Effects of Purified Myosin Light Chain Kinase on Myosin Light Chain Phosphorylation and Catecholamine Secretion in Digitonin-Permeabilized Chromaffin Cells

Many non-muscle cells including chromaffin cells contain actin and myosin. The 20,000 dalton light chain subunits of myosin can be phosphorylated by a Ca 2+ /calmodulin-dependent enzyme, myosin light chain kinase. In tissues other than striated muscle, light chain phosphorylation is required for actin-induced myosin ATPase activity. The possibility that actin and myosin are involved in catecholamine secretion was investigated by determining whether increased phosphorylation in the presence of [γ- 32 P]ATP of myosin light chain by myosin light chain kinase enhances secretion from digitonin-treated chromaffin cells. In the absence of exogenous myosin light chain kinase, 1 μM Ca 2+ caused a 30–40% enhancement of the phosphorylation of a 20 kDa protein. This protein was identified on 2-dimensional gels as myosin light chain by its comigration with purified myosin light chain. Purified myosin light chain kinase (400 μg/ml) in the presence of calmodulin (10 μM) caused little or no enhancement of myosin light chain phosphorylation in the absence of Ca 2+ in digitonin-treated cells. In the presence of 1 μM Ca 2+ , myosin light chain kinase (400 μg/ml) caused an approximately two-fold increase in myosin light chain phosphorylation in digitonin-treated cells in 5 min. The phosphorylation required permeabilization of the cells by digitonin and occurred within the cells rather than in the medium. Myosin light chain kinase-induced phosphorylation of myosin light chain was maximal at 1 μM. Ca 2+ . Under identical conditions to those of the phosphorylation experiments, secretion was unaltered by myosin light chain kinase. The experiments indicate that the phosphorylation of myosin light chain by myosin light chain kinase is not a limiting factor in secretion in digitonin-treated chromaffin cells and suggest that the activation of myosin is not directly involved in secretion from the cells. The experiments also demonstrate the feasibility of investigation of effects of exogenously added proteins on secretion in digitonin-treated cells.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/44193/1/10540_2005_Article_BF01121454.pd

Search for eccentric black hole coalescences during the third observing run of LIGO and Virgo

Despite the growing number of confident binary black hole coalescences observed through gravitational waves so far, the astrophysical origin of these binaries remains uncertain. Orbital eccentricity is one of the clearest tracers of binary formation channels. Identifying binary eccentricity, however, remains challenging due to the limited availability of gravitational waveforms that include effects of eccentricity. Here, we present observational results for a waveform-independent search sensitive to eccentric black hole coalescences, covering the third observing run (O3) of the LIGO and Virgo detectors. We identified no new high-significance candidates beyond those that were already identified with searches focusing on quasi-circular binaries. We determine the sensitivity of our search to high-mass (total mass M>70 M⊙) binaries covering eccentricities up to 0.3 at 15 Hz orbital frequency, and use this to compare model predictions to search results. Assuming all detections are indeed quasi-circular, for our fiducial population model, we place an upper limit for the merger rate density of high-mass binaries with eccentricities 0<e≤0.3 at 0.33 Gpc−3 yr−1 at 90\% confidence level

Ultralight vector dark matter search using data from the KAGRA O3GK run

Among the various candidates for dark matter (DM), ultralight vector DM can be probed by laser interferometric gravitational wave detectors through the measurement of oscillating length changes in the arm cavities. In this context, KAGRA has a unique feature due to differing compositions of its mirrors, enhancing the signal of vector DM in the length change in the auxiliary channels. Here we present the result of a search for U(1)B−L gauge boson DM using the KAGRA data from auxiliary length channels during the first joint observation run together with GEO600. By applying our search pipeline, which takes into account the stochastic nature of ultralight DM, upper bounds on the coupling strength between the U(1)B−L gauge boson and ordinary matter are obtained for a range of DM masses. While our constraints are less stringent than those derived from previous experiments, this study demonstrates the applicability of our method to the lower-mass vector DM search, which is made difficult in this measurement by the short observation time compared to the auto-correlation time scale of DM

The Americleft Project:A Modification of Asher-McDade Method for Rating Nasolabial Esthetics in Patients With Unilateral Cleft Lip and Palate Using Q-sort

In this modern world machine learning and deep learning have become popular choice for analysis and identifying various patterns on data in large volumes. The focus of the thesis work has been on the design of the alternative strategies using machine learning to guide the fuzzer in selecting the most promising test cases. Thesis work mainly focuses on the analysis of the data by using machine learning techniques. A detailed analysis study and work is carried out in multiple phases. First phase is targeted to convert the data into suitable format(pre-processing) so that necessary features can be extracted and fed as input to the unsupervised machine learning algorithms. Machine learning algorithms accepts the input data in form of matrices which represents the dimensionality of the extracted features. Several experiments and run time benchmarks have been conducted to choose most efficient algorithm based on execution time and results accuracy. Finally, the best choice has been implanted to get the desired result. The second phase of the work deals with applying supervised learning over clustering results. The final phase describes how an incremental learning model is built to score the test case logs and return their score in near real time which can act as feedback to guide the fuzzer.I denna moderna värld har maskininlärning och djup inlärning blivit populärt val för analys och identifiering av olika mönster på data i stora volymer. Uppsatsen har fokuserat på utformningen av de alternativa strategierna med maskininlärning för att styra fuzzer i valet av de mest lovande testfallen. Examensarbete fokuserar huvudsakligen på analys av data med hjälp av maskininlärningsteknik. En detaljerad analysstudie och arbete utförs i flera faser. Första fasen är inriktad på att konvertera data till lämpligt format (förbehandling) så att nödvändiga funktioner kan extraheras och matas som inmatning till de oövervakade maskininlärningsalgoritmerna. Maskininlärningsalgoritmer accepterar ingångsdata i form av matriser som representerar dimensionen av de extraherade funktionerna. Flera experiment och körtider har genomförts för att välja den mest effektiva algoritmen baserat på exekveringstid och resultatnoggrannhet. Slutligen har det bästa valet implanterats för att få önskat resultat. Den andra fasen av arbetet handlar om att tillämpa övervakat lärande över klusterresultat. Slutfasen beskriver hur en inkrementell inlärningsmodell är uppbyggd för att få poäng i testfallsloggarna och returnera poängen i nära realtid vilket kan fungera som feedback för att styra fuzzer

Do international human rights treaties improve respect for human rights?

After the non-binding Universal Declaration of Human Rights, many global and regional human rights treaties have been concluded. Critics argue that these are unlikely to have made any actual difference in reality. Others contend that international regimes can improve respect for human rights in state parties, particularly in more democratic countries or countries with a strong civil society devoted to human rights and with transnational links. Our findings suggest that rarely does treaty ratification have unconditional effects on human rights. Instead, improvement in human rights is typically more likely the more democratic the country or the more international non-governmental organizations its citizens participate in. Conversely, in very autocratic regimes with weak civil society, ratification can be expected to have no effect and is sometimes even associated with more rights violation