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Deconvolution of concentration recordings at live cell preparations via shape error optimization.

In many fields of science and engineering including several areas in analytical chemistry, deconvolution needs to be performed on measured data to extract meaningful information. This situation arises when a variable of interest has to be indirectly estimated from a measurable quantity that depends on this variable in some known manner. This dependence, called the "forward problem", has to be computationally undone to obtain the information sought from the experimental results. Solving this "inverse problem" requires deconvolution whenever the forward problem involves convolution. Despite its ubiquitous importance, however, performance of the methodologies used for deconvolution remains often unsatisfactory. An example is in bioanalytical applications where microsensing at live preparations is performed to obtain information on biological transport. It is in this context that a novel approach to solve inverse problems, shape error optimization, is proposed and tested in this work. The experimental paradigm addressed is in the area of multidrug resistance (MDR) in cancer that gives rise to passive and active drug efflux from cells. Doxorubicin (DOX) concentration is monitored with a carbon fiber microelectrode in vitro at close proximity to a monolayer of cells expressing MDR. The measured local concentration is the result of convolution of cellular efflux with the impulse response of diffusion in the extracellular medium. Hence, estimating DOX efflux, which is the biologically meaningful information, leads to a deconvolution problem. Performance of deconvolution via shape error optimization is compared with that of two conventional techniques: discrete Fourier transform and square error optimization. The results obtained are also applicable to other areas of science and engineering where deconvolution is commonly used for processing experimental data.

Algorithms↗

Validation of deconvolution analysis of LH secretion and half-life.

Deconvolution methods constitute a class of analytic tools to quantitate hormone secretion and/or clearance in vivo. Although mathematically rigorous, deconvolution techniques have assumed, rather than proven, validity. Accordingly, we tested the validity of deconvolution analysis on true-positive human, animal (sheep and monkey), and computer-simulated data using the luteinizing hormone (LH) pulse signal as a relevant paradigm. We found that multiparameter deconvolution analysis has high discriminative sensitivity (human data 91%, animal 81%, computer-stimulated 95%) and specificity (human 90%, animal 81%, computer-simulated 100%). Sensitivity was impaired by low secretory burst amplitude (< 0.1 IU.l-1.min-1), short interpulse interval (< 60 min), infrequent venous sampling (every 20-30 min), and high random experimental variation (e.g., noise > 15%). Specificity was hindered by noise. Deconvolution accurately characterized the unknown hormone half-life (r = +0.994) and production rate (r = +0.990). Interoperator reliability was high when statistically based criteria for secretory pulse detection were applied. We conclude that multiparameter deconvolution, within recognizable constraints, is a valid and reliable tool for in vivo investigation of hormone secretion and half-life.

Algorithms↗

Improving the repeatability of topographic height measurements in confocal scanning laser imaging using maximum-likelihood deconvolution.

PURPOSE: To evaluate maximum likelihood (ML) blind deconvolution as a technique for improving the repeatability of topographic height measurements obtained from scanning laser tomography (Heidelberg Retinal Tomograph [HRT]; Heidelberg Engineering, Heidelberg, Germany). METHODS: ML blind deconvolution is an image-processing technique that estimates the original scene from a degraded image. This technique has been used in confocal scanning laser microscopy to remove "out-of-focus" haze in three-dimensional confocal image stacks. ML blind deconvolution requires no prior estimation of the point-spread function (PSF), as opposed to classic linear deconvolution methods. Instead, the algorithm estimates an initial PSF based on the optical setup of the confocal scanning device and optics of the eye and iteratively proceeds to a solution. The improvement in repeatability of height measurements from mean topography images within scan (intrascan) and between scans (interscan) afforded by ML deconvolution was evaluated in a test-retest series of HRT images from 40 ocular hypertensive and glaucomatous patients with varying degrees of media opacity. RESULTS: There was an improvement in intrascan repeatability in 38 out of the 40 mean topography images (median improvement 2.5 microm, inter-quartile range 2.19, P < 0.001), and an improvement in interscan repeatability in 33 of the 40 mean topographies (median improvement, 1.0 microm, interquartile range 3.49, P < 0.001). There was a positive association between the magnitude of the improvement in repeatability and the level of mean pixel height standard deviation (MPHSD), intrascan (P = 0.004) and interscan (P = 0.002). CONCLUSIONS: ML blind deconvolution algorithm improves the repeatability of topographic height measurements from the HRT. This improvement was greater in patients with poorer quality images.

