Magnetic nanoparticles as gene delivery agents: enhanced transfection in the presence of oscillating magnet arrays.

Magnetic nanoparticle-based gene transfection has been shown to be effective in combination with both viral vectors and with non-viral agents. In these systems, therapeutic or reporter genes are attached to magnetic nanoparticles which are then focused to the target site/cells via high-field/high-gradient magnets. The technique has been shown to be efficient and rapid for in vitro transfection and compares well with cationic lipid-based reagents, producing good overall transfection levels with lower doses and shorter transfection times. In spite of its potential advantages (particularly for in vivo targeting), the overall transfection levels do not generally exceed those of other non-viral agents. In order to improve the overall transfection levels while maintaining the advantages inherent in this technique, we have developed a novel, oscillating magnet array system which adds lateral motion to the particle/gene complex in order to promote transfection. Experimental results indicate that the system significantly enhances overall in vitro transfection levels in human airway epithelial cells compared to both static field techniques (p<0.005) and the cationic lipids (p<0.001) tested. In addition, it has the previously demonstrated advantages of magnetofection-rapid transfection times and requiring lower levels of DNA than cationic lipid-based transfection agents. This method shows potential for non-viral gene delivery both in vitro and in vivo.

Molecular modeling studies of the binding characteristics of phosphates to sevelamer hydrochloride--assessing a novel technique to reduce phosphates contamination.

Recent advances in the use of polymeric materials for the remediation of phosphate from aqueous systems, including biological environments, have prompted the use of computational techniques to determine which methods are feasible to model such complex systems and to model the mechanism of binding action. In particular, Sevelamer Hydrochloride (Renagel) is used as our model polymer. A relatively simple system is constructed with a dimer of Sevelamer Hydrochloride and four phosphate ions used for capture. This work reports on molecular dynamics and Monte Carlo simulations used to determine average structure, and points of intermolecular interaction, and calculate changes in volume after the successful capture of phosphate in our model. Our resulting volume changes are of the order of 20-25%, in comparison to experimental swelling measurements on similar systems, which have an average swelling of 50-60% in aqueous media.

In situ characterization and mapping of iron compounds in Alzheimer's disease tissue.

There is a well-established link between iron overload in the brain and pathology associated with neurodegeneration in a variety of disorders such as Alzheimer's (AD), Parkinson's (PD) and Huntington's (HD) diseases [1]. This association was first discovered in AD by Goodman in 1953 [2], where, in addition to abnormally high concentrations of iron in autopsy brain tissue, iron has also been shown to accumulate at sites of brain pathology such as senile plaques [3]. However, since this discovery, progress in understanding the origin, role and nature of iron compounds associated with neurodegeneration has been slow. Here we report, for the first time, the location and characterisation of iron compounds in human AD brain tissue sections. Iron fluorescence was mapped over a frontal-lobe tissue section from an Alzheimer's patient, and anomalous iron concentrations were identified using synchrotron X-ray absorption techniques at 5 mum spatial resolution. Concentrations of ferritin and magnetite, a magnetic iron oxide potentially indicating disrupted brain-iron metabolism, were evident. These results demonstrate a practical means of correlating iron compounds and disease pathology in-situ and have clear implications for disease pathogenesis and potential therapies.

Detection, identification and mapping of iron anomalies in brain tissue using X-ray absorption spectroscopy.

This work describes a novel method for the detection, identification and mapping of anomalous iron compounds in mammalian brain tissue using X-ray absorption spectroscopy. We have located and identified individual iron anomalies in an avian tissue model associated with ferritin, biogenic magnetite and haemoglobin with a pixel resolution of less than 5 microm. This technique represents a breakthrough in the study of both intra- and extra-cellular iron compounds in brain tissue. The potential for high-resolution iron mapping using microfocused X-ray beams has direct application to investigations of the location and structural form of iron compounds associated with human neurodegenerative disorders--a problem which has vexed researchers for 50 years.

Materials used in urological devices.

The most commonly used materials in the manufacture of urological devices are discussed with respect to their application to urinary catheters, penile implants, testicular implants, artificial urinary sphincters, and urological stents. The manufacturing, physical properties, and general chemical and biological behavior of there materials are reviewed. Attention is paid to future research trends and the general condition of the field of urological device materials.

In vitro measurement of silicone bleed from breast implants.

