Alien Registration- Michaud, Frank (Fort Kent, Aroostook County)

https://digitalmaine.com/alien_docs/36310/thumbnail.jp

BOUND OBJECT HIERARCHY SERVICE

Presented herein is a hierarchical representation of uniquely identified objects within the physical world, referred to as a “Bound Object Hierarchy Service.” The Bound Object Hierarchy Service enables the synchronous shared view and creation of digital twins among coordinating entities. The system enables the representation and policy based access to objects, mapped from the physical world, that would typically be considered to follow a “bound within” hierarchy. The digital custody and ownership of the physical objects can autonomously change over time within the system (e.g., as an object passes along a supply chain, determined via sensors within the environment and policy based access). The policy based approach can additionally enable third parties or a limited set of the system participants to access the object hierarchy data in order to view specific elements of the tracked objects, along with the sharing of object sensor data

A Fatal Overdose of Cocaine Associated with Coingestion of Marijuana, Buprenorphine, and Fluoxetine. Body Fluid and Tissue Distribution of Cocaine and Its Metabolites Determined by Hydrophilic Interaction Chromatography-Mass Spectrometry (HILIC-MS)

Chromatographic separation of highly polar basic drugs with ideal ionspray mass spectrometry volatile mobile phases is a difficult challenge. A new quantification procedure was developed using hydrophilic interaction chromatography-mass spectrometry with turbo-ionspray ionization in the positive mode. After addition of deuterated internal standards and simple clean-up liquid extraction, the dried extracts were reconstituted in 500 μL pure acetonitrile and 5 μL was directly injected onto a Waters Atlantis™ HILIC 150- × 2.1-mm, 3-μm column. Chromatographic separations of cocaine, seven metabolites, and anhydroecgonine were obtained by linear gradient-elution with decreasing high concentrations of acetonitrile (80-56% in 18 min). This high proportion of organic solvent makes it easier to be coupled with MS. The eluent was buffered with 2mM ammonium acetate at pH 4.5. Except for m-hydroxy-benzoylecgonine, the within-day and between-day precisions at 20, 100, and 500 ng/mL were below 7 and 19.1%, respectively. Accuracy was also below ± 13.5% at all tested concentrations. The limit of quantification was 5 ng/mL (%Diff < 16.1, %RSD < 4.3) and the limit of detection below 0.5 ng/mL. This method was successfully applied to a fatal overdose. In Switzerland, cocaine abuse has dramatically increased in the last few years. A 45-year-old man, a known HIV-positive drug user, was found dead at home. According to relatives, cocaine was self-injected about 10 times during the evening before death. A low amount of cocaine (0.45 mg) was detected in the bloody fluid taken from a syringe discovered near the corpse. Besides injection marks, no significant lesions were detected during the forensic autopsy. Toxicological investigations showed high cocaine concentrations in all body fluids and tissues. The peripheral blood concentrations of cocaine, benzoylecgonine, and methylecgonine were 5.0, 10.4, and 4.1 mg/L, respectively. The brain concentrations of cocaine, benzoylecgonine, and methylecgonine were 21.2, 3.8, and 3.3 mg/kg, respectively. The highest concentrations of norcocaine (about 1 mg/L) were measured in bile and urine. Very high levels of cocaine were determined in hair (160 ng/mg), indicating chronic cocaine use. A low concentration of anhydroecgonine methylester was also found in urine (0.65 mg/L) suggesting recent cocaine inhalation. Therapeutic blood concentrations of fluoxetine (0.15 mg/L) and buprenorphine (0.1 μg/L) were also discovered. A relatively high concentration of Δ9-THC was measured both in peripheral blood (8.2 μg/L) and brain cortex (13.5 μg/kg), suggesting that the victim was under the influence of cannabis at the time of death. In addition, fluoxetine might have enhanced the toxic effects of cocaine because of its weak pro-arrhythmogenic properties. Likewise, combination of cannabinoids and cocaine might have increase detrimental cardiovascular effects. Altogether, these results indicate a lethal cocaine overdose with a minor contribution of fluoxetine and cannabinoid

Diagnostic value of tachycardia features and pacing maneuvers during paroxysmal supraventricular tachycardia

