Indirect selective laser sintering of apatite-wollostonite glass-ceramic.

This paper develops an indirect selective laser sintering (SLS) processing route for apatite-wollastonite (A-W) glass-ceramic, and shows that the processing route, which can create porous three-dimensional products suitable for bone implants or scaffolds, does not affect the excellent mechanical and biological properties of the glass-ceramic. 'Green parts' with fine integrity and well-defined shape have been produced from glass particles of single-size range or mixed-size ranges with acrylic binder in various ratios by weight. A subsequent heat treatment process has been developed to optimize the crystallization process, and an infiltration process has been explored to enhance mechanical strength. Three-point bending test results show flexural strengths of up to 102 MPa, dependent on porosity, and simulated body fluid (SBF) tests show that the laser sintered porous A-W has comparable biological properties to that of conventionally produced A-W.

Indirect selective laser sintering of an apatite-mullite glass-ceramic for potential use in bone replacement applications.

The feasibility of using indirect selective laser sintering (SLS) to produce parts from glass-ceramic materials for bone replacement applications has been investigated. A castable glass based on the system SiO2 x Al2O3 x P2O5 x CaO x CaF2 that crystallizes to a glass-ceramic with apatite and mullite phases was produced, blended with an acrylic binder, and processed by SLS. Green parts with good structural integrity were produced using a wide range of processing conditions, allowing both monolayer and multilayer components to be constructed. Following SLS the parts were post-processed to remove the binder and to crystallize fully the material, evolving the apatite and mullite phases. The parts were heated to 1200 degrees C using a number of different time-temperature profiles, following which the processed material was analysed by differential thermal analysis, X-ray diffraction, and scanning electron microscopy, and tested for flexural strength. An increase in strength was achieved by infiltrating the brown parts with a resorbable phosphate glass, although this altered the crystal phases present in the material.

Effects of marine reserves on adjacent fisheries.

Marine reserves have been widely promoted as conservation and fishery management tools. There are robust demonstrations of conservation benefits, but fishery benefits remain controversial. We show that marine reserves in Florida (United States) and St. Lucia have enhanced adjacent fisheries. Within 5 years of creation, a network of five small reserves in St. Lucia increased adjacent catches of artisanal fishers by between 46 and 90%, depending on the type of gear the fishers used. In Florida, reserve zones in the Merritt Island National Wildlife Refuge have supplied increasing numbers of world record-sized fish to adjacent recreational fisheries since the 1970s. Our study confirms theoretical predictions that marine reserves can play a key role in supporting fisheries.

Delayed effects of embryonic exposure of zebrafish (Danio rerio) to methylmercury (MeHg).

Since previous short-term bioassays of methylmercury (MeHg) indicated no morphological effects in zebrafish (Danio rerio) after embryonic exposures below 20 microg/l MeHg, studies were done to determine whether embryonic exposure to MeHg at lower concentrations would induce behavioral effects. Newly fertilized embryos were exposed to 0, 5, 10 or 15 microg MeHg/l for selected exposure durations: single day, multiple day or continuous exposure from fertilization through hatching. Larvae were maintained in an essential salt solution after hatching. Spontaneous swimming performance and prey capture experiments were conducted. Continuous embryonic exposure to 15 microg/l caused delayed mortality syndrome (DMS). These larvae hatched normally and appeared normal, but beginning at Day 3 post-hatch (ph), general activity was severely reduced and by Day 5 ph, larvae were completely moribund; many had faint heartbeats, severely enlarged body cavities and upward flexures of the spinal cord. Most of these larvae were dead by Day 6 ph. Multi- and single-day embryonic exposures to 15 microg/l caused reduced swimming activity and prey capture ability, and by Day 4 ph, these larvae also began to show signs of DMS. Continuous embryonic exposure to 10 microg/l significantly reduced spontaneous swimming activity, which did not improve after 5 days in clean water. Similar results were seen in larvae exposed during the last 24 h of embryonic development. Prey capture ability was also impaired in larvae exposed continuously to 10 microg/l, even after 4 days in clean water. Single-day exposures to 10 microg/l did not affect prey capture ability. Larvae from the 5-microg/l exposures were not significantly different from controls for either parameter. This study reinforces the idea that functional impairment is a more subtle response to developmental toxicants than mortality or the production of morphological defects.

Variations in the compressive strength of dental cements stored in ionic or acidic solutions.

The compressive strengths of various dental cements (a zinc polycarboxylate, a zinc phosphate, a glass-ionomer and two resin-modified glass ionomers, RMGICs) have been determined following storage in pure water, 0.9% sodium chloride solution or 20 mmol dm(-3) lactic acid solution for periods of time ranging from 24 h to 3 months. The glass-ionomer cement showed no differences between different storage solutions or at different storage times, whereas the zinc polycarboxylate, zinc phosphate and the resin-modified glass ionomer cements showed significant differences following storage in the solutions for 24 h compared with pure water. The zinc polycarboxylate cement was significantly weaker at 24 h in 0.9% NaCl and lactic acid than in pure water, whereas most of the other cements were significantly stronger in both 0.9% NaCl and lactic acid. One of the RMGICs (Vitremer luting, ex. 3M), however, was significantly stronger only in the NaCl solution, not in the lactic acid. In general, by 1 week, the strengths all reverted to being essentially the same as for specimens stored in pure water for most subsequent storage times, and did not change significantly on storage for up to 3 months. This effect of storage medium on the early strength has not been reported previously and since the media were chosen to model certain characteristics of natural saliva, the changes observed seem likely to occur in vivo. It is concluded that pure water is not the best medium for storing these cements if they are to behave as they do under clinical conditions.

