Repair or Observation of Resin Margin Defects: Clinical Trial After Five Years.

OBJECTIVE: To assess the effectiveness of repair/resealing of stained composite margins as an alternative to controlled observation without treatment in a randomized clinical trial after five years. METHODS AND MATERIALS: Each patient recruited had from one to three composite restorations with visible margin discoloration. Initially, the characteristics of each defect were recorded with direct vision through a surgical microscope at 20× magnification. Clinical evaluation was done by two independent examiners using modified USPHS criteria for color, margin discoloration, and margin adaptation. Each restoration was then randomly assigned to a control or treatment group. Control restorations were observed yearly for the presence of recurrent caries; treatment restorations were resealed by exposing the margin with a » round bur, removing all interfacial stain, acid etching, placing an adhesive bonding agent, and a flowable composite to restore margin integrity. There were 152 patients recruited, with 360 restorations (180 control and 180 treatment). RESULTS: At five years, 104 patients were recalled (68%) with 271 restorations (76%): 136 untreated control and 135 resealed restorations. At that time, 61 restorations had been lost or replaced for nonrelated reasons. Clinical evaluation of the remaining 210 restorations determined penetrating discoloration (control = 81%, resealed = 46%) and margin crevice formation (control = 21%, resealed = 11%). Recurrent caries was diagnosed cumulatively in only six control and five treatment restorations (<5%). Microscopically, 49 control restorations (49%) and 36 resealed restorations (33%) had crevice formation. Discoloration was distributed as follows: 9% vs 47% with no discoloration, 30% vs 33% in the composite, 49% vs 18% in the interface, and 12% vs 2% in tooth structure. CONCLUSIONS: Resealing of restorations with margin discoloration reduced the occurrence of penetrating stain from 81% in controls to 46% in resealed margins and crevicing from 21% to 11% after five years. Both controlled observation and resealing of margins resulted in a similar very low incidence (<6%) of recurrent caries.

31P NMR studies of Clostridium thermocellum. Mechanism of end product inhibition by ethanol.

31P NMR studies of intact cells and perchloric acid extracts are used to investigate the effect of ethanol on the bioenergetics and glycolysis of Clostridium thermocellum, an anaerobic bacterium potentially useful for the single step conversion of biomass to ethanol. Whole cells suspended in phosphate buffer and given a carbon source (cellobiose) at 60 degrees C rapidly establish a pH gradient across the membrane that can be monitored by the chemical shifts of inorganic phosphate in the exterior buffer and in the cytoplasm. Peak intensities can be related to phosphate active transport rates. Wild type bacteria and cells grown in inhibiting concentrations of ethanol establish similar pH gradients, but with slower kinetics and slower phosphate transport rates for the cells adapted to growth in ethanol. Direct addition of ethanol does not affect the rate of pH gradient formation or phosphate transport. Thus, while ethanol does not directly affect processes for energy conservation carried out by the membrane, adaptation to ethanol does alter membrane functions such as phosphate transport. 31P NMR spectra of perchloric acid extracts show that when wild type cells are adapted to grow in inhibiting concentrations of ethanol and then energized with cellobiose, sugar phosphate content is increased and the steady state distribution of glycolytic intermediates is altered. Nucleotide triphosphate/nucleotide diphosphate ratios are unaltered in these cells. These results strongly indicate that in C. thermocellum growth inhibition by ethanol is related to a blockage in glycolysis.

Ethanol-induced changes in the membrane lipid composition of Clostridium thermocellum.

When ethanol is added to the growth medium of Clostridium thermocellum ATCC 27405 and C9, a different membrane composition is observed after the period of growth arrest. Changes in fatty acid composition and some unsaturated, branched hydrocarbons have been monitored by GLC-MS. There is a marked increase in normal and anteiso-branched fatty acids at the expense of isobranched fatty acids and an increase in short and unsaturated fatty acids. Thus, an adaptive response to growth in the presence of ethanol induces a membrane containing fatty acids with lower melting points and produces a more 'fluid' membrane. The suggestion is made that these membrane changes may be maladaptive to the performance of C. thermocellum.

Development of ethanol tolerance in Clostridium thermocellum: effect of growth temperature.

The growth of Clostridium thermocellum ATCC 27405 and of C9, an ethanol-resistant mutant of this strain, at different ethanol concentrations and temperatures was characterized. After ethanol addition, cultures continued to grow for 1 to 2 h at rates similar to those observed before ethanol was added and then entered a period of growth arrest, the duration of which was a function of the age of inocula. After this period, cultures grew at an exponential rate that was a function of ethanol concentration. The wild-type strain showed a higher energy of activation for growth than the ethanol-tolerant derivative. The optimum growth temperature of the wild type decreased as the concentration of the ethanol challenge increased, whereas the optimum growth temperature for C9 remained constant. The results are discussed in terms of what is known about the effects of ethanol and temperature on membrane composition and fluidity.