MOTIVATION: Real time quantitative polymerase chain reaction (qPCR) is an important tool in quantitative studies of DNA and RNA molecules; especially in transcriptome studies, where different primer combinations allow identification of specific transcripts such as splice variants or precursor messenger RNA. Several softwares that implement various rules for optimal primer design are available. Nevertheless, as designing qPCR primers needs to be done manually, the repeated task is tedious, time consuming and prone to errors. RESULTS: We used a set of rules to automatically design all possible exon-exon and intron-exon junctions in the human and mouse transcriptomes. The resulting database is included as a track in the UCSC genome browser, making it widely accessible and easy to use. AVAILABILITY: The database is available from the UCSC genome browser (http://genome.ucsc.edu/), track name 'Whole Transcriptome qPCR Primers' for the hg19 (Human) and mm10 (Mouse) genome versions. Batch query is available in the following: http://www.weizmann.ac.il/complex/compphys/software/Amit/primers/batch_query_qpcr_primers.htm CONTACT: [email protected] or [email protected] SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

BACKGROUND: In many microarray experiments, analysis is severely hindered by a major difficulty: the small number of samples for which expression data has been measured. When one searches for differentially expressed genes, the small number of samples gives rise to an inaccurate estimation of the experimental noise. This, in turn, leads to loss of statistical power. RESULTS: We show that the measurement noise of genes with similar expression levels (intensity) is identically and independently distributed, and that this (intensity dependent) distribution is approximately normal. Our method can be easily adapted and used to test whether these statement hold for data from any particular microarray experiment. We propose a method that provides an accurate estimation of the intensity-dependent variance of the noise distribution, and demonstrate that using this estimation we can detect differential expression with much better statistical power than that of standard t-test, and can compare the noise levels of different experiments and platforms. CONCLUSIONS: When the number of samples is small, the simple method we propose improves significantly the statistical power in identifying differentially expressed genes.

MOTIVATION: Existing computational methods that identify transcription factor (TF) binding sites on a gene's promoter are plagued by significant inaccuracies. Binding of a TF to a particular sequence is assessed by comparing its similarity score, obtained from the TF's known position weight matrix (PWM), to a threshold. If the similarity score is above the threshold, the sequence is considered a putative binding site. Determining this threshold is a central part of the problem, for which no satisfactory biologically based solution exists. RESULTS: We present here a method that integrates gene expression data with sequence-based scoring of TF binding sites, for determining a global score threshold for each TF. We validate our method, STOP (Searching TFs Of Promoters), in several ways: (1) we calculate the average expression values of groups of human putative target genes of each TF, and compare them to similar averages derived for random gene groups. The groups of putative targets show significantly higher relative average expression. (2) We find high consistency between the induced lists of putative targets in human and in mouse. (3) The expression patterns associated with human and mouse genes (ordered by PWM scores for each TF) exhibit high similarity between human and mouse, indicating that our method has firm biological basis. (4) Comparison of results obtained by STOP and PRIMA (Elkon et al., 2003) suggests that determining the score threshold using gene expression, as is done in STOP, is more biologically tuned. AVAILABILITY: Software package will be available for academic users upon request. SUPPLEMENTARY INFORMATION: Supplementary data are available on Bioinformatics online.

Finding the entropy rate of Hidden Markov Processes is an active research topic, of both theoretical and practical importance. A recently used approach is studying the asymptotic behavior of the entropy rate in various regimes. In this paper we generalize and prove a previous conjecture relating the entropy rate to entropies of finite systems. We use the proof to establish series expansions for the entropy rate in two different regimes. We also study the radius of convergence of the two series expansions.

A recent result presented the expansion for the entropy rate of a hidden Markov process (HMP) as a power series in the noise variable epsi. The coefficients of the expansion around the noiseless (epsi=0) limit were calculated up to 11th order, using a conjecture that relates the entropy rate of an HMP to the entropy of a process of finite length (which is calculated analytically). In this letter, we generalize and prove the conjecture and discuss its theoretical and practical consequences

We calculate the Shannon entropy rate of a binary hidden Markov process (HMP), of given transition rate and noise /spl epsiv/ (emission), as a series expansion in /spl epsiv/. The first two orders are calculated exactly. We then evaluate, for finite histories, simple upper-bounds of Cover and Thomas. Surprisingly, we find that for a fixed order k and history of n steps, the bounds become independent of n for large enough n. This observation is the basis of a conjecture, that the upper-bound obtained for n/spl ges/(k+3)/2 gives the exact entropy rate for any desired order k of /spl epsiv/.

