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A prospective randomised clinical trial of one bis-GMA-based and two ormocer-based composite restorative systems in class II cavities: three-year results.

OBJECTIVES: Ormocer composites, consisting of a silicon-based polymer, have been developed recently as a tooth-coloured restorative material. The purpose of this prospective randomised clinical trial was to evaluate the performance of two small-particle hybrid ormocer-based restorative systems (AD, Admira/Admira Bond, VOCO; DE, Definite/Etch & Prime 3.0, Dentsply) and one small-particle hybrid bis-GMA-based composite restorative system (TC, Tetric-Ceram/Syntac, Ivoclar-Vivadent) in occlusal stress-bearing restorations. METHODS: One hundred and twenty-eight occlusal-proximal restorations (44 AD, 43 DE and 41 TC) were placed according to the manufacturer's instructions in thirty-two adult patients. Their clinical performance was scored according to the USPHS criteria and evaluation of bite-wing radiographs. RESULTS: After 3 years, four AD, five DE and four TC restorations had failed due to fracture or marginal gap formation. Surface roughness improved significantly when compared to the baseline in AD and TC (Friedman test, p<0.05) during the first year but returned to baseline values after 3 years. DE had a significant tendency towards discolouration (p<0.05). Bite-wing radiographs showed two AD and one TC restorations with internal porosities. ANOVA showed that larger restorations (> or = 3 surfaces) showed significantly more degradation than smaller ones. CONCLUSIONS: In a group of class II restorations, there was no significant difference in failures after 3 years between ormocer-based and bis-GMA-based restorative systems.

Acrylates↗

A study on grafting and characterization of HMDI-modified calcium hydrogenphosphate.

It is known that the organic molecules can provide an effective means to manipulate the surface properties of the biodegradable ceramic. There are two ways to modify the surface of the biodegradable ceramic by organic molecules. The first one is through surface adsorption but organic molecules will easily be washed out in the physiological environment. The second approach is to graft organic molecules through covalent bond to the hydroxyl groups that are available on the surface of the ceramics. Isocyanate group has been reported as a coupling agent for hydroxyapatite and organic molecule. The studies showed that the isocyanate could react with hydroxyl groups of hydroxyapatite and form a covalent bond between isocyanate and hydroxyapatite. In the study, hexamethylene diisocyanate (HMDI) was used as coupling agent and calcium hydrogenphosphate (CaHPO4, CHP) was the candidate ceramic. CHP will react with HMDI at the temperature of 20 degrees C, 30 degrees C, 40 degrees C, 50 degrees C, 60 degrees C, and 70 degrees C for 4h. Dibutyltin dilaurate and hydroquinone were used as catalyst and inhibitor, respectively. The effect of reaction temperature on the grafted yield will be described. The linkage between CHP and HMDI will be characterized by DTA, TGA, FTIR, XRD, and 31P, 13C liquid state NMR. From the results, we successfully modified the surface of CHP with coupling agent of HMDI. The grafted yield of HMDI on CHP was increasing with the reaction temperature. The best temperature for CHP modified by HMDI is around 50 degrees C. The linkage between HMDI and the surface of CHP is a urethane linkage as CHP-O-CO-NH-(CH2)6-N=C=O. After further treatment, the terminal group of CHP treated with HMDI (MCHP) will be converted into a primary amine group as the formula of CHP-O-CO-NH-(CH2)6-NH2. If reaction temperature is 60 degrees C, long extension chain will occur with a urea linkage between the isocyanate groups as the formula of CHP-O-CO-NH-(CH2)6-(NH-CO-NH-(CH2)6)n-NH2. At reaction temperature higher than 60 degrees C, the HMDI will become prepolymerized forms in solution. The prepolymerized forms such as allophanate, biuret, uretidione and urea linkage will turn the solution into gel type mixture, which will lead to low grafted yield of HMDI on CHP. When MCHP prepared at the temperature 20 degrees C, there is no evidence of long extension but the grafted yield is the lowest only 0.9 wt% around.

Calcium Phosphates↗

[Degradation of nitrobenzene in aqueous solution by modified ceramic honeycomb-catalyzed ozonation].