Algorithms↗

The use of a physiologically based pharmacokinetic model to evaluate deconvolution measurements of systemic absorption.

BACKGROUND: An unknown input function can be determined by deconvolution using the systemic bolus input function (r) determined using an experimental input of duration ranging from a few seconds to many minutes. The quantitative relation between the duration of the input and the accuracy of r is unknown. Although a large number of deconvolution procedures have been described, these routines are not available in a convenient software package. METHODS: Four deconvolution methods are implemented in a new, user-friendly software program (PKQuest, http://www.pkquest.com). Three of these methods are characterized by input parameters that are adjusted by the user to provide the "best" fit. A new approach is used to determine these parameters, based on the assumption that the input can be approximated by a gamma distribution. Deconvolution methodologies are evaluated using data generated from a physiologically based pharmacokinetic model (PBPK). RESULTS AND CONCLUSIONS: The 11-compartment PBPK model is accurately described by either a 2 or 3-exponential function, depending on whether or not there is significant tissue binding. For an accurate estimate of r the first venous sample should be at or before the end of the constant infusion and a long (10 minute) constant infusion is preferable to a bolus injection. For noisy data, a gamma distribution deconvolution provides the best result if the input has the form of a gamma distribution. For other input functions, good results are obtained using deconvolution methods based on modeling the input with either a B-spline or uniform dense set of time points.

Absorption↗

In vivo dynamics of luteinizing hormone secretion and clearance in man: assessment by deconvolution mechanics.

The dynamics of LH secretion and clearance were studied in vivo in eight healthy young men using a novel multiple parameter deconvolution procedure to resolve underlying secretion and clearance rates simultaneously from serial serum immunoactive LH concentrations. This deconvolution analysis disclosed random LH secretory bursts occurring at a mean (+/- SEM) interpulse interval of 72 +/- 5 min. The frequency of these secretory bursts was 6.9 +/- 0.6 episodes/8 h. Each resolved LH secretory event had an average half-duration of only 7.8 +/- 0.5 min, which was remarkably shorter than the LH concentration peak duration of 60 +/- 6 min. The maximal LH secretory rate achieved within a secretory burst averaged 0.40 +/- 0.05 mIU/min.mL (0.14 IU/min.L), which corresponded to a mass of LH released of 3.2 +/- 0.3 mIU/mL distribution vol (IU/L). By linear regression analysis, both the maximal rate and mass of LH released per secretory burst were positively correlated with the duration of the subsequent interpulse interval (P less than 0.001). In physiological experiments, the mass of LH released per secretory burst was increased by iv GnRH injections or primary gonadal failure, and decreased by sc administration of a selective GnRH antagonist (Nal,Glu-GnRH). The mean endogenous LH production rate calculated by deconvolution [180 +/- 40 mIU/min (0.18 IU/min)] was not different from a nominal value of 228 +/- 80 mIU/min (0.228 IU/min) extrapolated from earlier steady state LH infusions. Deconvolution estimated single phase half-times of endogenous LH disappearance of 87 +/- 8 min, were in general harmony with values of 44-106 min obtained previously in four LH-deficient men injected with purified LH. Moreover, creation of synthetic LH series using our deconvolution estimates yielded 24-h LH pulse profiles similar quantitatively and qualitatively to those in normal men. In summary, we applied a new multiple parameter deconvolution procedure to immunoactive LH pulse profiles to discern the nature of physiological LH secretory events and estimate endogenous LH disappearance rates. These studies in normal men have revealed that 1) LH secretory bursts occur every 72 min; their mean half-duration is 7.8 min; and 3.2 mIU/mL (3.2 IU/L) are released per secretory peak; 2) amplitudes of in vivo LH secretory bursts are augmented in primary gonadal failure and/or by exogenous GnRH injections and attenuated by a GnRH antagonist; 3) the duration of a postsecretory pause is proportional to the amplitude of the preceding LH secretory burst; and (4) 95% of total daily LH release occurs in only 5 h/day.(ABSTRACT TRUNCATED AT 400 WORDS)

Adult↗

Five modified numerical deconvolution methods for biopharmaceutics and pharmacokinetics studies.

