Physical Conditions of Fast Glacier Flow: 3. Seasonally-Evolving Ice Deformation on Store Glacier, West Greenland.

Temporal variations in ice sheet flow directly impact the internal structure within ice sheets through englacial deformation. Large-scale changes in the vertical stratigraphy within ice sheets have been previously conducted on centennial to millennial timescales; however, intra-annual changes in the morphology of internal layers have yet to be explored. Over a period of 2 years, we use autonomous phase-sensitive radio-echo sounding to track the daily displacement of internal layers on Store Glacier, West Greenland, to millimeter accuracy. At a site located ∼30 km from the calving terminus, where the ice is ∼600 m thick and flows at ∼700 m/a, we measure distinct seasonal variations in vertical velocities and vertical strain rates over a 2-year period. Prior to the melt season (March-June), we observe increasingly nonlinear englacial deformation with negative vertical strain rates (i.e., strain thinning) in the upper half of the ice column of approximately -0.03 a-1, whereas the ice below thickens under vertical strain reaching up to +0.16 a-1. Early in the melt season (June-July), vertical thinning gradually ceases as the glacier increasingly thickens. During late summer to midwinter (August-February), vertical thickening occurs linearly throughout the entire ice column, with strain rates averaging 0.016 a-1. We show that these complex variations are unrelated to topographic setting and localized basal slip and hypothesize that this seasonality is driven by far-field perturbations in the glacier's force balance, in this case generated by variations in basal hydrology near the glacier's terminus and propagated tens of kilometers upstream through transient basal lubrication longitudinal coupling.

Synthesis and Assay of SIRT1-Activating Compounds.

The NAD(+)-dependent deacetylase SIRT1 plays key roles in numerous cellular processes including DNA repair, gene transcription, cell differentiation, and metabolism. Overexpression of SIRT1 protects against a number of age-related diseases including diabetes, cancer, and Alzheimer's disease. Moreover, overexpression of SIRT1 in the murine brain extends lifespan. A number of small-molecule sirtuin-activating compounds (STACs) that increase SIRT1 activity in vitro and in cells have been developed. While the mechanism for how these compounds act on SIRT1 was once controversial, it is becoming increasingly clear that they directly interact with SIRT1 and enhance its activity through an allosteric mechanism. Here, we present detailed chemical syntheses for four STACs, each from a distinct structural class. Also, we provide a general protocol for purifying active SIRT1 enzyme and outline two complementary enzymatic assays for characterizing the effects of STACs and similar compounds on SIRT1 activity.

Dynamics of impurity attraction and repulsion of an intrinsic localized mode in a driven 1-D cantilever array.

Both low frequency and high frequency impurity modes have been produced in a SiN micromechanical cantilever array by illumination with either an infrared or visible laser. When such laser-induced impurities are placed near a driven intrinsic localized mode (ILM), it is either repelled or attracted. By measuring the linear response spectrum for these two cases, it was found that vibrational hopping of the ILM takes place when the natural frequency of the ILM and an intrinsic even symmetry linear local mode are symmetrically located about the driven ILM frequency so that parametric excitation of these two linear modes is enhanced, amplifying the lateral motion of the ILM. Numerical simulations are consistent with these signature findings. It is also demonstrated that the correct sign of the observed interaction can be found with a harmonic lattice-impurity model but the magnitude of the effect is enhanced in a nonlinear lattice.

Switching dynamics and linear response spectra of a driven one-dimensional nonlinear lattice containing an intrinsic localized mode.

An intrinsic localized mode (ILM) represents a localized vibrational excitation in a nonlinear lattice. Such a mode will stay in resonance as the driver frequency is changed adiabatically until a bifurcation point is reached, at which point the ILM switches and disappears. The dynamics behind switching in such a many body system is examined here through experimental measurements and numerical simulations. Linear response spectra of a driven micromechanical array containing an ILM were measured in the frequency region between two fundamentally different kinds of bifurcation points that separate the large amplitude ILM state from the two low amplitude vibrational states. Just as a natural frequency can be associated with a driven harmonic oscillator, a similar natural frequency has been found for a driven ILM via the beat frequency between it and a weak, tunable probe. This finding has been confirmed using numerical simulations. The behavior of this nonlinear natural frequency plays important but different roles as the two bifurcation points are approached. At the upper transition its frequency coalesces with the driver and the resulting bifurcation is very similar to the saddle-node bifurcation of a single driven Duffing oscillator, which is treated in an Appendix. The lower transition occurs when the four-wave mixing partner of the natural frequency of the ILM intersects the topmost extended band mode of the same symmetry. The properties of linear local modes associated with the driven ILM are also identified experimentally for the first time and numerically but play no role in these transitions.