Comparative experiments of modified ceramic honeycomb, ceramic honeycomb-catalyzed ozonation and ozonation alone were conducted with nitrobenzene as the model organic pollutant. It was found that the processes of modified ceramic honeycomb and ceramic honeycomb-catalyzed ozonation could increase the removal efficiency of nitrobenzene by 38.35% and 15.46%, respectively, compared with that achieved by ozonation alone. Under the conditions of this experiment, the degradation rate of modified ceramic honeycomb-catalyzed ozonation increased by 30.55% with the increase of amount of catalyst to 5 blocks. The degradation rate of three process all increased greatly with the increase of temperature and value of pH in the solution. But when raising the pH value of the solution to 10.00, the advantage of modified ceramic honeycomb-catalyzed ozonation processes lost. The experimental results indicate that in modified ceramic honeycomb-catalyzed ozonation, nitrobenzene is primarily oxidized by *OH free radical in aqueous solution. The adsorption of nitrobenzene is too limited to have any influence on the degradation efficiency of nitrobenzene. With the same total dosage of applied ozone, the multiple steps addition of ozone showed a much higher removal efficiency than that obtained by one step in three processes. Modified ceramic honeycomb had a relative longer lifetime.

Catalysis↗

Better chemistry through ceramics: the physical bases of the outstanding chemistry of ORMOSIL.

Twenty years after their invention, sol-gel organically modified silicates (ORMOSIL) are finding a number of impressive applications that range from efficient deliver of genes into mouse brains to self-ordered helices of interest to fields as diverse as optics, catalysis, molecular recognition, and chromatography. The physical bases of this mulifaceted chemistry, therefore, are of immense importance to scientists working toward new applications such as photovoltaics and catalysis that are crucially important in making sustainable global development. The purpose of this article is to provide a general picture of ORMOSIL's physical chemistry that will be useful in the creative development of new materials capable to solve a number of relevant open problems.

Journal Article↗

Effects of dental materials on insulin release from isolated islets of Langerhans.

OBJECTIVES: To investigate the possibility of using a whole organ model for evaluating the biological effects of dental restoration materials in vitro. METHODS: The effect on insulin release of isolated Langerhans islets of a series of dental materials was examined. The islets were incubated for 1h with extracts obtained from various dental materials and insulin was assayed radioimmunologically with crystalline mouse insulin. The results were analysed statistically using analysis of variance (ANOVA) and Scheffe's test at a significance level of p < 0.05. RESULTS: One dental ceramic, Vita VMK 95, significantly (p < 0.01) decreased the insulin release, whereas another dental ceramic, Empress (p < 0.01), a partly re-cast high-noble gold alloy (p < 0.001), a modified high-noble gold alloy (p < 0,05), and unalloyed copper (p < 0.001) significantly increased the release of insulin. SIGNIFICANCE: The results demonstrate a new instance of examining the biological effects of dental restoration materials. The method provides information about the effect of different materials on organ level in vitro that can complement other in vitro tests.

Aluminum Oxide↗

Analysis of non-covalent protein complexes by capillary electrophoresis--time-of-flight mass spectrometry.

A capillary electrophoresis-electrospray ionisation time-of-flight mass spectrometry (CE-ESI-TOF-MS) method for characterisation of non-covalent protein complexes is described using a coaxial liquid sheath-flow sprayer. The CE capillary was connected to the mass spectrometer using a commercial CE-MS sprayer mounted on a ceramic holder of the ESI interface of the mass spectrometer. Using myoglobin (Mb) as an example of non-covalent protein complex, the effect on complex stability caused by organic modifiers added to the sheath liquid was analysed. Depending on the amount of methanol, either intact Mb or the apoprotein and the prosthetic heme group were detected.

Electrophoresis, Capillary↗

An investigation into the effect of a ceramic particle trap on the chemical mutagens in diesel exhaust.

