Structural Health Monitoring of Fatigue Cracks for Steel Bridges with Wireless Large-Area Strain Sensors.

This paper presents a field implementation of the structural health monitoring (SHM) of fatigue cracks for steel bridge structures. Steel bridges experience fatigue cracks under repetitive traffic loading, which pose great threats to their structural integrity and can lead to catastrophic failures. Currently, accurate and reliable fatigue crack monitoring for the safety assessment of bridges is still a difficult task. On the other hand, wireless smart sensors have achieved great success in global SHM by enabling long-term modal identifications of civil structures. However, long-term field monitoring of localized damage such as fatigue cracks has been limited due to the lack of effective sensors and the associated algorithms specifically designed for fatigue crack monitoring. To fill this gap, this paper proposes a wireless large-area strain sensor (WLASS) to measure large-area strain fatigue cracks and develops an effective algorithm to process the measured large-area strain data into actionable information. The proposed WLASS consists of a soft elastomeric capacitor (SEC) used to measure large-area structural surface strain, a capacitive sensor board to convert the signal from SEC to a measurable change in voltage, and a commercial wireless smart sensor platform for triggered-based wireless data acquisition, remote data retrieval, and cloud storage. Meanwhile, the developed algorithm for fatigue crack monitoring processes the data obtained from the WLASS under traffic loading through three automated steps, including (1) traffic event detection, (2) time-frequency analysis using a generalized Morse wavelet (GM-CWT) and peak identification, and (3) a modified crack growth index (CGI) that tracks potential fatigue crack growth. The developed WLASS and the algorithm present a complete system for long-term fatigue crack monitoring in the field. The effectiveness of the proposed time-frequency analysis algorithm based on GM-CWT to reliably extract the impulsive traffic events is validated using a numerical investigation. Subsequently, the developed WLASS and algorithm are validated through a field deployment on a steel highway bridge in Kansas City, KS, USA.

Use of flexible sensor to characterize biomechanics of canine skin.

BACKGROUND: Suture materials and techniques are frequently evaluated in ex vivo studies by comparing tensile strengths. However, the direct measurement techniques to obtain the tensile forces in canine skin are not available, and, therefore, the conditions suture lines undergo is unknown. A soft elastomeric capacitor is used to monitor deformation in the skin over time by sensing strain. This sensor was applied to a sample of canine skin to evaluate its capacity to sense strain in the sample while loaded in a dynamic material testing machine. The measured strain of the sensor was compared with the strain measured by the dynamic testing machine. The sample of skin was evaluated with and without the sensor adhered. RESULTS: In this study, the soft elastomeric capacitor was able to measure strain and a correlation was made to stress using a modified Kelvin-Voigt model for the canine skin sample. The sensor significantly increases the stiffness of canine skin when applied which required the derivation of mechanical models for interpretation of the results. CONCLUSIONS: Flexible sensors can be applied to canine skin to investigate the inherent biomechanical properties. These sensors need to be lightweight and highly elastic to avoid interference with the stress across a suture line. The sensor studied here serves as a prototype for future sensor development and has demonstrated that a lightweight highly elastic sensor is needed to decrease the effect on the sensor/skin construct. Further studies are required for biomechanical characterization of canine skin.

UAV rapidly-deployable stage sensor with electro-permanent magnet docking mechanism for flood monitoring in undersampled watersheds.

The availability of historical flood data is vital in recognizing weather-related trends and outlining necessary precautions for at-risk communities. Flood frequency, magnitude, endurance, and volume are traditionally recorded using established streamgages; however, the material and installation costs allow only a few streamgages in a region, which yield a narrow data selection. In particular, stage, the vertical water height in a water body, is an important parameter in determining flood trends. This work investigates a low-cost, compact, rapidly-deployable alternative to traditional stage sensors that will allow for denser sampling within a watershed and a more detailed record of flood events. The package uses a HC-SR04 ultrasonic sensor to measure stage, onboard memory for recording flood events, and an electropermanet magnet (EPM) to enable Unmanned Aerial Vehicle (UAV) deployments. Optional modules for solar panels and wireless communication can also be added to extend package longevity or allow wireless control of the EPM. The stage sensor package was found to have a range of 0.02 to 4 m with a 6.9 mm accuracy and capable of a 6.4 day long deployment. With the total cost of production at 271.37 USD, it is a cheaper and more flexible alternative to traditional stage sensors that will enable dense sensor networks and rapid response to flooding events.

latheo encodes a subunit of the origin recognition complex and disrupts neuronal proliferation and adult olfactory memory when mutant.