Experimental observation of the bifurcation dynamics of an intrinsic localized mode in a driven 1D nonlinear lattice.

Linear response spectra of a driven intrinsic localized mode in a micromechanical array are measured as it approaches two fundamentally different kinds of bifurcation points. A linear phase mode associated with this autoresonant state softens in frequency and its amplitude grows as the upper frequency bifurcation point is approached, similar to the soft-mode kinetic transition for a single driven Duffing resonator. A lower frequency bifurcation point occurs when the four-wave-mixing partner of this same phase mode intercepts the top of the extended wave branch, initiating a second kinetic transition process.

Glasslike two-level systems in minimally disordered mixed crystals.

THz spectroscopy is used to identify a broad distribution of two-level systems, characteristic of glasses, in the substitutional monatomic mixed crystal systems, Ba(1-x)Ca(x)F(2) and Pb(1-x)Ca(x)F(2). In these minimally disordered systems, two-level behavior, which was not previously known to occur, begins at a specific CaF(2) concentration. The concentration dependence, successfully modeled using the statistics of the impurity distribution in the lattice, points to a collective dopant tunneling mechanism.

Study of intrinsic localized vibrational modes in micromechanical oscillator arrays.

Intrinsic localized modes (ILMs) have been observed in micromechanical cantilever arrays, and their creation, locking, interaction, and relaxation dynamics in the presence of a driver have been studied. The micromechanical array is fabricated in a 300 nm thick silicon-nitride film on a silicon substrate, and consists of up to 248 cantilevers of two alternating lengths. To observe the ILMs in this experimental system a line-shaped laser beam is focused on the 1D cantilever array, and the reflected beam is captured with a fast charge coupled device camera. The array is driven near its highest frequency mode with a piezoelectric transducer. Numerical simulations of the nonlinear Klein-Gordon lattice have been carried out to assist with the detailed interpretation of the experimental results. These include pinning and locking of the ILMs when the driver is on, collisions between ILMs, low frequency excitation modes of the locked ILMs and their relaxation behavior after the driver is turned off.

Observation of locked intrinsic localized vibrational modes in a micromechanical oscillator array.

The nonlinear vibrational properties of a periodic micromechanical oscillator array have been measured. For sufficiently large amplitude of the driver, the optic mode of the di-element cantilever array becomes unstable and breaks up into excitations ranging over only a few cells. A driver-induced locking effect is observed to eternalize some of these intrinsic localized modes so that their amplitudes become fixed and the modes become spatially pinned.

Evolution of enzymatic activities in the enolase superfamily: functional assignment of unknown proteins in Bacillus subtilis and Escherichia coli as L-Ala-D/L-Glu epimerases.

The members of the mechanistically diverse enolase superfamily catalyze different overall reactions by using a common catalytic strategy and structural scaffold. In the muconate lactonizing enzyme (MLE) subgroup of the superfamily, abstraction of a proton adjacent to a carboxylate group initiates reactions, including cycloisomerization (MLE), dehydration [o-succinylbenzoate synthase (OSBS)], and 1,1-proton transfer (catalyzed by an OSBS that also catalyzes a promiscuous N-acylamino acid racemase reaction). The realization that a member of the MLE subgroup could catalyze a 1,1-proton transfer reaction, albeit poorly, led to a search for other enzymes which might catalyze a 1,1-proton transfer as their physiological reaction. YcjG from Escherichia coli and YkfB from Bacillus subtilis, proteins of previously unknown function, were discovered to be L-Ala-D/L-Glu epimerases, although they also catalyze the epimerization of other dipeptides. The values of k(cat)/K(M) for L-Ala-D/L-Glu for both proteins are approximately 10(4) M(-1) s(-1). The genomic context and the substrate specificity of both YcjG and YkfB suggest roles in the metabolism of the murein peptide, of which L-Ala-D-Glu is a component. Homologues possessing L-Ala-D/L-Glu epimerase activity have been identified in at least two other organisms.