AbstractOBJECTIVESThe purpose of this prospective study was to quantitate the diagnostic value of several tachycardia features and pacing maneuvers in patients with paroxysmal supraventricular tachycardia (PSVT) in the electrophysiology laboratory.BACKGROUNDNo study has prospectively compared the value of multiple diagnostic tools in a large group of patients with PSVT.METHODSOne hundred ninety-six consecutive patients who had 200 inducible sustained PSVTs during an electrophysiology procedure were included. The diagnostic values of four baseline electrophysiologic parameters, nine tachycardia features and five diagnostic pacing maneuvers were quantified.RESULTSThe only tachycardia characteristic that was diagnostic of atrioventricular (AV) nodal reentry was a septal ventriculoatrial (VA) time of <70 ms, and no pacing maneuver was diagnostic for AV nodal reentry. An increase in the VA interval with the development of a bundle branch block was the only tachycardia characteristic that was diagnostic for orthodromic tachycardia, but it occurred in only 7% of all tachycardias. An atrial-atrial-ventricular response upon cessation of ventricular overdrive pacing was diagnostic of atrial tachycardia, and this maneuver could be applied to 78% of all tachycardias. Burst ventricular pacing excluded atrial tachycardia when the tachycardia terminated without depolarization of the atrium, but the result could be obtained only in 27% of patients.CONCLUSIONSThis prospective study quantitates the diagnostic value of multiple observations and pacing maneuvers that are commonly used during PSVT in the electrophysiology laboratory. The findings demonstrate that diagnostic techniques rarely provide a diagnosis when used individually. Therefore, careful observations and multiple pacing maneuvers are often required for an accurate diagnosis during PSVT. The results of this study provide a useful reference with which new diagnostic techniques can be compared

Effect of Chronic Amiodarone Therapy on Defibrillation Energy Requirements in Humans

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/73512/1/j.1540-8167.2000.tb00043.x.pd

A chimeric affinity tag for efficient expression and chromatographic purification of heterologous proteins from plants

The use of plants as expression hosts for recombinant proteins is an increasingly attractive option for the production of complex and challenging biopharmaceuticals. Tools are needed at present to marry recent developments in high-yielding gene vectors for heterologous expression with routine protein purification techniques. In this study, we designed the Cysta-tag, a new purification tag for immobilized metal affinity chromatography (IMAC) of plant-made proteins based on the protein-stabilizing fusion partner SlCYS8. We show that the Cysta-tag may be used to readily purify proteins under native conditions, and then be removed enzymatically to isolate the protein of interest. We also show that commonly used protease recognition sites for linking purification tags are differentially stable in leaves of the commonly used expression host Nicotiana benthamiana, with those linkers susceptible to cysteine proteases being less stable then serine protease-cleavable linkers. As an example, we describe a Cysta-tag experimental scheme for the one-step purification of a clinically useful protein, human α1-antitrypsin, transiently expressed in N. benthamiana. With potential applicability to the variety of chromatography formats commercially available for IMAC-based protein purification, the Cysta-tag provides a convenient means for the efficient and cost-effective purification of recombinant proteins from plant tissues.A Discovery grant from the Natural Science and Engineering Research Council (NSERC) of Canada to DM, and by an Australian Research Council (ARC) Discovery Early Career Research Award to FS (DE140101553).PJ was the recipient of an AgroPhytoSciences NSERC–FONCER scholarship and of a BMP graduate scholarship funded by NSERC, the Fonds de Recherche Québec Nature et Technologies and our private research partner Medicago Inc.http://www.frontiersin.orgam2016Plant Production and Soil Scienc

Identification of critical enzymes in the salmon louse chitin synthesis pathway as revealed by RNA interference-mediated abrogation of infectivity