The Drosophila latheo (lat) gene was identified in a behavioral screen for olfactory memory mutants. The original hypomorphic latP1 mutant (Boynton and Tully, 1992) shows a structural defect in adult brain. Homozygous lethal lat mutants lack imaginal discs, show little cell proliferation in the CNS of third instar larvae, and die as early pupae. latP1 was cloned, and all of the above mentioned defects of hypomorphic or homozygous lethal lat mutants were rescued with a lat+ transgene. lat encodes a novel protein with homology to a subunit of the origin recognition complex (ORC). Human and Drosophila LAT both associate with ORC2 and are related to yeast ORC3, suggesting that LAT functions in DNA replication during cell proliferation.

A domain of the even-skipped protein represses transcription by preventing TFIID binding to a promoter: repression by cooperative blocking.

We examined the mechanism by which the C-terminal 236 amino acids of the even-skipped protein (region CD) repress transcription. A fusion protein, CDGB, was created that contains region CD fused to the glucocorticoid receptor DNA binding domain. This protein repressed transcription in an in vitro system containing purified fractions of the RNA polymerase II general transcription factors, and repression was dependent upon the presence of high-affinity glucocorticoid receptor binding sites in the promoter. Repression by CDGB was prevented when the promoter DNA was preincubated with TFIID or TBP, whereas preincubation of the template DNA with CDGB prevented TFIID binding. Together, these results strongly imply that CDGB represses transcription by inhibiting TFIID binding, and further experiments suggested a mechanism by which this may occur. Region CD can mediate cooperative interactions between repressor molecules such that molecules bound at the glucocorticoid receptor binding sites stabilize binding of additional CDGB molecules to low-affinity binding sites throughout the basal promoter. Binding to some of these low-affinity sites was shown to contribute to repression. Further experiments suggested that the full-length eve protein also represses transcription by the same mechanism. We speculate that occupancy of secondary sites within the basal promoter by CDGB or the eve protein inhibits subsequent TFIID binding to repress transcription, a mechanism we term cooperative blocking.

Cooperative binding at a distance by even-skipped protein correlates with repression and suggests a mechanism of silencing.

In this study, we examined how the Drosophila developmental control gene even-skipped (eve) represses transcription. Tissue culture cells were used to show that eve contains domains which inhibit transcriptional activators present at the Ultrabithorax (Ubx) proximal promoter when bound up to 1.5 kb away from these activators. Different portions of eve were fused to a heterologous DNA binding domain to show that three adjacent regions of eve contribute to silencing. There appear to be two mechanisms by which eve protein represses transcription. In this study, we used in vitro transcription and DNA binding experiments to provide evidence for one of these mechanisms. Repression in vitro correlates with binding of eve protein to two low-affinity sites in the Ubx proximal promoter. Occupancy of these low-affinity sites is dependent upon cooperative binding of other eve molecules to a separate high-affinity site. Some of these sites are separated by over 150 bp of DNA, and the data suggest that this intervening DNA is bent to form a looped structure similar to those caused by prokaryotic repressors. One of the low-affinity sites overlaps an activator element bound by the zeste transcription factor. Binding of eve protein is shown to exclude binding by zeste protein. These data suggest a mechanism for silencing whereby a repressor protein would be targeted to DNA by a high-affinity element, which itself does not overlap activator elements. Cooperative binding of further repressor molecules to distant low-affinity sites, and competition with activators bound at these sites lead to repression at a distance.

Adrenergic stimulation of diacylglycerol production in genital tract smooth muscle myocytes.

Diacylglycerol (DAG) production has not been reported in previous studies that have characterized inositol phosphate production during alpha-1 adrenergic receptor signal transduction in the DDT1 MF-2 genital tract myocytes. The current study sought to measure norepinephrine (NE)-stimulated DAG production in these transformed myocytes utilizing thin layer chromatography. DAG production was characterized as an alpha-1 adrenergic mediated event utilizing subtype specific adrenergic agonist and antagonists. DAG production occurred in response to physiologic concentration of NE, was apparent by 30 s and was significantly increased by 2 min. Maximal DAG production was unaffected by pretreatment of the myocytes for 96 h with testosterone, which has previously been shown to induce a doubling of alpha-1 adrenergic receptors in these cells. In contrast, testosterone pretreatment did result in a shift of the dose-response curve resulting in a significantly lower EC50 for NE in the treated cells compared to control myocytes. In conclusion, these studies have confirmed that DAG production occurs as a component of alpha-1 adrenergic signal transduction in the DDT1 MF-2 myocytes; transduction events that were modulated by testosterone resulting in increased agonist sensitivity.

