Metric mechanics with nontrivial topology: Actuating irises, cylinders, and evertors.

Liquid crystal elastomers contract along their director on heating and recover on cooling, offering great potential as actuators and artificial muscles. If a flat sheet is programed with a spatially varying director pattern, then it will actuate into a curved surface, allowing the material to act as a strong machine such as a grabber or lifter. Here we study the actuation of programed annular sheets which, owing to their central hole, can sidestep constraints on area and orientation. We systematically catalog the set of developable surfaces encodable via axisymmetric director patterns and uncover several qualitatively new modes of actuation, including cylinders, irises, and everted surfaces in which the inner boundary becomes the outer boundary after actuation. We confirm our designs with a combination of experiments and numerics. Many of our actuators can reattain their initial inner or outer radius upon completing actuation, making them particularly promising, as they can avoid potentially problematic stresses in their activated state even when fixed onto a frame or pipe.

Shape-morphing living composites.

This work establishes a means to exploit genetic networks to create living synthetic composites that change shape in response to specific biochemical or physical stimuli. Baker's yeast embedded in a hydrogel forms a responsive material where cellular proliferation leads to a controllable increase in the composite volume of up to 400%. Genetic manipulation of the yeast enables composites where volume change on exposure to l-histidine is 14× higher than volume change when exposed to d-histidine or other amino acids. By encoding an optogenetic switch into the yeast, spatiotemporally controlled shape change is induced with pulses of dim blue light (2.7 mW/cm2). These living, shape-changing materials may enable sensors or medical devices that respond to highly specific cues found within a biological milieu.

Dual-responsive, shape-switching bilayers enabled by liquid crystal elastomers.

Materials that change shape are attractive candidates to replace traditional actuators for applications with power or size restrictions. In this work, we design a polymeric bilayer that changes shape in response to both heat and water by the incorporation of a water-responsive hydrophilic polymer with a heat-responsive liquid crystal elastomer. The distinct shape changes based on stimulus are controlled by the molecular order, and consequently the anisotropic modulus, of a liquid crystal elastomer. In response to water, the hydrophilic polymer layer expands, bending the bilayer along the path dictated by the anisotropic modulus of the liquid crystal elastomer layer, which is approximately 5 times higher along the molecular orientation than in perpendicular directions. We demonstrate that by varying the direction of this stiffer axis in LCE films, helical pitch of the swollen bilayer can be controlled from 0.1 to 20 mm. By spatially patterning the stiffer axis with a resolution of 900 µm2, we demonstrate bilayers that fold and bend based on the pattern within the LCE. In response to heat, the liquid crystal elastomer contracts along the direction of molecular order, and when this actuation is constrained by the hydrophilic polymer, this contraction results in a 3D shape that is distinct from the shape seen in water. Furthermore, by using the vitrification of the dry hydrophilic polymer this 3D shape can be retained in the bilayer after cooling. By utilizing sequential exposure to heat and water, we can drive the initially flat bilayer to reversibly shift between 3D shapes.

Post-Polymerization Crosslinked Polyurethane Shape-Memory Polymers.

Novel urethane shape-memory polymers (SMPs) of significant industrial relevance have been synthesized and characterized. Chemically crosslinked SMPs have traditionally been made in a one-step polymerization of monomers and crosslinking agents. However, these new post-polymerization crosslinked SMPs can be processed into complex shapes by thermoplastic manufacturing methods and later crosslinked by heat exposure or by electron beam irradiation. Several series of linear, olefinic urethane polymers were made from 2-butene-1,4-diol, other saturated diols, and various aliphatic diisocyanates. These thermoplastics were melt-processed into desired geometries and thermally crosslinked at 200°C or radiation crosslinked at 50 kGy. The SMPs were characterized by solvent swelling and extraction, differential scanning calorimetry (DSC), dynamic mechanical analysis (DMA), tensile testing, and qualitative shape-recovery analysis. Swelling and DMA results provided concrete evidence of chemical crosslinking, and further characterization revealed that the urethanes had outstanding mechanical properties. Key properties include tailorable transitions between 25 and 80°C, tailorable rubbery moduli between 0.2 and 4.2 MPa, recoverable strains approaching 100%, failure strains of over 500% at T(g), and qualitative shape-recovery times of less than 12 seconds at body temperature (37°C). Because of its outstanding thermo-mechanical properties, one polyurethane was selected for implementation in the design of a complex medical device. These post-polymerization crosslinked urethane SMPs are an industrially relevant class of highly processable shape-memory materials.

The wavelet-based cluster analysis for temporal gene expression data.