Diesel exhaust particles and vapor phase samples were collected from the diluted (15:1) exhaust of a 10.4 L displacement medium-duty engine (Caterpillar 3208), operated under EPA steady-state cycle Modes 4 and 5 conditions for load (50 and 75 percent, respectively) and speed (1680 rpm). Baseline (uncontrolled) emissions were compared to the exhaust modified by the use of an uncatalyzed monolithic ceramic trap (Corning). The Salmonella/microsome mutagenicity bioassay (Ames Test) was used to direct the course of chemical analyses. Total particulate matter (TPM), soluble organic fraction (SOF) (from TPM), sulfate fraction (SO4) (from TPM), and solid fraction (SOL) (from particle) were determined from dilute exhaust particles collected on 47 mm Teflon-coated woven glass fiber filters. Coincidentally, particles were collected on 508 x 508 mm Teflon-coated non-woven glass fiber filters, and vapor-phase samples were collected on XAD-2 resin. The SOF and VOC for chemical and biological characterization were obtained by Soxhlet extraction of samples with dichloromethane (DCM). Hydrocarbon mass balances were developed to evaluate the efficiency of the sampling system. Use of the ceramic traps caused no change in engine total hydrocarbon (HC) levels at Mode 4 but decreases in TPM, SOF, and NO2 were noted. In terms of HC emissions only, the percentage of SOF was significantly reduced, but the percentage of VOC was unchanged. For Mode 5, the engine HC levels were significantly reduced but the proportions of HC components, i.e. the percentage of SOF and the percentage of VOC, did not change significantly. Engine emission levels of TPM, SOF, and nitrogen dioxide (NO2) were also significantly reduced at Mode 5. At both Modes 4 and 5, use of the ceramic particle traps caused an increase in the direct-acting (TA98) mutagenicity (revertants/microgram) of the SOF and a decrease in the activity of the VOC. The traps caused a 70 percent reduction of TPM at Mode 4 but only a 45 percent reduction in particulate-associated direct-acting mutagenicity on the basis of raw exhaust emissions (kRevertants/m3). At Mode 5 with the traps, there was an 85 percent reduction in TPM and only a 25 percent reduction in the activity of the SOF. The direct-acting mutagenicity of the VOC was reduced by use of the traps by 40 and 65 percent (kRevertants/m3) for Modes 4 and 5, respectively. In contrast, the indirect-acting mutagenicity of the Mode 4 VOC increased nearly 150 percent. Filter loading and reexposure experiments indicated that sampling artifacts did not contribute to the SOF mutagenicity at Mode 4.(ABSTRACT TRUNCATED AT 400 WORDS)

Ceramics↗

Amperometric hydrogen peroxide sensor based on a sol-gel-derived ceramic carbon composite electrode with toluidine blue covalently immobilized using 3-aminopropyltrimethoxysilane.

A carbon composite amperometric hydrogen peroxide sensor has been developed using a sol-gel technique. Toluidine blue (TB), which acts as the redox mediator, was covalently immobilized via glutaraldehyde crosslinking with an organically modified silane, namely 3-aminopropyltrimethoxysilane (APTMOS). Methyltrimethoxysilane (MTMOS) was used as the additional monomer; this controls the hydrophobicity of the electrode surface, thus limiting the wettability. The immobilization of TB within the sol-gel matrix was confirmed with FTIR studies. The sol-gel mixture containing TB immobilized in APTMOS and MTMOS was mixed with graphite powder in order to prepare the carbon composite electrode. The electrode was characterized using voltammetric techniques and its electrocatalytic activity for the reduction of hydrogen peroxide was also studied. The carbon composite electrode has the advantage of sensing H(2)O(2) at a lower potential and with a higher sensitivity, and interferences due to ascorbic acid, uric acid and acetaminophen were greatly minimized. The linear range for the determination of H(2)O(2) extends from 5.37 x 10(-6) to 6.15 x 10(-3) M, with a correlation coefficient of 0.9981. The detection limit was found to be 2.15 x 10(-6) M. The covalent immobilization of TB effectively prevents the leakage of the water-soluble mediator during measurements. The modified electrode, aside from electrocatalyzing the reduction of H(2)O(2), exhibits distinct advantages in terms of surface renewal in the event of surface fouling, as well as simple preparation, good chemical and mechanical stability, and good reproducibility.

Biosensing Techniques↗

Macroporous hydroxyapatite as alloplastic material for dental applications.