Adrenergic stimulation of diacylglycerol production in genital tract smooth muscle myocytes.

Diacylglycerol (DAG) production has not been reported in previous studies that have characterized inositol phosphate production during alpha-1 adrenergic receptor signal transduction in the DDT1 MF-2 genital tract myocytes. The current study sought to measure norepinephrine (NE)-stimulated DAG production in these transformed myocytes utilizing thin layer chromatography. DAG production was characterized as an alpha-1 adrenergic mediated event utilizing subtype specific adrenergic agonist and antagonists. DAG production occurred in response to physiologic concentration of NE, was apparent by 30 s and was significantly increased by 2 min. Maximal DAG production was unaffected by pretreatment of the myocytes for 96 h with testosterone, which has previously been shown to induce a doubling of alpha-1 adrenergic receptors in these cells. In contrast, testosterone pretreatment did result in a shift of the dose-response curve resulting in a significantly lower EC50 for NE in the treated cells compared to control myocytes. In conclusion, these studies have confirmed that DAG production occurs as a component of alpha-1 adrenergic signal transduction in the DDT1 MF-2 myocytes; transduction events that were modulated by testosterone resulting in increased agonist sensitivity.

Asynchronous multiple birth. A report of five cases.

The feasibility of prolonging the delivery interval of the fetus or fetuses in multiple gestations after the preterm delivery of one fetus has been demonstrated. Five clinical reports and a literature review served as the database for this study. Pregnancy was extended in each of five patients with multiple gestations after the extreme preterm delivery of one fetus. Four of the six remaining infants survived. The literature reviewed shows successful survival in 42 of 52 (81%) such asynchronously delivered infants. Use of tocolytic therapy, broad-spectrum antibiotics and cerclage allows pregnancy extension when delivery occurs asynchronously in multiple gestations. The patient's strong desire and full understanding of the potential risks are mandatory before such an endeavor is attempted.

Klossiella equi in the kidneys of a horse.

The protozoan, Klossiella equi was found in the kidneys of an aged Shetland mare raised in the Fredericton area of New Brunswick. This is the first published report of K. equi in a horse in Canada. The microscopic appearance of the parasite in the kidney is described. A brief discussion of other conditions seen in the horse is also presented.

Purification of the Drosophila RNA polymerase II general transcription factors.

We describe a fractionation and purification scheme for the Drosophila RNA polymerase II general transcription factors. Drosophila TFIIE, TFIIF, TFIIH, and RNA polymerase II have been purified to greater than 50% homogeneity from Drosophila embryo nuclear extracts. TFIID has been purified 80-fold and is not significantly contaminated with any of the other general factors. This is the first reported identification and purification of Drosophila TFIIH and TFIIE. Further analysis shows that, similar to their mammalian counterparts, Drosophila TFIIH is composed of eight polypeptides sized between 30 and 100 kDa, and Drosophila TFIIE is composed of two polypeptides sized at 34 and 60 kDa. When all of these fractions are combined with recombinant Drosophila TFlIB, a highly purified in vitro transcription system is generated that has not previously been available in Drosophila. The TFIID fraction can be replaced with recombinant Drosophila TBP to give a transcription system that is nearly free of contaminating proteins.

Coordinate binding of ATP and origin DNA regulates the ATPase activity of the origin recognition complex.

The Origin Recognition Complex (ORC) is a six-protein assembly that specifies the sites of DNA replication initiation in S. cerevisiae. Origin recognition by ORC requires ATP. Here, we demonstrate that two subunits, Orc1p and Orc5p, bind ATP and that Orc1p also hydrolyzes ATP. ATP binding and hydrolysis by Orc1p are both regulated by origin DNA in a sequence-specific manner. ATP binding to Orc1p, but not ATP hydrolysis, is responsible for the ATP dependence of the ORC-origin interaction, indicating that ATP is a cofactor that locks ORC on origin DNA. These data demonstrate that occupancy of the Orc1p ATP-binding site has a profound effect on ORC function and that ATP hydrolysis by Orc1p has the potential to drive transitions between different functional states of ORC.