A variety of high-throughput methods have made it possible to generate detailed temporal expression data for a single gene or large numbers of genes. Common methods for analysis of these large data sets can be problematic. One challenge is the comparison of temporal expression data obtained from different growth conditions where the patterns of expression may be shifted in time. We propose the use of wavelet analysis to transform the data obtained under different growth conditions to permit comparison of expression patterns from experiments that have time shifts or delays. We demonstrate this approach using detailed temporal data for a single bacterial gene obtained under 72 different growth conditions. This general strategy can be applied in the analysis of data sets of thousands of genes under different conditions.

Three telomerases with completely non-telomeric template replacements are catalytically active.

Telomerase is a reverse transcriptase minimally composed of a reverse transcriptase protein subunit and an internal RNA component that contains the templating region. Point mutations of template RNA bases can cause loss of enzymatic activity, reduced processivity and misincorporation in vitro. Here we report the first complete replacement of the nine base TETRAHYMENA: thermophila telomerase templating region in vivo with non-telomeric sequences. Rather than ablating telomerase activity, three such replaced telomerases (U9, AUN and AU4) were effective in polymerization in vitro. In vivo, the AU4 and AUN genes caused telomere shortening. We demonstrated the fidelity of the AUN and U9 telomerases in vitro and utilized AUN telomerase to demonstrate that 5' end primer recognition by telomerase is independent of template base pairing. However, the mutant AUN template telomerase catalyzed an abnormal DNA cleavage reaction. For these U-only and AU- substituted templates, we conclude that base-specific interactions between the telomerase template and protein (or distant parts of the RNA) are not absolutely required for the minimal core telomerase functions of nucleotide addition and base discrimination.

Cross-tolerance between analgesia produced by xylazine and selective opioid receptor subtype treatments.

Opioid receptor agonists produce analgesia through multiple systems activated by stimulation of mu(1), mu(2), delta(1), delta(2) and kappa(1) opioid receptors. Morphine analgesia is modulated by stimulation of alpha(2) adrenoceptors. To understand how multiple opioid analgesic systems interact with alpha(2)-adrenoceptor systems, analgesic cross-tolerance between the alpha(2) adrenoceptor agonist xylazine and opioid receptor agonists was studied using the mouse tail-flick assay. Mice received either xylazine (20 mg/kg, s.c.) or saline (1 ml/kg) for five days. On day six, mice received a dose of s.c. xylazine, i.c.v. [D-Ala(2),MePhe(4),Gly(ol)(5)]enkephalin (DAMGO), i.t. Tyr-Pro-Trp-Gly-NH(2) (Tyr-W-MIF-1), i.c.v. or i.t. [D-Pen(2),D-Pen(5)]enkephalin (DPDPE), i.t. [D-Ala(2)]deltorphin II (deltorphin II), or s.c. trans-(+/-)-3, 4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl-cyclohexyl] benzeneacetamide (U50,488). Xylazine tolerant mice required 4. 57-fold more xylazine to elicit the same response as saline treated animals and showed a 2.55-fold shift in i.c.v. DAMGO and a 3.37-fold shift in i.c.v. DPDPE antinociception. No cross-tolerance was seen with i.c.v. deltorphin II, i.t.Tyr-W-MIF-1, i.t. DPDPE, i.t. Tyr-W-MIF-1 or s.c. U50,488. These results implicate alpha(2) adrenoceptor systems in the modulation of supraspinal mu(1), and delta(1) opioid analgesic circuitry and raise the possibility that mu(2), delta(2) or kappa(1) opioid receptor agonists may be alternated with alpha(2) adrenoceptor agonists to minimize tolerance or treat opioid-tolerant patients.

Influence of malic acid supplementation on ruminal pH, lactic acid utilization, and digestive function in steers fed high-concentrate finishing diets.