A method to measure gel bleed from intact silicone gel-filled breast implants was developed. This nondestructive technique permits accurate and reproducible serial measurements of silicone bleed from smooth wall breast implants (n=10) under simulated physiologic conditions in vitro. Gel bleed rates from new low bleed gel-filled implants and intact explants (unbarriered, low bleed, double lumen) were determined. These results demonstrate the reliability of this method to quantify silicone gel bleed and may permit a meaningful comparison of bleed rates from implants in the future.

System for the pH-dependent release of a dye in model dental restorations.

We are developing a system for detecting recurrent caries under dental restorations. The controlled release of dyes under conditions of likely demineralization will alert the dentist to potential secondary caries. Production of acidic species is a characteristic of caries activity; hence, the system uses pH-sensitive polymers to release markers when the pH at the cavity wall of the restored tooth is below 6.5. The objectives of this investigation were to test the hypotheses that (1) the proposed system can be designed to release detectable marker continuously for at least six months in a simulated carious environment, and (2) the transient pH changes in the oral cavity caused by simulated dietary intake will not induce premature marker release from the pH-sensitive polymer placed beneath restorations. Two types of dye-loaded microspheres based on styrene, vinylpyridine, and divinylbenzene were prepared and placed on the floor of model cavity preparations made from an acrylic rod. Each model cavity was restored with a hybrid dental composite, placed in a vial with 5 mL of sodium-lactate/lactic-acid base buffer solution, and stored at 37 degrees C. Solutions of three different pH values were used: 2.86, 4.73, and 6.39. The dye release into storage media was monitored periodically with a UV/VIS spectrophotometer. Results showed that the duration could extend beyond six months for pH > 4.73, and that transient oral pH changes are not likely to result in premature dye release. The data indicate that it would take approximately 21 days for the acidic agent external to the restoration to initiate dye release from restored sites.

Water and abrasive effects on three-body wear of composites.

The degrees of in vitro three-body wear resistance of a hybrid, a small-particle, and a microfilled composite were determined after water storage for up to 24 months. The hybrid composite was the most wear-resistant, while the microfilled composite showed the most wear. The hybrid composite showed no loss of wear resistance as a result of water storage. The small-particle composite showed a decrease in wear resistance after water storage only when tested with silicon carbide abrasive. The wear resistance of the microfilled composite decreased following water storage when tested with either a soft (CaCO3) or a hard (SiC) abrasive. For all composites, the soft abrasive was not capable of causing preferential wear of the polymer matrix, as observed on in vivo specimens. Instead, the filler particles became flattened, with minimal loss of interparticle substance. The hard abrasive did cause preferential wear of the matrix. All composites absorbed water and leached silicon during water storage, indicating that the filler-polymer bond was attacked by hydrolytic degradation. Scanning electron microscopic evaluation of the three-body wear specimens indicated that the in vitro wear method did not duplicate in vivo wear conditions (e.g., the hard abrasive caused excessive wear and chipping of the filler particles in vitro, a pattern that was not usually observed in vivo). Filler-polymer de-bonding was observed on in vivo specimens of all the composites, while it was found only on the in vitro microfilled composite specimens. These findings suggest that filler dislodging is a complex process that cannot be simulated with the in vitro wear method used in this study, not even after prolonged water storage.

Connective tissue repair in circumferential periodontal defects in dogs following use of a biodegradable membrane.

A biodegradable polylactic acid membrane was evaluated for its ability to enhance periodontal regeneration. In 8 Labrador dogs, circumferential defects were created around the maxillary 1st premolars. The defects were combination "1-wall vertical and horizontal". A membrane was adjusted to cover the defect on one side of the jaw, while the contralateral tooth served as control without membrane. After 3 months, healing was evaluated histologically. The results demonstrated that the amount of connective tissue repair with newly formed cementum approximated 50% of the defect height for both test and control groups. Thus, no advantage to the use of membrane was found, which is contradictory to the findings of our previous dog study using the same biodegradable membrane. The possible reasons for this difference in results are discussed relative to suggested mechanisms for membrane effects in periodontal regeneration.

Toxic hydrolysis product from a biodegradable foam implant.

A commercially available mammary implant containing a soft polyester polyurethane foam shell has been examined for possible diamine hydrolysis products. Fourier Transform Infrared (FT-IR) analysis has shown ester and aromatic structures present in the intact foam shell. Vigorous hydrolysis conditions using NaOH have produced a mixture with large amounts of toluene diamine (TDA) present as determined by Gas Chromatography-Mass Spectrometry (GC/MS), and GC/FT-IR. There are numerous clinical reports of the degradation of this implant shell after implantation with incorporation of fragments into phagocytic cells at the implantation site. Recent reports have discussed late pain and other allergic reactions to this implant. Release of the known toxic material, TDA, is consistent with these observations and strongly suggests that an alternative material be used in this application.