Suppression of human chorionic gonadotropin in the human placenta at term by human thyroid-stimulating hormone in vitro.

Human chorionic gonadotropin (hCG) exerts a clinically apparent negative feedback on the secretion of human thyroid-stimulating hormone (hTSH) in pregnancy, and the two have cross-reactivity for the TSH receptor in membrane preparations of the thyroid. We examined whether hTSH, in turn, has an influence on the secretion and synthesis of hCG in short-term cultures of human placenta at term. A dose- and time-dependent decrease in the extracellular hCG concentration caused by hTSH was demonstrated. To examine whether hTSH inhibits de novo synthesis of hCG or decreases hCG depletion, we determined the amount of hCG secreted and the size of the intracellular pool by using an enzyme immunoassay. By incorporating a radiolabeled amino acid in the hCG molecule, we measured the amount of hCG synthesized de novo. We concluded that hTSH acts by decreasing the rate of de novo synthesis of placental hCG.

ORC localization in Drosophila follicle cells and the effects of mutations in dE2F and dDP.

We isolated mutations in Drosophila E2F and DP that affect chorion gene amplification and ORC2 localization in the follicle cells. In the follicle cells of the ovary, the ORC2 protein is localized throughout the follicle cell nuclei when they are undergoing polyploid genomic replication, and its levels appear constant in both S and G phases. In contrast, when genomic replication ceases and specific regions amplify, ORC2 is present solely at the amplifying loci. Mutations in the DNA-binding domains of dE2F or dDP reduce amplification, and in these mutants specific localization of ORC2 to amplification loci is lost. Interestingly, a dE2F mutant predicted to lack the carboxy-terminal transcriptional activation and RB-binding domain does not abolish ORC2 localization and shows premature chorion amplification. The effect of the mutations in the heterodimer subunits suggests that E2F controls not only the onset of S phase but also origin activity within S phase.

Drosophila ORC specifically binds to ACE3, an origin of DNA replication control element.

In the yeast Saccharomyces cerevisiae, sequence-specific DNA binding by the origin recognition complex (ORC) is responsible for selecting origins of DNA replication. In metazoans, origin selection is poorly understood and it is unknown whether specific DNA binding by metazoan ORC controls replication. To address this problem, we used in vivo and in vitro approaches to demonstrate that Drosophila ORC (DmORC) binds to replication elements that direct repeated initiation of replication to amplify the Drosophila chorion gene loci in the follicle cells of egg chambers. Using immunolocalization, we observe that ACE3, a 440-bp chorion element that contains information sufficient to drive amplification, directs DmORC localization in follicle cells. Similarly, in vivo cross-linking and chromatin immunoprecipitation assays demonstrate association of DmORC with both ACE3 and two other amplification control elements, AER-d and ACE1. To demonstrate that the in vivo localization of DmORC is related to its DNA-binding properties, we find that purified DmORC binds to ACE3 and AER-d in vitro, and like its S. cerevisiae counterpart, this binding is dependent on ATP. Our findings suggest that sequence-specific DNA binding by ORC regulates initiation of metazoan DNA replication. Furthermore, adaptation of this experimental approach will allow for the identification of additional metazoan ORC DNA-binding sites and potentially origins of replication.

Mometasone furoate is a less specific glucocorticoid than fluticasone propionate.

Fluticasone propionate (FP) and mometasone furoate (MF) are potent synthetic corticosteroids that are widely used as anti-inflammatory agents to treat respiratory diseases. As part of the assessment of the potential for side-effects associated with their use, their activities, not only at the glucocorticoid receptor (GR) but also at the other members of the steroid nuclear receptor family, have been compared. Cell-based functional systems were established to measure different aspects of GR function, as well as the activity at all the other steroid nuclear receptors. The effects of MF and FP on the GR were potent and indistinguishable. Neither corticosteroid showed any activity at the oestrogen receptor, while both were weak antagonists of the androgen receptor. FP was a relatively weak agonist of the progesterone receptor but MF was a very potent agonist of the progesterone receptor, giving activity at similar concentrations to those that stimulate the GR (concentration generating 50% maximal effect (EC50)=50 pM). Moreover, while FP was a weak antagonist of the mineralocorticoid receptor (concentration generating 50% maximal inhibitory effect=80 nM), MF displayed potent partial agonist activity (EC50=3 nM, 30%). Mometasone furoate is considerably less specific for the glucocorticoid receptor than fluticasone propionate, showing significant activity at other nuclear steroid receptors.