Glycopeptide antibiotic biosynthesis: enzymatic assembly of the dedicated amino acid monomer (S)-3,5-dihydroxyphenylglycine.

Four proteins, DpgA-D, required for the biosynthesis by actinomycetes of the nonproteinogenic amino acid monomer (S)-3,5-dihydroxyphenylglycine (Dpg), that is a crosslinking site in the maturation of vancomycin and teicoplanin antibiotic scaffolds, were expressed in Escherichia coli, purified in soluble form, and assayed for enzymatic activity. DpgA is a type III polyketide synthase, converting four molecules of malonyl-CoA to 3,5-dihydroxyphenylacetyl-CoA (DPA-CoA) and three free coenzyme A (CoASH) products. Almost no turnover was observed for DpgA until DpgB was added, producing a net k(cat) of 1-2 min(-1) at a 3:1 ratio of DpgB:DpgA. Addition of DpgD gave a further 2-fold rate increase. DpgC had the unusual catalytic capacity to convert DPA-CoA to 3,5-dihydroxyphenylglyoxylate, which is a transamination away from Dpg. DpgC performed a net CH(2) to C=O four-electron oxidation on the Calpha of DPA-CoA and hydrolyzed the thioester linkage with a k(cat) of 10 min(-1). Phenylacetyl-CoA was also processed, to phenylglyoxylate, but with about 500-fold lower k(cat)/K(M). DpgC showed no activity in anaerobic incubations, suggesting an oxygenase function, but had no detectable bound organic cofactors or metals. A weak enoyl-CoA hydratase activity was detected for both DpgB and DpgD.

Tailoring enzymes that modify nonribosomal peptides during and after chain elongation on NRPS assembly lines.

Nonribosomal peptide synthetases are large enzyme complexes that synthesize a variety of peptide natural products through a thiotemplated mechanism. Assembly of the peptides proceeds through amino acid loading, amide-bond formation and chain translocation, and finally thioester lysis to release the product. The final products are often heavily modified, however, through methylation, epimerization, hydroxylation, heterocyclization, oxidative cross-linking and attachment of sugars. These activities are the province of specialized enzymes (either embedded in the multidomain nonribosomal peptide synthetase structure or standalone).

Aminoacyl-S-enzyme intermediates in beta-hydroxylations and alpha,beta-desaturations of amino acids in peptide antibiotics.

Many of the alpha-amino acids found in proteins are shunted into microbial secondary metabolism to form peptide antibiotics by specific oxidation, including hydroxylation, at the beta carbon. Examples for the enzymatic hydroxylation of tyrosine and histidine and for desaturation of proline during covalent attachment as aminoacyl-S-pantetheinyl enzyme intermediates suggest a general strategy in peptide antibiotic biosynthesis.

Community environmental health assessment in Peru's desert hills and rainforest.

Peru's expanding population and rapid urbanization--a result of migration to its largest cities--have stressed the country's public services infrastructure and the provision of public health and environmental health services. In response, the Ministry of Health established the General Directorate of Environmental Health (DIGESA), the branch charged with assuring adequate environmental health services to populations in rural and urban areas. The magnitude of the environmental health problems in peri-urban settlements, however, has exceeded the capacity of DIGESA to respond. The Urban Environmental Health Project is an effort to develop the ability of local communities to address these problems

Evolution of enzymatic activities in the enolase superfamily: identification of the general acid catalyst in the active site of D-glucarate dehydratase from Escherichia coli.