Cytotoxic effects of dihydroorotase inhibitors upon human CCRF-CEM leukemia.

6-L-Thiodihydroorotate (TDHO) and 2-oxo-1,2,3,6-tetrahydropyrimidine-4,6-dicarboxylate (HDDP) are potent inhibitors of mammalian dihydroorotase in vitro (R. I. Christopherson, K. J. Schmalzl, E. Szabados, R. J. Goodridge, M. C. Harsanyi, M. E. Sant, E. M. Algar, J. E. Anderson, A. Armstrong, S. C. Sharma, W. A. Bubb, and S. D. Lyons, Biochemistry, 28: 463-470, 1989). Using human CCRF-CEM leukemia cells growing in culture, TDHO and HDDP as the free acids have 50% inhibitory concentration (IC50) values of 32 microM and greater than 1000 microM, respectively, whereas for TDHO methyl ester, the IC50 value is 25 microM, and for HDDP dimethyl ester, the IC50 value is 21 microM. These IC50 values were not affected by addition of dihydroorotate, uridine, or deoxycytidine to the culture medium. TDHO methyl ester (25 microM) had only slight inhibitory effects upon the dihydroorotase reaction of de novo pyrimidine biosynthesis in growing leukemia cells, cells arrested in G2 + M phases of the cell cycle. At 250 microM TDHO methyl ester, analysis of cell extracts by high-performance liquid chromatography showed that after 4 h carbamyl aspartate had accumulated from undetectable levels to 760 microM, whereas UTP decreased from 580 to 110 microM and CTP from 350 to 86 microM, indicating inhibition of dihydroorotase in growing leukemia cells. IMP accumulated from 63 to 350 microM, total guanylates increased while adenylates decreased, and the adenylate energy charge decreased from 0.91 to 0.69 after 4 h. The cellular concentration of 5-phosphoribosyl 1-pyrophosphate increased from 180 to 290 microM due to sparing from pyrimidine nucleotide biosynthesis resulting in complementary stimulation of the de novo purine pathway. HDDP dimethyl ester at concentrations of up to 250 microM had no discernable effect upon pyrimidine or purine nucleotide biosynthesis. At 25 microM HDDP-dimethyl ester, cells arrested in G2 + M phases initially, with accumulation of cells in G1/G0 at later times. These data suggest that the primary mechanisms of growth inhibition for TDHO and HDDP involve inhibition of cell cycle progression from late G2 or M phase to G1 phase and that blockade of the pyrimidine pathway by TDHO is a secondary effect found at higher concentrations.

Mercaptan and dicarboxylate inhibitors of hamster dihydroorotase.

In mammals, dihydroorotase is part of a trifunctional protein, dihydroorotate synthetase, which catalyzes the first three reactions of de novo pyrimidine biosynthesis. Dihydroorotase catalyzes the formation of a peptide-like bond between the terminal ureido nitrogen and the beta-carboxyl group of N-carbamyl-L-aspartate to yield heterocyclic L-dihydroorotate. A variety of evidence suggests that dihydroorotase may have a catalytic mechanism similar to that of a zinc protease [Christopherson, R. I., & Jones, M. E. (1980) J. Biol. Chem. 255, 3358-3370]. Tight-binding inhibitors of the zinc proteases, carboxypeptidase A, thermolysin, and angiotensin-converting enzyme have been synthesized that combine structural features of the substrates with a thiol or carboxyl group in an appropriate position to coordinate a zinc atom bound at the catalytic site. We have synthesized (4R)-2-oxo-6-thioxohexahydropyrimidine-4-carboxylate (L-6-thiodihydroorotate) and have found that this analogue is a potent competitive inhibitor of dihydroorotase with a dissociation constant (Ki) in the presence of excess Zn2+ ion of 0.17 +/- 0.02 microM at pH 7.4. The potency of inhibition by L-6-thiodihydroorotate in the presence of divalent metal ions decreases in the order Zn2+ greater than Ca2+ greater than Co2+ greater than Mn2+ greater than Ni2+; L-6-thiodihydroorotate alone is less inhibitory and has a Ki of 0.85 +/- 0.14 microM. 6-Thioorotate has a Ki of 82 +/- 8 microM which decreases to 3.8 +/- 1.4 microM in the presence of Zn2+. Zn2+ alone is a moderate inhibitor of dihydroorotase and does not enhance the potency of other inhibitors.(ABSTRACT TRUNCATED AT 250 WORDS)