Softmax

Fluorimeter /Max from Molecular Devices (Sunnyvale, CA), SoftMax analysis software (see Note 5). 

Microtiter plate readers were developed for ELISA techniques and this is reflected in the computer software available. The Molecular Devices reader is recommended since the software (Softmax ) was developed for diverse applications and contains a curve fitting facility that is ideal for analysis of survival curves. 

Analyses were done with a Molecular Devices microtiterplate reader interfaced to an IBM computer with Softmax software(Molecular Devices). Either the rate of color change (mOD/min) at 450 nm was recorded using the kinetic format or the final optical density (OD) was measured by difference of readings at 490 and 650 nm. 

Apparatus. Quantitation of color developed in the microwells was achieved by a Vmax Reader (Molecular Devices Corp., Palo Alto, California). Standard curve using semi-log curve fit and unknown calculations were accomplished by using the software program Softmax provided by the manufacturer. The Ultrawasher II (Dynatech Laboratories Alexandria, Virginia) was used to wash the wells. 

The final output is calculated by a softmax function, controlled by the gain parameter G, over all the units in the output layer ... 

The network consists of two input neurons for presentation of the two v-values as well as four output neurons, y, which represent the four classes (cf Figure 8.15). In addition, a hidden layer was added with up to 20 neurons and the intercepts of the surfaces are modeled by bias neurons to both the hidden and output layers. The transfer function in the neurons of the hidden layer was of sigmoid type, and aggregation of the neurons in the output layer was carried out by calculating the normalized exponentials (softmax criterion). 

By assigning a softmax activation function on the output layer of the neural network (or a softmax component in a component-based neural network) for categorical target variables, the outputs can be interpreted as posterior probabilities. This is very useful in classification as it gives a certainty measure on classifications. 

Determination of AGEs is based on the classic spectrofluorimetric detection 18-19. Fluorescence intensity was recorded at the emission maximum 440 nm upon excitation at 370 nm and at emission maximum 385 nm upon excitation at 335 nm for pentosidine fluorescence Fluorescence intensity is expressed in arbitrary units (AU) and corrected for dilutions. We employ a SPECTRAmax Gemini XPS spectrofluorometer with SOFTmax PRO software (Molecular Devices, Sunnyvale, CA). 

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