Two trials were conducted to evaluate the influence of malic acid supplementation on ruminal fermentation. In Trial 1, six Holstein steers (300 kg) with ruminal cannulas were used in a crossover design experiment to study the influence of malic acid (MA) on ruminal metabolism during glucose-induced lactic acidosis. Treatments consisted of a 77% steam-flaked barley-based finishing diet supplemented to provide 0 or 80 g/d of MA. After a 13-d dietary adjustment period, 1 kg of glucose was infused into the rumen 1 h after the morning feeding. Ruminal pH was closely associated (R2 = .70) with ruminal DL-lactate concentration. Malic acid supplementation increased (P < .01) ruminal pH 3 h after the glucose infusion. However, there were no treatment effects (P > .10) on ruminal VFA molar proportions or ruminal and plasma DL-lactate concentrations. In Trial 2, four Holstein steers (150 kg) with cannulas in the rumen and proximal duodenum were used in a crossover design experiment to evaluate the influence of MA supplementation on characteristics of digestion. Treatments consisted of an 81% steam-flaked barley-based finishing diet supplemented to provide 0 or 80 g/d of MA. There were no treatment effects (P > .10) on ruminal and total tract digestion of OM, ADF, starch, and feed N or on ruminal microbial efficiency. Malic acid supplementation increased (P < .05) ruminal pH 2 h after feeding. As with Trial 1, there were no treatment effects (P > .10) on ruminal VFA and DL-lactate concentrations. We conclude that supplementation of high-grain finishing diets with MA may be beneficial in promoting a higher ruminal pH during periods of peak acid production without detrimental effects on ruminal microbial efficiency or starch, fiber, and protein digestion. There were no detectable beneficial effects of MA supplementation on ruminal and plasma lactic acid concentrations in cattle fed high-grain diets.

The telomere and telomerase: how do they interact?

The tandemly repeated DNA sequence of telomeres is typically specified by the ribonucleoprotein enzyme telomerase. Telomerase copies part of its intrinsic RNA moiety to make one strand of the telomeric repeat DNA. Recent work has led to the concept of a telomere homeostasis system. We have been studying two key physical components of this system: the telomere itself and telomerase. Mutating the template sequence of telomerase RNA caused various phenotypes: (1) mutating specific residues in the ciliate Tetrahymena and two yeasts showed that they are required for critical aspects of telomerase action; (2) certain mutated telomeric sequences caused a previously unreported phenotype, i.e. a strong anaphase block in Tetrahymena micronuclei; and (3) certain template mutations in the telomerase RNA gene of the yeast Kluyveromyces lactis led to unregulated telomere elongation, which in some cases was directly related to loss of binding to K. lactis Rap1p. Using K. lactis carrying alterations in the genes for Rap1p and other silencing components, we proposed a general model for telomere length homeostasis: namely, that the structure and DNA length of the DNA-protein complex that comprises the telomere are key determinants of telomerase access, and hence the frequency of action of telomerase, at the telomere.

The air-liquid interface and the pH and PCO2 of alkalinized local anaesthetic solutions.

The alkalinization of certain local anaesthetics with sodium bicarbonate hastens the onset of epidural analgesia. Increases in both the pH and PCO2 of the local anaesthetic are necessary to hasten onset. However, carbon dioxide can diffuse from local anaesthetic solutions following alkalinization with sodium bicarbonate and change both the pH and PCO2 of the mixture. This study examined changes in pH and PCO2 of three local anaesthetics reported to have a faster onset of analgesia following mixture with sodium bicarbonate and examined the effects of time and the local anaesthetic container air/liquid interface on the pH and PCO2 of the buffered local anaesthetic solutions. Bupivacaine 0.5%, lidocaine 2%, and chloroprocaine 2% were each buffered with sodium bicarbonate. The pH and PCO2 of each solution were measured at time 0 and at 5, 10, 15, 20, 30, 40, 50 and 60 min intervals. The solutions were placed in containers as follows: 30 ml in 40 ml containers, 10 ml in 40 ml containers, 10 ml in 13 ml containers, and 10 ml in polypropylene syringes. The pH and PCO2 increased following alkalinization but gradually decreased in all containers except in polypropylene syringes.

The air-liquid interface and the pH and PCO2 of alkalinized local anaesthetic solutions.

The alkalinization of certain local anaesthetics with sodium bicarbonate hastens the onset of epidural analgesia. Increases in both the pH and PCO2 of the local anaesthetic are necessary to hasten onset. However, carbon dioxide can diffuse from local anaesthetic solutions following alkalinization with sodium bicarbonate and change both the pH and PCO2 of the mixture. This study examined changes in pH and PCO2 of three local anaesthetics reported to have a faster onset of analgesia following mixture with sodium bicarbonate and examined the effects of time and the local anaesthetic container air/liquid interface on the pH and PCO2 of the buffered local anaesthetic solutions. Bupivacaine 0.5%, lidocaine 2%, and chloroprocaine 2% were each buffered with sodium bicarbonate. The pH and PCO2 of each solution were measured at time 0 and at 5, 10, 15, 20, 30, 40, 50 and 60 min intervals. The solutions were placed in containers as follows: 30 ml in 40 ml containers, 10 ml in 40 ml containers, 10 ml in 13 ml containers, and 10 ml in polypropylene syringes. The pH and PCO2 increased following alkalinization but gradually decreased in all containers except in polypropylene syringes.