Directed protein evolution provides a series of fluorescent protein-based indicators for multicolor Ca2
+
imaging.
Engineered fluorescent protein (FP) chimeras that modulate their fluorescence in ...response to changes in calcium ion (Ca
2+
) concentration are powerful tools for visualizing intracellular signaling activity. However, despite a decade of availability, the palette of single FP-based Ca
2+
indicators has remained limited to a single green hue. We have expanded this palette by developing blue, improved green, and red intensiometric indicators, as well as an emission ratiometric indicator with an 11,000% ratio change. This series enables improved single-color Ca
2+
imaging in neurons and transgenic
Caenorhabditis elegans
. In HeLa cells, Ca
2+
was imaged in three subcellular compartments, and, in conjunction with a cyan FP–yellow FP–based indicator, Ca
2+
and adenosine 5′-triphosphate were simultaneously imaged. This palette of indicators paints the way to a colorful new era of Ca
2+
imaging.
A main challenge in deformation estimation for fluorescent imaging is to decrease the effects of unavoidable random photon shot noise of fluorescence. To solve this open problem, an efficient ...second-order minimization (ESM) based non-rigid deformation estimation method on fluorescent imaging of neurons is proposed as a visual aid tool for understanding the relationship of neuron activities and behaviors. Because local features, such as corners, lines, arc segments, usually used for deformation estimation, could be compromised by fluorescence noise, global intensity information of all the pixels in a region of interest (ROI) is used as a texture pattern to guide parameterized deformation estimation. By satisfying this principle, three deformation models including affine, homography and Thin Plate Spline (TPS) based on the ESM algorithm are implemented and evaluated. The experimental results illustrate that the homography is the best choice of correctly and robustly estimating parameters for fluorescent deformations under our experimental condition. By using these parameters, the fluorescent intensity in restored image can be measured and analyzed without the disturbance of noise and deformation which can be significant for understanding how nervous system affects and controls animals' behaviors.
Tropomyosin (TM) is one of the actin-regulatory proteins and stabilizes the filamentous structure of F-actin. Although it has been suggested that multiple TM isoforms play important roles in ...nonmuscle cells and several TM-binding proteins have been identified and characterized, the functional properties of nonmuscle TM isoforms are not well understood. In order to investigate the roles of TM in nonmuscle cells, we searched for novel nonmuscle TM-binding proteins from the chicken liver total acetone powder extract by TM-affinity column chromatography. As a consequence, a protein of 57 kDa was mainly eluted from the column and its partial amino acid sequence of the protease-digested fragment was determined. Homology search using SWISS-PLOT showed that the 57 kDa protein was nearly identical to chicken glutamate dehydrogenase (GDH: EC 1.4.1.3). Purified GDH had the ability to bind to the TM-affinity column, indicating that GDH associated directly with liver TM. The binding of GDH to TM was abolished in the presence of 0.1 mM ATP, that is known to decrease the rate of the GDH enzymatic activity at pH 7.5. Thus, we have demonstrated the interaction between nonmuscle TM and GDH for the first time.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
In this paper, we present and construct a distributed problem solving environment (PSE) for partial differential equation (PDE) based problems. The system inputs a problem information including a ...discretization and computation scheme, and outputs a program flow and also a C-language source code for the problem. On a host computer a user inputs his/her problem, and the host navigates the user to solve the problem. The distributed PSE for PDEs consists of several modules: a problem description module, a discretization one, an equation manipulation one, a program design one and a program generation one. Each module is distributed on distributed computers, and the whole information is described by the Extensible Markup Language (XML) including the Mathematical Markup Language (MathML); each distributed module communicates with the host module by using XML documents, so that outputs from each modules are visualized. Independent modules which are developed by other engineers or users for one of the functions specified above can be also used after adjustments to the distributed PSE interface, if necessary. Therefore the concept of the distributed PSE extends the potential of conventional PSE systems. The system is implemented by the Java language with the Document Object Model (DOM), in order to make the system a networked and distributed PSE.
Tropomyosin (TM) is one of the actin-regulatory proteins and stabilizes the filamentous structure of F-actin. Although it has been suggested that multiple TM isoforms play important roles in ...nonmuscle cells and several TM-binding proteins have been identified and characterized, the functional properties of nonmuscle TM isoforms are not well understood. In order to investigate the roles of TM in nonmuscle cells, we searched for novel nonmuscle TM-binding proteins from the chicken liver total acetone powder extract by TM-affinity column chromatography. As a consequence, a protein of 57 kDa was mainly eluted from the column and its partial amino acid sequence of the protease-digested fragment was determined. Homology search using SWISS-PLOT showed that the 57 kDa protein was nearly identical to chicken glutamate dehydrogenase (GDH: EC 1.4.1.3). Purified GDH had the ability to bind to the TM-affinity column, indicating that GDH associated directly with liver TM. The binding of GDH to TM was abolished in the presence of 0.1 mM ATP, that is known to decrease the rate of the GDH enzymatic activity at pH 7.5. Thus, we have demonstrated the interaction between nonmuscle TM and GDH for the first time.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
There is a difference in the vibration characteristics between the real building and the analysis model used at the design stage. In this research, the micro-tremor measurement, the free vibration ...test by the human power and the seismic observation were carried out for the steel structure office building of 14 stories. These tests and observation results were examined from the small vibration level to the medium-earthquake response revel. And the validity of the dynamic analysis model used at the design stage was discussed.
A new type of high-damping rubber damper for vibration control of tall buildings has been being developed in this study. This paper describes the temperature dependency of the damper which will be an ...important problem in vibration control systems using high molecule material. The tests were carried out in two different conditions from the viewpoint of the temperature variation of vibration control material into the dampers. One is the variation of environmental temperature around the damper and the other is the self-generated heat in the damper which will be induced by the cyclic loading such as earthquake or wind. Moreover, the durability of damper was investigated by following tests; breaking test, static tests and cyclic loading tests under excessive shear strain amplitude. As the results, it was confirmed that the temperature dependency of the damper would not much affect to the energy absorbing characteristics even in the case of the practical use and the durability of the damper was highly enough against the large earthquakes by means of its strong adhesive characteristics.
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