In the dentistry field, synthetic hydroxyapatite can be conveniently used as semiabsorbable alloplastic material to solve a number of clinical problems. The hydroxyapatite structure influences bone ingrowth as well as its resorption. In particular, pore size must exceed 100 microm to allow new bone ingrowth. Because of its brittleness, highly porous hydroxyapatite is difficult to handle without causing damage, and, therefore, its mechanical performance has to be improved placing the material on a dense hydroxyapatite substrate. Dense/porous hydroxyapatite laminates can be obtained by slip casting technology. The method starts with the preparation of a ceramic powder-binder-solvent system. This slurry produces, after solvent evaporation, a soft green tape, that is successively baked at 300 degrees C to remove all organic component, and finally sintered at high temperature (1200 degrees C). The material porosity cannot be significantly modified by changes in the slurry composition and sintering temperature; therefore, the macroporous hydroxyapatite layer must be obtained using a porous hydroxyapatite powder. The powder is prepared by grinding of green tape pieces and the resulting coarse material is successively baked at 300 degrees C. Such material is mixed with a polymer solution and cast on a green tape substrate. Layer fractures are not observed in both film bulks and interface, because during sintering the consolidation of two layers happens simultaneously. In the resulting material, the first layer consisted of macroporous hydroxyapatite with high osteoconductive properties, and the second layer was a dense hydroxyapatite substrate able to improve the laminate mechanical properties.

Journal Article↗

A bioactive titanium foam scaffold for bone repair.

While titanium has been clinically successful as an orthopedic or dental implant material, performance problems still persist related to implant-bone interfacial strength and mechanical modulus mismatch between titanium and tissue. We describe here the preparation of a titanium foam as a better mechanical match to tissue with surfaces attractive to bone cells through deposition of an organically-modified apatite layer (organoapatite). In a rotating bioreactor, these organoapatite-coated foams are successfully colonized by preosteoblastic cells. Finite element analyses suggest that ingrown tissue in these systems may improve both implant performance and tissue formation through load-sharing and stress distribution. The novel metal-ceramic-polymer hybrid materials described here hold great promise for bone tissue engineering.

3T3 Cells↗

Biological reactivity of zirconia-hydroxyapatite composites.

Materials and devices intended for end-use applications as implants and medical devices must be evaluated to determine their biocompatibility potential in contact with physiological systems. The use of standard practices of biological testing provides a reasonable level of confidence concerning the response of a living organism to a given material or device, as well as guidance in selecting the proper procedures to be carried out for the screening of new or modified materials. This article presents results from cytotoxicity assays of cell culture, skin irritation, and acute toxicity by systemic and intracutaneous injections for powders, ceramic bodies, and extract liquids of hydroxyapatite (HA), calcia partially stabilized zirconia (ZO), and two types of zirconia-hydroxyapatite composites (Z4H6 and Z6H4) with potential for future use as orthopedic and dental implants. They indicate that these materials present potential for this type of application because they meet the requirements of the standard practices recommended for evaluating the biological reactivity of ATCC cell cultures (CCL1 NCTC clone 929 of mouse connective tissue and CCL 81 of monkey connective tissue) and animals (rabbit and mouse) with direct or indirect patient contact, or by the injection of specific extracts prepared from the material under test. In addition, studies involving short-term intramuscular and long-term implantation assays to estimate the reaction of living tissue to the composites studied, and investigations on long-term effects that these materials can cause on the cellular metabolism, are already in progress.

Animals↗

Nanostructured hybrid materials from aqueous polymer dispersions.