MOTIVATION: Predicting the metastatic potential of primary malignant tissues has direct bearing on the choice of therapy. Several microarray studies yielded gene sets whose expression profiles successfully predicted survival. Nevertheless, the overlap between these gene sets is almost zero. Such small overlaps were observed also in other complex diseases, and the variables that could account for the differences had evoked a wide interest. One of the main open questions in this context is whether the disparity can be attributed only to trivial reasons such as different technologies, different patients and different types of analyses. RESULTS: To answer this question, we concentrated on a single breast cancer dataset, and analyzed it by a single method, the one which was used by van't Veer et al. to produce a set of outcome-predictive genes. We showed that, in fact, the resulting set of genes is not unique; it is strongly influenced by the subset of patients used for gene selection. Many equally predictive lists could have been produced from the same analysis. Three main properties of the data explain this sensitivity: (1) many genes are correlated with survival; (2) the differences between these correlations are small; (3) the correlations fluctuate strongly when measured over different subsets of patients. A possible biological explanation for these properties is discussed. CONTACT: [email protected] SUPPLEMENTARY INFORMATION: http://www.weizmann.ac.il/physics/complex/compphys/downloads/liate/

SUMMARY: We introduce a novel unsupervised approach for the organization and visualization of multidimensional data. At the heart of the method is a presentation of the full pairwise distance matrix of the data points, viewed in pseudocolor. The ordering of points is iteratively permuted in search of a linear ordering, which can be used to study embedded shapes. Several examples indicate how the shapes of certain structures in the data (elongated, circular and compact) manifest themselves visually in our permuted distance matrix. It is important to identify the elongated objects since they are often associated with a set of hidden variables, underlying continuous variation in the data. The problem of determining an optimal linear ordering is shown to be NP-Complete, and therefore an iterative search algorithm with O(n3) step-complexity is suggested. By using sorting points into neighborhoods, i.e. SPIN to analyze colon cancer expression data we were able to address the serious problem of sample heterogeneity, which hinders identification of metastasis related genes in our data. Our methodology brings to light the continuous variation of heterogeneity--starting with homogeneous tumor samples and gradually increasing the amount of another tissue. Ordering the samples according to their degree of contamination by unrelated tissue allows the separation of genes associated with irrelevant contamination from those related to cancer progression. AVAILABILITY: Software package will be available for academic users upon request.

MOTIVATION: Genes are often characterized dichotomously as either housekeeping or single-tissue specific. We conjectured that crucial functional information resides in genes with midrange profiles of expression. RESULTS: To obtain such novel information genome-wide, we have determined the mRNA expression levels for one of the largest hitherto analyzed set of 62 839 probesets in 12 representative normal human tissues. Indeed, when using a newly defined graded tissue specificity index tau, valued between 0 for housekeeping genes and 1 for tissue-specific genes, genes with midrange profiles having 0.15< tau<0.85 were found to constitute >50% of all expression patterns. We developed a binary classification, indicating for every gene the I(B) tissues in which it is overly expressed, and the 12-I(B) tissues in which it shows low expression. The 85 dominant midrange patterns with I(B)=2-11 were found to be bimodally distributed, and to contribute most significantly to the definition of tissue specification dendrograms. Our analyses provide a novel route to infer expression profiles for presumed ancestral nodes in the tissue dendrogram. Such definition has uncovered an unsuspected correlation, whereby de novo enhancement and diminution of gene expression go hand in hand. These findings highlight the importance of gene suppression events, with implications to the course of tissue specification in ontogeny and phylogeny. AVAILABILITY: All data and analyses are publically available at the GeneNote website, http://genecards.weizmann.ac.il/genenote/ and, GEO accession GSE803. CONTACT: [email protected] SUPPLEMENTARY INFORMATION: Four tables available at the above site.

UNLABELLED: The F2CS server provides access to the software, F2CS2.00, which implements an automated prediction method of SCOP and CATH classifications of proteins, based on their FSSP Z-scores. AVAILABILITY: Free at http://www.weizmann.ac.il/physics/complex/compphys/f2cs/ SUPPLEMENTARY INFORMATION: The site contains links to additional figures and tables.

Abstract Summary: The CTWC server provides access to the software, CTWC1.00, that implements Coupled Two Way Clustering (Getz et al., 2000), a method designed to mine gene expression data Availability: Free, at http://ctwc.weizmann.ac.il Contact: [email protected] Supplementary information: The site has a link to an example which provides figures and detailed explanations * To whom correspondence should be addressed.