Four improved finite-difference numerical deconvolution methods and one nonlinear regression numerical deconvolution method are proposed and implemented using IMSL/IDLTM. These five numerical deconvolution methods are evaluated using simulated data generated with and without added noise under six different dosing cases. Comparisons between these methods are made in terms of the superimposability of the calculated cumulative amount of drug released or absorbed-time profiles with the theoretical data. The results indicate that the proposed fixed step number equal step length numerical deconvolution method is simple and accurate and therefore is appropriate for pharmacokinetic and biopharmaceutic studies. When an analytic function is legitimate to represent the drug input rate, the nonlinear regression numerical deconvolution method will yield enhanced numerical accuracy and stability.

Absorption↗

Comparison of three deconvolution techniques in renography.

The importance of comparing various deconvolution techniques on real patient data lies in the fact that there are two different approaches to that problem. One, which prefers general-purpose techniques, using a matrix algorithm (MA) and fast-Fourier transformation (FFT) with various smoothings, filters, etc., and another which prefers deconvolution techniques which have been modified for the particular application. The modification may be in requiring well defined input or assuming a smooth result of deconvolution. The aim of this paper is to compare the general-purpose deconvolution technique mostly used in analyzing patient data in nuclear medicine using MA and FFT with deconvolution based on Laplace transformation (LT). They have been compared on real patient data renograms in 36 cases. The comparison shows that although the LT technique requires a well defined input and for that reason needs more computing time than the FFT and MA technique, it gives impulse response functions (IRF) without negative values or with negligible ones. Even more, using LT, which has been incorporated in a model of distribution and transport of tracer, without taking blood samples one can assess some other useful clinical parameters such as inverse value of clearance time constant (ICTC) and effective renal plasma flow (ERPF).

Humans↗

Quantification of left to right cardiac shunts by multiple deconvolution analysis.

A new method for quantification of left to right cardiac shunts was studied in 17 patients scheduled for cardiac catheterization who had also undergone radionuclide angiocardiography. The observed pulmonary transit curve was deconvoluted in two different ways: (1) by the superior vena caval ("bolus") time-activity curve, to yield the deconvoluted pulmonary transit curve, which represented the theoretical pulmonary transit curve with a perfect bolus injection, and (2) by the right ventricular time-activity curve, to yield the pulmonary transfer function, which represented the theoretical pulmonary transit curve with a perfect bolus injection an with no intracardiac shunts. The pulmonary transfer function was superimposed on the deconvoluted pulmonary transit curve, and the area A under it obtained. The pulmonary transfer function was then subtracted from the deconvoluted pulmonary transit curve. The pulmonary transfer function was scaled to fit the resulting shunt recirculation peak in the difference curve, and the area B under this scaled pulmonary transfer function obtained. Shunt size was quantified as the pulmonary (QP) to systemic (QS) flow ration QP/QS = A/(A-B). The method correlated closely with oximetry (r = 0.93). Use of this multiple deconvolution analysis technique provides accurate shunt quantification and reduces subjective operator decisions.

Adolescent↗

Deconvolution analysis of hormone data.

Deconvolution analysis of hormone data poses special problems in view of the sparse, noisy, and short data series typically available for analysis; the unknown true nature of the underlying secretory event; and potentially large variations in dissipation or clearance kinetics in different settings. Consequently, deconvolution techniques, which concern themselves with the estimation of hormone secretion and/or clearance based on serial circulating hormone concentration measurements, face a particular challenge. Ideal features of deconvolution algorithms are summarized in Table IV. Specific deconvolution techniques available to analyze hormone data include both waveform-defined procedures and waveform-independent algorithms. These approaches should be viewed as complementary rather than antagonistic. All deconvolution techniques are subject to individual limitations and specific strengths. Independently of the method employed, error propagation is necessary so as to define the statistical uncertainty intrinsic to the estimate of secretion and clearance. Such calculations of experimental uncertainty should include error inherent in the sample collection, processing, and assay as well as error in the kinetic constants and/or anticipated departures of the biological process from the algebraic structure of the convolution formulation. Moreover, more complex convolution statements will be required to describe the full range of behavior of hormone data in a systems view. The applications of such newer convolution methods as well as currently available techniques include model synthesis, model testing, and analysis of the interactions among multiple pulse generators.