D-Glucarate dehydratase from Escherichia coli (GlucD), a member of the enolase superfamily, catalyzes the dehydration of both D-glucarate and L-idarate to form 5-keto-4-deoxy-D-glucarate (KDG). Previous mutagenesis and structural studies identified Lys 207 and the His 339-Asp 313 dyad as the general basic catalysts that abstract the C5 proton from L-idarate and D-glucarate, respectively, thereby initiating the reaction by formation of a stabilized enediolate anion intermediate [Gulick, A. M., Hubbard, B. K., Gerlt, J. A., and Rayment, I. (2000) Biochemistry 39, 4590-4602]. The vinylogous elimination of the 4-OH group from this intermediate presumably requires a general acid catalyst. The structure of GlucD with KDG and 4-deoxy-D-glucarate bound in the active site revealed that only His 339 and Asn 341 are proximal to the presumed position of the 4-OH leaving group. The N341D and N341L mutants of GlucD were constructed and subjected to both mechanistic and structural analyses. The N341L but not N341D mutant catalyzed the dehydrofluorination of 4-deoxy-4-fluoro-D-glucarate, demonstrating that in this mutant the initial proton abstraction from C5 can be decoupled from elimination of the leaving group from C4. The kinetic properties and structures of these mutants suggest that either Asn 341 participates in catalysis as the general acid that facilitates the departure of the 4-leaving group or is essential for proper positioning of His 339. In the latter scenario, His 339 would function not only as the general base that abstracts the C5 proton from D-glucarate but also as the general acid that catalyzes both the departure of the 4-OH group and the stereospecific incorporation of solvent hydrogen with retention of configuration to form the KDG product. The involvement of a single functional group in this reaction highlights the plasticity of the active site design in members of the enolase superfamily.

Molecular cloning and sequence analysis of the complestatin biosynthetic gene cluster.

Streptomyces lavendulae produces complestatin, a cyclic peptide natural product that antagonizes pharmacologically relevant protein-protein interactions including formation of the C4b,2b complex in the complement cascade and gp120-CD4 binding in the HIV life cycle. Complestatin, a member of the vancomycin group of natural products, consists of an alpha-ketoacyl hexapeptide backbone modified by oxidative phenolic couplings and halogenations. The entire complestatin biosynthetic and regulatory gene cluster spanning ca. 50 kb was cloned and sequenced. It consisted of 16 ORFs, encoding proteins homologous to nonribosomal peptide synthetases, cytochrome P450-related oxidases, ferredoxins, nonheme halogenases, four enzymes involved in 4-hydroxyphenylglycine (Hpg) biosynthesis, transcriptional regulators, and ABC transporters. The nonribosomal peptide synthetase consisted of a priming module, six extending modules, and a terminal thioesterase; their arrangement and domain content was entirely consistent with functions required for the biosynthesis of a heptapeptide or alpha-ketoacyl hexapeptide backbone. Two oxidase genes were proposed to be responsible for the construction of the unique aryl-ether-aryl-aryl linkage on the linear heptapeptide intermediate. Hpg, 3,5-dichloro-Hpg, and 3,5-dichloro-hydroxybenzoylformate are unusual building blocks that repesent five of the seven requisite monomers in the complestatin peptide. Heterologous expression and biochemical analysis of 4-hydroxyphenylglycine transaminon confirmed its role as an aminotransferase responsible for formation of all three precursors. The close similarity but functional divergence between complestatin and chloroeremomycin biosynthetic genes also presents a unique opportunity for the construction of hybrid vancomycin-type antibiotics.

Deoxysugars in glycopeptide antibiotics: enzymatic synthesis of TDP-L-epivancosamine in chloroeremomycin biosynthesis.

The 2,3,6-trideoxysugar l-epivancosamine is the terminal sugar added to the aglycone scaffold in chloroeremomycin, a member of the vancomycin family of glycopeptide antibiotics. Five proteins from the chloroeremomycin biosynthetic cluster, ORF14 and ORF23 to ORF26, have been expressed heterologously in Escherichia coli and purified to near homogeneity, and each has been characterized for an enzymatic activity. These five enzymes reconstitute the complete biosynthesis of TDP-l-epivancosamine from TDP-4-keto-6-deoxy-d-glucose. This process involves C-2 deoxygenation, C-3 amination and methylation, C-5 epimerization, and C-4 ketoreduction. Intermediates and the final product of this pathway have been identified by mass spectrometry and NMR. The pathway established here represents the complete in vitro reconstitution of an unusual sugar for an important class of antibiotics and sets the groundwork for future combinatorial biosynthesis for new bioactive compounds.