UNLABELLED: We present and review coupled two-way clustering, a method designed to mine gene expression data. The method identifies submatrices of the total expression matrix, whose clustering analysis reveals partitions of samples (and genes) into biologically relevant classes. We demonstrate, on data from colon and breast cancer, that we are able to identify partitions that elude standard clustering analysis. AVAILABILITY: Free, at http://ctwc.weizmann.ac.il.. SUPPLEMENTARY INFORMATION: http://www.weizmann.ac.il/physics/complex/compphys/bioinfo2/

We introduce and study an artificial neural network inspired by the probabilistic receptor affinity distribution model of olfaction. Our system consists of N sensory neurons whose outputs converge on a single processing linear threshold element. The system's aim is to model discrimination of a single target odorant from a large number p of background odorants within a range of odorant concentrations. We show that this is possible provided p does not exceed a critical value p(c) and calculate the critical capacity alpha(c) = p(c)/N. The critical capacity depends on the range of concentrations in which the discrimination is to be accomplished. If the olfactory bulb may be thought of as a collection of such processing elements, each responsible for the discrimination of a single odorant, our study provides a quantitative analysis of the potential computational properties of the olfactory bulb. The mathematical formulation of the problem we consider is one of determining the capacity for linear separability of continuous curves, embedded in a large-dimensional space. This is accomplished here by a numerical study, using a method that signals whether the discrimination task is realizable, together with a finite-size scaling analysis.

We introduce a method for validation of results obtained by clustering analysis of data. The method is based on resampling the available data. A figure of merit that measures the stability of clustering solutions against resampling is introduced. Clusters that are stable against resampling give rise to local maxima of this figure of merit. This is presented first for a one-dimensional data set, for which an analytic approximation for the figure of merit is derived and compared with numerical measurements. Next, the applicability of the method is demonstrated for higher-dimensional data, including gene microarray expression data.

We present a new approach to clustering, based on the physical properties of an inhomogeneous ferromagnet. No assumption is made regarding the underlying distribution of the data. We assign a Potts spin to each data point and introduce an interaction between neighboring points, whose strength is a decreasing function of the distance between the neighbors. This magnetic system exhibits three phases. At very low temperatures, it is completely ordered; all spins are aligned. At very high temperatures, the system does not exhibit any ordering, and in an intermediate regime, clusters of relatively strongly coupled spins become ordered, whereas different clusters remain uncorrelated. This intermediate phase is identified by a jump in the order parameters. The spin-spin correlation function is used to partition the spins and the corresponding data points into clusters. We demonstrate on three synthetic and three real data sets how the method works. Detailed comparison to the performance of other techniques clearly indicates the relative success of our method.

We consider two-layered perceptions consisting of N binary input units, K binary hidden units and one binary output unit, in the limit N >> K > or = 1. We prove that the weights of a regular irreducible network are uniquely determined by its input-output map up to some obvious global symmetries. A network is regular if its K weight vectors from the input layer to the K hidden units are linearly independent. A (single layered) perceptron is said to be irreducible if its output depends on every one of its input units; and a two-layered perceptron is irreducible if the K + 1 perceptrons that constitute such network are irreducible. By global symmetries we mean, for instance, permuting the labels of the hidden units. Hence, two irreducible regular two-layered perceptrons that implement the same Boolean function must have the same number of hidden units, and must be composed of equivalent perceptrons.

A classification problem in high energy physics has been solved on simulated data using a simple multilayer perceptron comprising binary units which was trained with the CHIR algorithm. The unstable training of such a network on a nonseparable set has been overcome by selecting those weight vectors with good performance while providing a flexible choice of the two types of classification errors. Specific features of the problem have been exploited in order to simplify and optimize the solution which has been compared to the popular backpropagation algorithm and found to perform on a similar level. Additional aspects of this work are the use of the CHIR algorithm on continuous input and incorporating the classic idea of a phi-machine in a multilayer perceptron.

The implementation of an all-optical Hopfield-type neural network is made possible by eliminating the need for subtracting light intensities. This implementation is achieved without significantly degrading the network's performance, when only inhibitory connections (i.e., Jij <0) are used. The theoretical analysis of such a network, and its experimental implementation, exploiting a liquid crystal light valve for the neurons and an array of subholograms for the interconnections, are presented.

We present a local perceptron-learning rule that either converges to a solution, or establishes linear nonseparability. We prove that when no solution exists, the algorithm detects this in a finite time (number of learning steps). This time is polynomial in typical cases and exponential in the worst case, when the set of patterns is nonstrictly linearly separable. The algorithm is local and has no arbitrary parameters.