Organic-inorganic (O-I) hybrids with well-defined morphology and structure controlled at the nanometric scale represent a very interesting class of materials both for their use as biomimetic composites and because of their potential use in a wide range of technologically advanced as well as more conventional application fields. Their unique features can be exploited or their role envisaged as components of electronic and optoelectronic devices, in controlled release and bioencapsulation, as active substrates for chromatographic separation and catalysis, as nanofillers for composite films in packaging and coating, in nanowriting and nanolithography, etc. A synergistic combination or totally new properties with respect to the two components of the hybrid can arise from nanostructuration, achieved by surface modification of nanostructures, self-assembling or simply heterophase dispersion. In fact, owing to the extremely large total surface area associated with the resulting morphologies, the interfacial interactions can deeply modify the bulk properties of each component. A wide range of starting materials and of production processes have been studied in recent years for the controlled synthesis and characterization of hybrid nanostructures, from nanoparticle or lamellar dispersions to mesoporous materials obtained from templating nanoparticle dispersions in a continuous, e.g. ceramic precursor, matrix. This review is aimed at giving some basic definitions of what is intended as a hybrid (O-I) material and what are the main synthetic routes available. The various methods for preparing hybrid nanostructures and, among them, inorganic-organic or O-I core-shell nanoparticles, are critically analyzed and classified based on the reaction medium (aqueous, non-aqueous), and on the role it plays in directing the final morphology. Particular attention is devoted to aqueous systems and water-borne dispersions which, in addition to being environmentally more acceptable or even a mandatory choice for any future development of large output applications (e.g. in paint, ink and coating industry), can provide the thermodynamic drive for self-assembling of amphiphilics, adsorption onto colloidal particles or partitioning of the hybrid's precursors between dispersed nanosized reaction loci, as in emulsion or miniemulsion free-radical polymerization. While nanoencapsulation and self-assembling processes are already exploited as commercially viable fabrication methods, a newly developed technique based on two-stage sol-gel and free-radical emulsion polymerization is described, which can grant a versatile synthetic approach to hybrid O-I nanoparticles with tailor-made composition of both the organic core and the silica or organosilica shell, and good control on morphology, size and heterophase structure in the 50-500 nm range. Styrene or acrylate homo- and copolymer core latex particles need to be modified with a reactive comonomer, such as trimethoxysilylpropyl methacrylate, to achieve efficient interfacial coupling with the inorganic shell. Accurate control over pH and process conditions is required to avoid latex coagulation or, in case of organic particles with uniform composition, incipient intraparticle crosslinking.

Journal Article↗

Healing of critically sized femoral defects, using genetically modified mesenchymal stem cells from human adipose tissue.

The FDA has approved the clinical use of recombinant bone morphogenetic proteins (BMPs). However, the use of recombinant BMPs in humans has required large doses of the proteins to be effective, which suggests that the delivery method of bone morphogenetic proteins needs to be optimized. Gene therapy is an alternative method to deliver such recombinant proteins, and gene transfer techniques have been tested on a variety of cell types including bone marrow cells, skin fibroblasts, peripheral blood monocytes, and muscle-derived cells. In this study, we sought to determine the ability of BMP-2-producing human adipose-derived mesenchymal stem cells to heal a critically sized femoral defect in a nude rat model. After approval by the human subjects protection committee, human adipose tissue was obtained from healthy donors. The lipoaspirate was processed as previously described (De Ugarte, D.A., et al. Cells Tissues Organs 174, 101, 2003). Cells were grown in culture and infected with a BMP-2-carrying adenovirus. Five million cells were applied to a collagen- ceramic carrier and implanted into femoral defects as previously described (Zuk, P.A., et al. Mol. Biol. 13, 4279, 2002). All animals were killed at 8 weeks. Femora were dissected out and underwent radiographic, histologic, and biomechanical analysis. Eleven of the 12 femora in the group treated with human processed lipoaspirate (HPLA) cells genetically modified to overexpress BMP-2 had healed at 8 weeks. This was assessed by radiographs, by mechanical testing, and by histology. The one femur that did not heal had a subacute infection. All eight of the femora treated with the rhBMP-2-impregnated collagen-ceramic carrier healed. No statistically significant difference was detected between these two groups. Evaluation of the control groups: group II (collagen- ceramic carrier with HPLA cells) and group III (collagen-ceramic carrier alone) showed that none of the femora had healed by 8 weeks. Our results indicate that HPLA cells genetically modified by adenoviral gene transfer to overexpress BMP-2 can induce bone formation in vivo and heal a critically sized femoral defect in an athymic rat. The HPLA cells alone did not induce significant bone formation. However, when combined with an osteoinductive factor these cells may be an effective method for enhancing bone healing and the tissue engineering of bone.

