strongly correlated (AC, r = 0.994; ML bending, r = 0.998; CC bending, r = 0.985). CONCLUSIONS AND CLINICAL RELEVANCE: Strong correlations among experimental and FEA data indicate that FEA is a valid method of comparing stiffness of Kirschner-Ehmer external skeletal fixation constructs.

Animals↗

PubMed1999

Effects of handpiece lubrication on resin-based composite bond strength to enamel.

PURPOSE: To evaluate the effects of handpiece lubrication on the bond strengths of resin-based composite (RBC) to enamel using two different bonding systems. MATERIALS AND METHODS: Sixty extracted noncarious human maxillary incisors were imbedded in acrylic resin tray material with the facial surfaces exposed. 320 grit sandpaper was used to create a flat surface of enamel. The embedded teeth were randomly divided into two bonding systems groups: half used All-Bond 2 and half used OpiBond FL. Within these two groups the teeth were again randomly divided into three subgroups: Dry: 10 teeth prepared with a sterilized unlubricated handpiece; Lube & Run: 10 teeth prepared with a handpiece that was lubricated and run for 30 seconds prior to use; Lube: 10 teeth prepared with a handpiece that was lubricated and not run prior to its use. The enamel was prepared by lightly roughening with a medium grit diamond in the respective handpieces. The preparations were etched for 15 s with 32% phosphoric acid, rinsed with water for 5 s, and dried with compressed air for 5 s. Either bond system was applied and light-cured. A hybrid RBC was placed and light-cured for 60 s. The teeth were stored in water at 37 degrees C 1 week. Shear bond strengths were measured and the collected data subjected to an ANOVA followed by a Tukey's and Bonferroni t-test were performed. RESULTS: The Dry group and the Lube & Run group had stronger mean shear strengths than did the Lube group. The difference between the mean shear strengths for the Dry group and the Lube & Run group was not statistically significant (P < 0.05). A significant difference was found between the Dry group and the Lube group and between the Lube & Run group and the Lube group (P < 0.05). The mean shear strength for All-Bond 2 was higher than for OptiBond FL (P = 0.0054).

Analysis of Variance↗

PubMed1998

Novel repair action of vitamin C upon in vivo oxidative DNA damage.

There appears to be a paucity of data examining the effect of dietary antioxidants on levels of oxidative DNA damage in vivo, limiting evidence-based assessment of antioxidant efficacy, mechanisms and recommendation for optimal intake. We have examined levels of 8-oxo-2'-deoxyguanosine (8-oxodG) in mononuclear cell DNA, serum and urine from subjects undergoing supplementation with 500 mg/day vitamin C. Significant decreases in DNA levels of 8-oxodG were seen, correlating strongly with increases in plasma vitamin C concentration. Furthermore we established a timecourse for sequential, significant increases in serum and urinary 8-oxodG levels. These results illustrate, for the first time in humans, the kinetics of 8-oxodG removal and processing in vivo, suggesting a role for vitamin C in the regulation of DNA repair enzymes and thereby demonstrating a non-scavenging antioxidant effect.

8-Hydroxy-2'-Deoxyguanosine↗

PubMed1998

Polymer surface chemistry and a novel attachment mechanism in corneal epithelial cells.

The initial attachment reaction of most cultured cell types to polymers is based on a linkage of integrin receptors to serum-derived fibronectin and vitronectin that adsorb onto the polymer surface. Recently isolated corneal epithelial cells have an additional attachment mechanism, known to operate on tissue culture polystyrene, which involves endogenous protein synthesis and an intact system of microtubules. Here, we determine if this novel attachment mechanism is operative on polymers of different surface chemistries. The attachment, growth, and deposition of basement membrane proteins by corneal epithelial cells was compared on two hydrophilic surfaces (tissue culture polystyrene and Primaria) and one relatively hydrophobic surface (unmodified polystyrene). Superior levels of cell attachment were found on the hydrophilic polymers, but cells also attached effectively to the hydrophobic surface. Growth rates showed that the cells were able to overcome the differential effects of polymer surface chemistry during a 7-day time period. Polymer surface chemistry had subtle effects on the temporal pattern of biosynthesis of extracellular matrix proteins likely to be involved in cell adhesion. These results show that effective attachment and growth can occur on a hydrophobic polymer when corneal epithelial cells use the endogenous attachment mechanism.

Animals↗

PubMed1998

Persistent adhesion of corneal epithelial tissue on synthetic lenticules in vivo.

