Persistent thrombin generation in humans during specific thrombin inhibition with hirudin

BACKGROUND: The degree to which antithrombotic drugs suppress thrombin generation is unknown. Because hirudin, unlike antithrombin III, binds intravascular thrombin rapidly and selectively to yield a circulating inactive complex of 3- to 4-hour half-life, we used intravenous hirudin in humans to investigate the course of thrombin generation during and early after anticoagulation with this potent, direct antithrombin. METHODS AND RESULTS: Intravascular thrombin was measured with an ELISA for the thrombin-hirudin complex formed during and for 18 hours after stopping a 6-hour infusion of hirudin at 0.1, 0.2, and 0.3 mg.kg-1.h-1 in three groups of six patients each. With free hirudin in 20- to 10,000-fold molar excess of thrombin and peak activated partial thromboplastin times of 2.3 to 3.0 times baseline, mean plasma thrombin-hirudin complex increased from 794 +/- 85 pg/mL (mean +/- SEM) 15 minutes after the start of the infusion to 1617 +/- 151 pg/mL at 6 hours of infusion to 2667 +/- 654 pg/mL at 24 hours. During the 24-hour observation period, plasma concentration of fragment 1.2 (the peptide released during conversion of prothrombin to thrombin) never fell below baseline but rather increased transiently during the hirudin infusion. Plasma concentrations of thrombin-antithrombin III complex (in ng/mL) decreased from 4.34 +/- 0.40 at baseline to 1.64 +/- 0.13 at 6 hours (P < .001) and gradually increased after stopping the infusion to 5.7 +/- 0.87 at 24 hours (nonsignificant compared with baseline). CONCLUSIONS: Measurement of thrombin-hirudin complex may be used as a marker of thrombin generation in humans. Persistent accumulation of thrombin-hirudin complex and generation of fragment 1.2 during and after completion of potent anticoagulation with hirudin suggest thrombin generation is not blocked by high-affinity thrombin inhibition. The persistent formation of thrombin during declining plasma levels of hirudin may contribute to the pathogenesis of rethrombosis early after antithrombin therapy or during inadequate anticoagulation.     

Role of diameter differences among follicles in selection of a future dominant follicle in mares

Follicles > or = 5 mm were ablated in pony mares by a transvaginal ultrasound-guided technique on Day 10 (ovulation = Day 0). Follicle emergence (at 15 mm, experiment 1; at 6 mm, experiment 2) and development of the new wave was monitored by transrectal ultrasound. Deviation was defined as the beginning of a marked difference in growth rates between the two largest follicles. In experiment 1, mares were grouped (n = 4 per group) into controls, ablation-controls (ablations at Day 10 only), and a two-follicle model (periodic ablation sessions so that only the two largest follicles developed). There were no significant indications that the two-follicle model altered follicle diameters, growth rates, or time intervals of the two retained follicles at or between events (follicle emergence, deviation, and ovulation). In experiment 2, the two-follicle model (n = 14) was used for follicle and hormonal characterization and hypothesis testing, without the tedious and error-prone necessity for tracking many (e.g., 20) individual follicles. The future dominant follicle emerged a mean of 1 day earlier (p < 0.008) than the future subordinate follicle, the growth rates for the two follicles between emergence and deviation (6 days later) did not differ, and the dominant follicle was larger at the beginning of deviation (23.1 +/- 0.8 mm versus 19.6 +/- 0.9 mm; p < 0.0001). Mean FSH and LH concentrations increased (p < 0.05) concomitantly from emergence of the future dominant follicle and peaked 3 days later when the follicle was a mean of 13 mm. Thereafter, the two hormones disassociated until ovulation: FSH decreased and LH increased. Results supported the hypothesis that the future dominant follicle has an early size advantage over future subordinate follicles and indicated that the advantage was present as early as 6 days before deviation.     

Acetophenone as an Anti-attractant for the Western Pine Beetle, <Emphasis Type="Italic">Dendroctonus Brevicomis</Emphasis> LeConte (Coleoptera: Scolytidae)

Host location and colonization by bark beetles is dependent upon the relative and absolute amounts of attractant and antiattractant compounds available. Many investigations have lead to use of antiattractants for the management of these pests and have been especially focused on verbenone. However, recent studies have identified new antiattractants for several species of bark beetles. We report results of recent investigations of the response of western pine beetle, Dendroctonus brevicomis LeConte, to two recently identified antiattractants, acetophenone, and fenchyl alcohol, with verbenone as a standard of comparison, in northern California. Release of both acetophenone and verbenone resulted in significantly lower trap catches of D. brevicomis in aggregation pheromone-baited traps, while fenchyl alcohol was inactive. Acetophenone was the only antiattractant that did not reduce numbers of the most abundant predator of D. brevicomis, Temnochila chlorodia (Mannerheim), responding to the attractant pheromone of its prey. Aggregation pheromone-baited traps with acetophenone also had the highest predator/prey ratio. Our results suggest that acetophenone may be part of the intra- and interspecific interactions among sympatric species of bark beetles and may have application in their control.     