Treatment of infestation by the ectoparasite Lepeophtheirus salmonis relies on a small number of chemotherapeutant treatments that currently meet with limited success. Drugs targeting chitin synthesis have been largely successful against terrestrial parasites where the pathway is well characterised. However, a comparable approach against salmon lice has been, until recently, less successful, likely due to a poor understanding of the chitin synthesis pathway. Post-transcriptional silencing of genes by RNA interference (RNAi) is a powerful method for evaluation of protein function in non-model organisms and has been successfully applied to the salmon louse. In the present study, putative genes coding for enzymes involved in L. salmonis chitin synthesis were characterised after knockdown by RNAi. Nauplii I stage L. salmonis were exposed to double-stranded (ds) RNA specific for several putative non-redundant points in the pathway: glutamine: fructose-6-phosphate aminotransferase (LsGFAT), UDP-N-acetylglucosamine pyrophosphorylase (LsUAP), N-acetylglucosamine phosphate mutase (LsAGM), chitin synthase 1 (LsCHS1), and chitin synthase 2 (LsCHS2). Additionally, we targeted three putative chitin deacetylases (LsCDA4557, 5169 and 5956) by knockdown. Successful knockdown was determined after moulting to the copepodite stage by real-time quantitative PCR (RT-qPCR), while infectivity potential (the number of attached chalimus II compared with the initial number of larvae in the system) was measured after exposure to Atlantic salmon and subsequent development on their host. Compared with controls, infectivity potential was not compromised in dsAGM, dsCHS2, dsCDA4557, or dsCDA5169 groups. In contrast, there was a significant effect in the dsUAP-treated group. However, of most interest was the treatment with dsGFAT, dsCHS1, dsCHS1+2, and dsCDA5956, which resulted in complete abrogation of infectivity, despite apparent compensatory mechanisms in the chitin synthesis pathway as detected by qPCR. There appeared to be a common phenotypic effect in these groups, characterised by significant aberrations in appendage morphology and an inability to swim. Ultrastructurally, dsGFAT showed a significantly distorted procuticle without distinct exo/endocuticle and intermittent electron dense (i.e. chitin) inclusions, and together with dsUAP and dsCHS1, indicated delayed entry to the pre-moult phase.publishedVersio

Electrogram Polarity and Cavotricuspid Isthmus Block During Ablation of Typical Atrial Flutter

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/72217/1/j.1540-8167.2001.00393.x.pd

The temporal reliability of serum estrogens, progesterone, gonadotropins, SHBG and urinary estrogen and progesterone metabolites in premenopausal women

BACKGROUND: There is little existing research to guide researchers in estimating the minimum number of measurement occasions required to obtain reliable estimates of serum estrogens, progesterone, gonadotropins, sex hormone-binding globulin (SHBG), and urinary estrogen and progesterone metabolites in premenopausal women. METHODS: Using data from a longitudinal study of 34 women with a mean age of 42.3 years (SD = 2.6), we calculated the minimum number of measurement occasions required to obtain reliable estimates of 12 analytes (8 in blood, 4 in urine). Five samples were obtained over 1 year: at baseline, and after 1, 3, 6, and 12 months. We also calculated the percent of true variance accounted for by a single measurement and intraclass correlation coefficients (ICC) between measurement occasions. RESULTS: Only 2 of the 12 analytes we examined, SHBG and estrone sulfate (E(1)S), could be adequately estimated by a single measurement using a minimum reliability standard of having the potential to account for 64% of true variance. Other analytes required from 2 to 12 occasions to account for 81% of the true variance, and 2 to 5 occasions to account for 64% of true variance. ICCs ranged from 0.33 for estradiol (E(2)) to 0.88 for SHBG. Percent of true variance accounted for by single measurements ranged from 29% for luteinizing hormone (LH) to 92% for SHBG. CONCLUSIONS: Experimental designs that take the natural variability of these analytes into account by obtaining measurements on a sufficient number of occasions will be rewarded with increased power and accuracy

The effects of phenoxodiol on the cell cycle of prostate cancer cell lines

Background: Prostate cancer is associated with a poor survival rate. The ability of cancer cells to evade apoptosis and exhibit limitless replication potential allows for progression of cancer from a benign to a metastatic phenotype. The aim of this study was to investigate in vitro the effect of the isoflavone phenoxodiol on the expression of cell cycle genes. Methods: Three prostate cancer cell lines-LNCaP, DU145, and PC3 were cultured in vitro, and then treated with phenoxodiol (10 μM and 30 μM) for 24 and 48 h. The expression of cell cycle genes p21WAF1, c-Myc, Cyclin-D1, and Ki-67 was investigated by Real Time PCR. Results: Here we report that phenoxodiol induces cell cycle arrest in the G1/S phase of the cell cycle, with the resultant arrest due to the upregulation of p21WAF1 in all the cell lines in response to treatment, indicating that activation of p21WAF1 and subsequent cell arrest was occurring via a p53 independent manner, with induction of cytotoxicity independent of caspase activation. We found that c-Myc and Cyclin-D1 expression was not consistently altered across all cell lines but Ki-67 signalling expression was decreased in line with the cell cycle arrest. Conclusions: Phenoxodiol demonstrates an ability in prostate cancer cells to induce significant cytotoxicity in cells by interacting with p21WAF1 and inducing cell cycle arrest irrespective of p53 status or caspase pathway interactions. These data indicate that phenoxodiol would be effective as a potential future treatment modality for both hormone sensitive and hormone refractory prostate cancer