PURPOSE: In intact cornea, firm anchorage of the epithelium to its underlying stroma is provided by a system of adhesive structures, which include the epithelial basement membrane, adhesion complexes and associated alpha6beta4 integrin receptors. In the present study we sought to determine whether the sustained epithelialization of synthetic lenticules observed in vivo involved the formation of adhesive structures at the epithelial-lenticule interface similar to those present in intact tissue. METHODS: Collagen I-coated microporous polycarbonate membranes were implanted in the anterior stroma of adult cats. Successfully epithelialized lenticules were maintained on eye for 9-12 weeks, after which time the epithelial-lenticule interface was examined for evidence of adhesive structures using immunohistochemistry and electron microscopy. RESULTS: Immunohistochemistry identified laminin, hemidesmosomal plaque, collagen VII and alpha6 integrin at the tissue lenticule interface. Ultrastructural examination showed evidence of assembly of these components into a basement membrane and adhesion complexes. CONCLUSIONS: The formation of these adhesive structures is likely to have contributed to the sustained epithelialization observed clinically on the collagen-coated synthetic lenticules.

Animals↗

PubMed1998

Verification of the correspondence between CT-stimulated and treatment beams.

A single phantom technique has been developed to verify the full CT simulation and treatment-beam delivery procedures. The phantom consists of a target delineated by thin copper strips, affixed to therapy verification film, and inserted securely between two slices of water-equivalent material. The target is defined with the aid of the copper strips, and the position of the isocenter and beam parameters such as field size, and gantry and collimator angle are determined by CT simulation. With these parameters, the phantom is subsequently irradiated by the linear accelerator in the treatment position. Correspondence between the planned and the irradiated region is determined by the position of the copper strips on the film. The technique is a simple and practical method for verifying the entire CT simulation and treatment-beam delivery processes, and provides a permanent record of the correspondence between the planning digitally reconstructed radiographs (DRR) and the actual beam delivered.

Computer Simulation↗

PubMed1998

Superficial and orthovoltage x-ray beam dosimetry.

Output of superficial and orthovoltage x-ray units may be measured with cylindrical or end-window parallel-plate ionization chambers. The air-kerma calibration factors for these chambers are usually determined free in air, and the x-ray machine output is stated as the air-kerma rate free in air, which, when multiplied with the appropriate backscatter factor, gives the air-kerma rate on the surface of a phantom or patient. For end-window chambers, especially when they are used for measurements of small fields or low x-ray energies, the air-kerma calibration factors may also be determined with the chamber embedded in a tissue-equivalent phantom. This results in field size dependent air-kerma in-air calibration factors but obviates the requirement for knowledge of back-scatter factors when determining the air-kerma rate on the surface of a phantom. Since there still is considerable uncertainty in tabulated backscatter factors as a function of field size and x-ray beam energy, the output measurement technique which determines the air-kerma rate on phantom surface with a phantom-embedded end-window ionization chamber offers a clear advantage over the in-air calibration method.

Air↗

PubMed1997

Surface applicators for high dose rate brachytherapy in AIDS-related Kaposi's sarcoma.

PURPOSE: The development of commercially available surface applicators using high dose rate remote afterloading devices has enabled radiotherapy centers to treat selected superficial lesions using a remote afterloading brachytherapy unit. The dosimetric parameters of these applicators, the clinical implementation of this technique, and a review of the initial patient treatment regimes are presented. METHODS AND MATERIALS: A set of six fixed-diameter (1, 2, and 3 cm), tungsten/steel surface applicators is available for use with a single stepping-source (Ir-192, 370 GBq) high dose rate afterloader. The source can be positioned either in a parallel or perpendicular orientation to the treatment plane at the center of a conical aperture that sits at an SSD of approximately 15 mm and is used with a 1-mm thick removable plastic cap. The surface dose rates, percent depth dose, and off-axis ratios were measured. A custom-built, ceiling-mounted immobilization device secures the applicator on the surface of the patient's lesion during treatment. RESULTS: Between November 1994, and September 1996, 16 AIDS-related Kaposi's sarcoma patients having a total of 120 lesions have been treated with palliative intent. Treatment sites were distributed between the head and neck, extremity, and torso. Doses ranged from 8 to 20 Gy, with a median dose of 10 Gy delivered in a single fraction. Treatments were well tolerated with minimal skin reaction, except for patients with lesions treated to 20 Gy who developed moderate/severe desquamation. CONCLUSION: Radiotherapy centers equipped with a high dose rate remote afterloading unit may treat small selected surface lesions with commercially available surface applicators. These surface applicators must be used with a protective cap to eliminate electron contamination. The optimal surface dose appears to be either 10 or 15 Gy depending upon the height of the lesion.

Acquired Immunodeficiency Syndrome↗

PubMed1997

The prediction of late rectal complications in patients treated with high dose-rate brachytherapy for carcinoma of the cervix.