Adenoviridae↗

Ceramic-based nanoparticles entrapping water-insoluble photosensitizing anticancer drugs: a novel drug-carrier system for photodynamic therapy.

A novel nanoparticle-based drug carrier for photodynamic therapy is reported which can provide stable aqueous dispersion of hydrophobic photosensitizers, yet preserve the key step of photogeneration of singlet oxygen, necessary for photodynamic action. A multidisciplinary approach is utilized which involves (i) nanochemistry in micellar cavity to produce these carriers, (ii) spectroscopy to confirm singlet oxygen production, and (iii) in vitro studies using tumor cells to investigate drug-carrier uptake and destruction of cancer cells by photodynamic action. Ultrafine organically modified silica-based nanoparticles (diameter approximately 30 nm), entrapping water-insoluble photosensitizing anticancer drug 2-devinyl-2-(1-hexyloxyethyl) pyropheophorbide, have been synthesized in the nonpolar core of micelles by hydrolysis of triethoxyvinylsilane. The resulting drug-doped nanoparticles are spherical, highly monodispersed, and stable in aqueous system. The entrapped drug is more fluorescent in aqueous medium than the free drug, permitting use of fluorescence bioimaging studies. Irradiation of the photosensitizing drug entrapped in nanoparticles with light of suitable wavelength results in efficient generation of singlet oxygen, which is made possible by the inherent porosity of the nanoparticles. In vitro studies have demonstrated the active uptake of drug-doped nanoparticles into the cytosol of tumor cells. Significant damage to such impregnated tumor cells was observed upon irradiation with light of wavelength 650 nm. Thus, the potential of using ceramic-based nanoparticles as drug carriers for photodynamic therapy has been demonstrated.

Antineoplastic Agents↗

Harvesting energy from the marine sediment-water interface II. Kinetic activity of anode materials.

Here, we report a comparative study on the kinetic activity of various anodes of a recently described microbial fuel cell consisting of an anode imbedded in marine sediment and a cathode in overlying seawater. Using plain graphite anodes, it was demonstrated that a significant portion of the anodic current results from oxidation of sediment organic matter catalyzed by microorganisms colonizing the anode and capable of directly reducing the anode without added exogenous electron-transfer mediators. Here, graphite anodes incorporating microbial oxidants are evaluated in the laboratory relative to plain graphite with the goal of increasing power density by increasing current density. Anodes evaluated include graphite modified by adsorption of anthraquinone-1,6-disulfonic acid (AQDS) or 1,4-naphthoquinone (NQ), a graphite-ceramic composite containing Mn2+ and Ni2+, and graphite modified with a graphite paste containing Fe3O4 or Fe3O4 and Ni2+. It was found that these anodes possess between 1.5- and 2.2-fold greater kinetic activity than plain graphite. Fuel cells were deployed in a coastal site near Tuckerton, NJ (USA) that utilized two of these anodes. These fuel cells generated ca. 5-fold greater current density than a previously characterized fuel cell equipped with a plain graphite anode, and operated at the same site.

Electrochemistry↗

Synthesis of bioactive organic-inorganic nanohybrid for bone repair through sol-gel processing.

So-called bioactive ceramics have been attractive because they form bone-like apatite on their surfaces to bond directly to living bone when implanted in bony defects. However, they are much more brittle and much less flexible than natural bone. Organic-inorganic hybrids consisting of flexible organic polymers and the essential constituents of the bioactive ceramics (i.e., Si-OH groups and Ca2+ ions) are useful as novel bone substitutes, because of their bioactivity and mechanical properties analogous to those of natural bone. In the present study, organic-inorganic nanohybrids were synthesized from hydroxyethylmethacrylate (HEMA) and methacryloxypropyltrimethoxysilane (MPS), as well as various calcium salts. Bioactivity of the synthesized hybrids was assessed in vitro by examining their acceptance of apatite deposition in simulated body fluid (Kokubo solution). The prepared hybrids formed apatite in Kokubo solution when they were modified with calcium chloride (CaCl2) at 5 or 10 mol% of the total of MPS and HEMA. Deposition of a kind of calcium phosphate was observed for the hybrids modified with calcium acetate (Ca(CH3COO)2), although it could not be identified with apatite. The addition of glycerol up to 10 mol% of the total of MPS and HEMA or water up to 20 mol% as plasticizers did not appreciably decrease the acceptance of apatite formation of the hybrids. These findings allow wide selectivity in the design of bioactive nanohybrids developed by organic modification of the Si-OH group and calcium ion through sol-gel processing. Such nanohybrids have potential as novel bone substitutes with both high bioactivity and high flexibility.