The effect of exercise on the circumvaginal muscles in postpartum women.

The effect of exercise on pressure developed by the circumvaginal muscles (CVM) in postpartum women was studied. The CVM assessment system described earlier by Dougherty, Abrams, and McKey used an intravaginal balloon device (IVBD) developed from an impression and model of the vagina. The system (IVBD, pressure transducer and strip chart recorder) provided permanent CVM pressure tracings with high test-re-test reliability (n = 16; r = .85) for maximum pressure (MP). The hypothesis in the research reported here was that exercise with and without an IVBD, when compared to no exercise, would result in significantly higher MP and pressure over time (POT). Forty-five healthy PP volunteers were randomly assigned to a 10-minute per day home training program, for six weeks. The baseline and six-week assessments consisted of CVM pressure tracings during contraction. Dependent variables were MP (highest pressure attained) and POT (area under the pressure curve). Although no significant differences were found between the home training groups, greater improvement was found in the exercise groups. Variability within subjects partly accounts for the results. Pressure changes before and after pregnancy and before and after CVM exercise are demonstrated in a case study. The findings support the use of CVM exercise in the postpartum.

The dynamic characteristics of the circumvaginal muscles.

Circumvaginal muscle (CVM) maximum pressure was studied as related to the postpartal period, parity, type of birth, episiotomy, age, breastfeeding, physical activity, body mass, stress urinary incontinence, and orgasm in a sample of 98 healthy postpartal and nonpostpartal women. Circumvaginal muscle pressure tracings were obtained from these women using a pressure-sensitive, intravaginal balloon device. Correlations between circumvaginal muscle maximum pressures and episiotomy, age, breastfeeding, physical activity, body mass, stress urinary incontinence, and orgasm were either not significant or only weakly significant. However, Duncan's multiple range test showed differences in the maximum pressures between groups (nulliparous, parity greater than 0, cesarean delivery, and vaginal delivery) and indicated that vaginal birth has a marked weakening effect on the circumvaginal muscles.

New attachment formation following controlled tissue regeneration using biodegradable membranes.

The present study evaluated the potential for reestablishment of connective tissue attachment after exclusion of the gingival connective tissue and epithelium during healing after periodontal surgery in a dog model. Buccal mucoperiosteal flaps were elevated in all four premolar regions in two dogs. The buccal bone was reduced to approximately 25% of its original level on 24 roots. Before the elevated flaps were replaced and sutured, membranes were adjusted to cover the exposed root surfaces. On eight surfaces Millipore filters were used and on eight surfaces biodegradable polylactic acid membranes were used. No membranes were placed over the remaining eight root surfaces which served as controls. After two months of healing, the animals were sacrificed and all the roots were subjected to histological analysis. In all of the roots covered with the polylactic acid membrane, newly formed cementum with inserting collagen fibers was observed and covered on the average 46% of the initial defect. Newly formed bone covered 39% of the initial defect. The roots covered by Millipore filters were characterized by significantly less new attachment and bone. In the control roots only minor new attachment was found in the bottom of the defect in some roots. The results suggest that the establishment of a connective tissue attachment is clearly favored by the placement of a polylactic acid membrane to exclude the epithelium and gingival connective tissue during healing.

Custom-made vaginal balloons for strengthening circumvaginal musculature.

A technique for making individualized silicone rubber intravaginal balloons is described. The method entails investing alginate vaginal impressions in silicone rubber and casting a resin model which is then dipped repeatedly in a Silastic Dispersion (Dow Corning, Q7-2213). The range of forces developed by circumvaginal muscles during maximum contractions was determined to be 0.5-4 lbs. Pressure-volume relationships of the balloons showed that the pressure of filling fluid was an accurate replica of the mean pressure in the vagina.

Ultrastructure of whewellite kidney stones: electron-analytical investigation.

The investigation of the degree of interpenetration between the two component phases of whewellite kidney stones, the protein matrix and calcium oxalate monohydrate crystallites, is extended by a technique of microchemical analysis, employing X-ray photoelectron spectroscopy (XPS). The technique involves determining the surface elemental (hence chemical) composition of fractured whewellite stones. Comparison with the bulk composition of about 95% COM and 5% matrix yielded information on the fracture path, hence on the spatial distribution of the two phases. The results showed large increases in matrix signal upon fracture, confirming that the minor phase, matrix, which is expected to provide an easy path for fracture, pervades the structure down to the level of individual crystallites.