PURPOSE: The aim of this work is to investigate an unusually high rate of late rectal complications in a group of 43 patients treated with concomitant irradiation and chemotherapy for carcinoma of the cervix between December 1988 and April 1991, with a view to identifying predictive factors. METHODS AND MATERIALS: The biologically effective dose received by each patient to the rectal reference point defined by the International Commission of Radiation Units and Measurements, Report 38, were calculated. Radiotherapy consisted of 46 Gy external beam irradiation plus three high dose-rate intracavitary treatments of 10 Gy each prescribed to point A. Cisplatin 30 mg/m2 was given weekly throughout the duration of the irradiation. The results have been compared to data from 119 patients treated with irradiation alone to assess the confounding effect of the cisplatin. RESULTS: The relationship between the biologically effective dose delivered to the rectal reference point and the development of late complications shows a strong dose-response with a threshold for complications occurring at approximately 125 Gy3 corresponding to a brachytherapy dose of approximately 8 Gy per fraction. This value is approximately the same biologically effective dose threshold as that found for external beam irradiation in the head and neck region. The data from the group of patients treated without cisplatin is comparable to the data from the first group of patients in the lower dose ranges; the higher doses were not used and thus are not available for comparison. CONCLUSION: Using the linear quadratic model applied to our clinical results, we have established a threshold for late rectal complications for patients treated with external beam irradiation and high dose-rate brachytherapy for carcinoma of the cervix. This threshold is consistent with similar data for external beam irradiation in the head and neck region.

Antineoplastic Agents↗

PubMed1997

Multiple attachment mechanisms of corneal epithelial cells to a polymer--cells can attach in the absence of exogenous adhesion proteins through a mechanism that requires microtubules.

The initial adhesion of epithelial cells is recognized as a critical determinant in the epithelialization of a polymer. Previously, the attachment of a variety of cell types to a polymer has been shown to be mediated by fibronectin and/or vitronectin that adsorb onto the polymer surface from the serum used to supplement the culture medium. Such an attachment reaction, dependent upon exogenous cell-adhesive proteins, is likely to involve the actin cytoskeleton and integrin receptors on the cell surface. In the current study, we have examined the attachment of recently isolated corneal epithelial cells to tissue culture polystyrene in the absence of such exogenous cell-adhesive proteins. Under these circumstances, there is an alternative mechanism of attachment operative between corneal epithelial cells and the polymer surface which is essentially independent of the adsorption of serum-derived fibronectin and vitronectin. This is an unusual phenomenon not previously observed with other normal cell types. The presence of cycloheximide inhibited this alternative attachment mechanism in corneal epithelial cells, indicating the role of newly synthesized proteins in this reaction. Additionally, we found that cell attachment to the polymer surface in the absence of cell-adhesive proteins was substantially inhibited by the presence of demecolcine, but not by cytochalasin B, thereby demonstrating the involvement of microtubules rather than actin microfilaments in this reaction. In the presence of serum-derived fibronectin and vitronectin, the attachment of corneal epithelial cells to a polymer may involve dual attachment mechanisms.

Actins↗

PubMed1997

Systematic and random error in repeated measurements of temporal and distance parameters of gait after stroke.

OBJECTIVE: To obtain intersession estimates of error for temporal and distance (TD) parameters of gait in a sample of stroke patients undertaking inpatient rehabilitation. DESIGN: Thirty-one stroke patients were measured with an instrumented footswitch system (after a median of 46 days poststroke; interquartile range = 26 to 63) walking over a 10-meter distance a total of four times on 3 consecutive days. Two familiarization walks provided intrasession retest data. RESULTS: Metric estimates of systematic and random error have been provided for obtained TD parameters. Proportional indices of reliability (ICC [2,1] and Pearson's r) were generally high, ranging from .72 to .94. CONCLUSION: By quantifying systematic and random error associated with the process of repeated measurements, criteria have been provided for evaluating change in TD variables during rehabilitation. Although error for gait velocity was small relative to individual differences in the stroke group, it was large relative to levels of change derived from measurements reported during typical periods of rehabilitation. Serial measurements of gait during rehabilitation may be better than two consecutive measurements. This study highlights the need to interpret estimates of error according to the purpose of measurement.

Aged↗

PubMed1997

Analysis of urinary pseudouridine by micellar electrokinetic capillary chromatography.

We describe a capillary electrophoresis procedure, using uridine as an internal standard, for the analysis of urinary pseudouridine following solid-phase extraction. This method retains the advantages of existing chromatographic techniques but has superior resolving power and is technically less demanding. The standard curves were linear and reproducible with a detection limit of 60 fmol; chromatographic analysis was complete in under 10 min. Injection variability was < 5% and multiple independent analyses of the same urine sample for pseudouridine concentration gave coefficients of variation of < 10%. The mean (SD) urinary pseudouridine level in 18 healthy subjects was 16.1 (2.1) nmol/mumol creatinine. For a limited group of subjects where samples were taken more frequently, intra-individual variation averaged 27.5% reflecting variable excretion.

Adolescent↗