Nucleation and Growth Characteristics of Cavities during the Early Stages of Tensile Creep Deformation in a Superplastic Zirconia–20 wt% Alumina Composite

Constant-stress tensile creep experiments on a superplastic 3-mol%-yttria-stabilized tetragonal zirconia composite with 20 wt% alumina revealed that cavities nucleate relatively early during tensile deformation. The number of cavities nucleated increases with increasing imposed stress. The cavities nucleate at triple points associated largely with an alumina grain, and then grow rapidly in a cracklike manner to attain dimensions on the order of the grain facet size. It is suggested that coarser-grained superplastic ceramics exhibit lower ductility due to the ease in formation of such grain boundary facet-cracks and their interlinkage to form a macroscopic crack of critical dimensions.     

Additive-free hydrogelation of graphene oxide by ultrasonication

Graphene oxide hydrogels have been prepared by ultrasonication of precursor aqueous dispersions. The ultrasonication fractures the nanosheets, reducing their dimensions and exposing new sheet edges that do not possess the stabilizing carboxyl functional groups found along the edge of the as-prepared material. Ultrasonication does not affect the overall chemical functionality of the graphene oxide nanosheets, as spectra (carbon-13 nuclear magnetic resonance spectroscopy, infrared spectroscopy, and X-ray photoelectron spectroscopy) of samples before and after ultrasonication are nearly identical. Gelation is induced after only 30 min of ultrasonication to achieve a relatively weak gel with a shear modulus of 0.3 kPa; however, extension of ultrasonic treatment to 120 min yields a more robust hydrogel with a shear modulus of 1.6 kPa. Such enhancement in the gel’s physical properties can be attributed to the lack of stabilizing carboxyl groups on newly generated nanosheet fragments from the interior regions of the original nanosheets. As prepared, these hydrogels exhibit exceptionally low critical gelation concentrations ranging from ～0.050 to ～0.125 mg mL^?1 that can be tuned according to the extent of ultrasonic treatment.     

Polymers used to influence cell fate in 3D geometry: New trends

The extracellular matrix (ECM) is a hydrogel-like structure comprised of several different biopolymers, encompassing a wide range of biological, chemical, and mechanical properties. The composition, organization, and assembly of the ECM play a critical role in cell function. Cellular behavior is guided by interactions that occur between cells and their local microenvironment, and this interrelationship plays a significant role in determining physiological functions. Bioengineering approaches have been developed to mimic native tissue microenvironments by fabricating novel bioactive hydrogel scaffolds. This review explores material designs and fabrication approaches that are guiding the design of hydrogels as tissue engineered scaffolds. As the fundamental biology of the cellular microenvironment is often the inspiration for material design, the review focuses on modifications to control bioactive cues such as adhesion molecules and growth factors, and summarizes the current applications of biomimetic scaffolds that have been used in vitro as well as in vivo.     

Isolation of Non-methylene Interrupted or Acetylenic Fatty Acids from Seed Oils Using Semi-preparative Supercritical Chromatography

Preparative scale supercritical fluid chromatography was used for isolating and purifying uncommon non-methylene interrupted or acetylenic polyunsaturated fatty acids ethyl esters from seed oils. Fractionation of Biota orientalis seed oil ethyl esters was performed by supercritical fluid chromatography to obtain juniperonic acid [(5Z,11Z,14Z,17Z)-eicosa-5,11,14,17-tetraenoic acid], a non-methylene interrupted polyunsaturated fatty acid. Fractionation of sandalwood seed oil ethyl esters yielded ximenynic acid [(E)-octadec-11-en-9-ynoic acid], an acetylenic polyunsaturated fatty acid. The effects of CO₂ flow rate, column stationary phase and particle size were explored to optimize ximenynic and juniperonic ethyl ester recovery and purity from ethyl ester mixtures using online UV/Vis detection. Particle size, followed by the stationary phase, were found to be the most important parameters to achieving good separation. Under optimized conditions, ximenynic and juniperonic ethyl ester purities greater than 99 and 95%, respectively, were achieved in a one step process without co-solvent. The isolation and recovery of juniperonic acid from biota seed oil free fatty acids was also attempted. Using free fatty acids as the feed material, the non-methylene interrupted polyunsaturated sciadonic acid was also able to be separated from other compounds including juniperonic acid under some conditions, and gave an increase in concentration of more than 17 times.