Pathologic findings in late endophthalmitis after glaucoma filtering surgery.

OBJECTIVE: To report the clinicopathologic features of four eyes enucleated for late-onset bleb-related endophthalmitis. STUDY DESIGN: Retrospective case series. MATERIALS: Four enucleated eyes. METHODS: The clinical and histopathologic features of four patients who underwent enucleation for late-onset endophthalmitis after glaucoma filtering surgery were reviewed. RESULTS: The eyes were enucleated for endophthalmitis one to five years after trabeculectomy. Two of the four eyes had trabeculectomy with adjunctive mitomycin-C. All four eyes had streptococci cultured from the aqueous and/or vitreous. Common pathologic features included inflammation involving the anterior segment, lens and choroid. One eye exhibited focal granulomatous uveitis. CONCLUSIONS: Late-onset endophthalmitis after glaucoma filtering surgery is often due to streptococcal species and rapidly progresses over a few days. Phacoanaphylaxis with associated granulomatous uveitis may contribute to the poor prognosis in this setting.

The effects of uncontrolled hyperglycemia on thrombosis and formation of neointima after coronary stent placement in a novel diabetic porcine model of restenosis.

BACKGROUND: Results of recent clinical studies suggest that patients with diabetes mellitus have a higher than normal rate of restenosis after percutaneous transluminal coronary angioplasty or coronary stenting. The mechanism for this exaggerated neointimal response is not known. OBJECTIVES: To determine the technical feasibility of a model of in-stent restenosis in swine with streptozotocin-induced hyperglycemia and to compare the late arterial responses to injury induced by placement of oversized coronary stents in diabetic and nondiabetic animals. METHODS: Eighteen 25-40 kg castrated male or intact female Yucatan miniature swine aged 6 months were obtained from a commercial supplier. Twelve of the miniature swine were randomly selected for intravenous treatment with 125 mg/kg streptozotocin to induce a hyperglycemic state. Twelve weeks after treatment, all animals underwent placement of oversized balloon-expandable stainless steel stents in the coronary arteries. After 28 days, histomorphometric analysis of the stented coronary arteries to determine the neointimal responses for the diabetic and nondiabetic animals was completed. RESULTS: Sudden death due to stent thrombosis occurred for five of 11 (45%) of the diabetic animals and none of the age-matched nondiabetic control animals (P=0.05). For histology after 28 days, the neointimal response was correlated to the extent of arterial injury for the diabetic (r=0.79, P < 0.0001) and nondiabetic (r=0.86, P < 0.0001) animals. The surviving diabetic animals had areas of neointimal (1.67 +/- 0.74 mm2) and percentages of in-stent stenosis (28 +/- 14) similar to those of the nondiabetic swine (1.36 +/- 0.40 mm2, P=0.26; 22 +/- 6, P=0.17). Multiple regression analysis also demonstrated that arterial injury (P < 0.0001) alone, not hyperglycemia (P=0.237), was independently correlated to formation of neointima. CONCLUSIONS: Uncontrolled hyperglycemia results in greater than normal thrombosis after coronary-stent placement in swine with streptozotocin-induced diabetes. These data suggest that greater than normal early formation of thrombus rather than proliferation of smooth muscle cells contributes to restenosis after coronary stenting in patients with diabetes mellitus.

General traits of personality and affectivity as predictors of satisfaction in intimate relationships: evidence from self- and partner-ratings.

Self- and partner-ratings on trait affect and the Big Five were obtained from 74 married and 136 dating couples. The relationship satisfaction of each person (the "target") was correlated with four sets of ratings: (a) target's self-rated personality, (b) target's partner-rated personality, (c) partner's self-rated personality, and (d) partner's target-rated personality. Self- and partner-ratings of the target's personality yielded very similar results. Negative and positive affectivity were consistent predictors of satisfaction in both samples. Conscientiousness and agreeableness were reliably related to satisfaction in the dating couples, whereas extraversion consistently correlated with satisfaction in the married couples. These traits jointly predicted as much as 34% (self-ratings) and 26% (partner-ratings) of the variance in satisfaction. In contrast, the partner's personality played a lesser role in satisfaction.