Animals↗

Characterization of particle- and vapor-phase organic fraction emissions from a heavy-duty diesel engine equipped with a particle trap and regeneration controls.

The effects of a ceramic particle trap on the chemical and biological character of the exhaust from a heavy-duty diesel engine have been studied during steady-state operation and during periods of trap regeneration. Phase I of this project involved developing and refining the methods using a Caterpillar 3208 engine, and Phase II involved more detailed experiments with a Cummins LTA10-300 engine, which met Federal 1988 particulate matter standards, and a ceramic particle trap with built-in regeneration controls. During the Phase I experiments, samples wee collected at the Environmental Protection Agency (EPA)* steady-state mode 4 (50% load at intermediate speed). Varying the dilution ratio to obtain a constant filter-face temperature resulted in less variability in total particulate matter (TPM), particle-associated soluble organic fraction (SOF), solids (SOL), and polynuclear aromatic hydrocarbon (PAH) levels than sampling with a constant dilution ratio and allowing filter-face temperature to vary. A modified microsuspension Ames assay detected mutagenicity in the SOF samples, and in the semivolatile organic fraction extracted from XAD-2 resin (XAD-2 resin organic component, XOC) with at least 10 times less sample mass than the standard plate incorporation assay. Measurement techniques for PAH and nitro-PAH in the SOF and XOC also were developed during this portion of the project. For the Phase II work, two EPA steady-state rated speed modes were selected: mode 11 (25% load) and mode 9 (75% load). With or without the trap, filter-face temperatures were kept at 45 degrees +/- 2 degrees C, nitrogen dioxide (NO2) levels less than 5 parts per million (ppm), and sampling times less than 60 minutes. Particle sizes were determined using an electrical aerosol analyzer. Similar sampling methods were used when the trap was regenerated, except that a separate dilution tunnel and sampling system was designed and built to collect all of the regeneration emissions. The SOF and XOC were extracted from their collection media with dichloromethane. Levels of 12 PAH and nitro-PAH compounds with known biological effects were determined using high-pressure liquid chromatography with fluorescence detection. Two additional dinitro-PAHs were analyzed semi-quantitatively, and several representative SOF and XOC samples were screened for three additional PAH compounds. Mutagenicity was assessed using the modified Ames microsuspension assay.(ABSTRACT TRUNCATED AT 400 WORDS)

Air Pollutants↗

Partial biomimetic reconstitution of avian eggshell formation.

The avian eggshell is a biocomposite ceramic consisting of minute amounts of organic matrix and a crystalline calcium carbonate (calcite) filler. It is formed by a well regulated spatio-temporal assembling process, where extracellular matrix proteins, especially the sulfated glycosaminoglycan anionic sites of specific proteoglycans, have been involved in nucleation and growth of the inorganic crystalline phase. Together with such extracellular matrix molecules, the activity of carbonic anhydrase, is crucial for the normal eggshell formation. Here, we studied the effect of dermatan sulfate and carbonic anhydrase on the in vitro calcification of non-mineralized eggshell membrane-mammillae substrate at different pH and incubation times. Crystal morphology was analyzed by scanning electron microscopy. Crystal nucleation and growth was delayed at lower pH. Dermatan sulfate modified crystal morphology producing aggregates of large calcite crystals exhibiting a columnar morphology, contributing to the eggshell texture development. Carbonic anhydrase increased the velocity of crystal growth and eventually contributed to the fusion of the crystal aggregates to each other. Although, the effect of other macromolecules could not be ruled out, the combinatory effect of proteoglycans and carbonic anhydrase seems to be important for the control of eggshell formation.

Animals↗