Algorithms↗

Automatic program for peak detection and deconvolution of multi-overlapped chromatographic signals part I: peak detection.

A series of two papers describing a procedure for automated peak deconvolution is presented. The goal is to develop a package of routines that can be used by non-experienced users. Part I (this paper) concerns peak detection, whereas Part II is dedicated to the deconvolution itself. In this first part, the most interesting features of the peak detection algorithms, which precede the deconvolution step, are outlined. High-order derivatives provide valuable information to assess the number of underlying compounds under a given peak cluster. A smoothing technique was found essential to compute properly the derivatives, since the noise is amplified when differences are calculated. The Savitsky-Golay smoother was applied in combination with the Durbin-Watson criterion to automate the window size selection. This strategy removed the noise without loosing valuable information. In some cases, it was found preferable to split the chromatogram in different elution regions, and apply the Durbin-Watson test and the Savitsky-Golay smoother to each region, separately. The derivatives allowed obtaining estimates of both peak parameters and the corresponding ranges for each eluting compound to be used in the deconvolution. An algorithm oriented to compare peaks from different chromatograms is also presented to perform deconvolution, using information from several related chromatograms.

Algorithms↗

An alternative viewpoint of the similarities and differences of SVD and FT deconvolution algorithms used for quantitative MR perfusion studies.

Quantitative cerebral blood flow (CBF) values can be determined from residue function estimates obtained from magnetic resonance dynamic susceptibility contrast (DSC) perfusion studies using a variety of deconvolution approaches. However, there are significant differences between the CBF estimates obtained, differences that are not simply due to minor details of the implementation of the algorithms. The standard singular value decomposition (sSVD) shows a variation of CBF values with arterial-tissue delay (ATD) not present with the Fourier transform deconvolution algorithm. Fourier transform deconvolution and the newly suggested delay-invariant SVD algorithm implementations provide CBF estimates whose accuracy changes with tissue mean transit times (MTTs). Techniques combining sSVD with deliberate ATD manipulation have been proposed to compensate for this inaccuracy. Other studies indicate that CBF changes related to slice position in a multislice study, and other experimental factors, can be reduced using interpolative deconvolution algorithms. In this review, we use both time-domain and frequency-domain analysis to show the underlying theoretical relationships between these many approaches to obtain "the best" CBF estimate. This model allows us to better understand the similarities and differences, advantages and disadvantages between these variants of the deconvolution algorithms used in DSC perfusion studies.

Algorithms↗

Image deconvolution in spherical aberration-corrected high-resolution transmission electron microscopy.

The method of image deconvolution developed previously for FEG high-resolution transmission electron microscope (HRTEM) without a spherical aberration (C(s)) corrector was for the first time applied to FEG HRTEM with a C(s)-corrector. The principle and the procedure of image deconvolution are briefly described. Four qualified [1 1 0] images of Si were selected from a through-focus series to perform image deconvolution. The projected potential is successfully derived from all the images, and the obtained "dumbbell" structure maps of Si [1 1 0] are in good agreement with the calculated potential map. The criterion of selecting qualified images for performing image deconvolution is indicated. The possibility of applying image deconvolution to defect study and to ab initio crystal structure determination is discussed.

Journal Article↗

Semi-sparse deconvolution robust to uncertainties in the impulse responses.

The received signal in ultrasonic pulse-echo inspection can be modeled as a convolution between an impulse response, or prototype, and the reflection sequence that is the impulse characteristic of the inspected object. Deconvolution aims at approximately inverting this process to improve the time resolution so that the overlap between echoes from closely spaced reflectors becomes small. For the relatively high contrast reflection sequences often found in non-destructive testing applications, semi-sparse deconvolution algorithms will typically yield better results than the classical Wiener filter solution. However, this requires a prototype that is a good representative for all echo responses found in the signals. Since, in practice, the prototype is often chosen by the operator directly from the inspection data, the prototype may incidentally be a bad representative for modeling the echoes for the remaining part of the object. Because of the sensitivity to deviations in the prototype this can yield deconvolution results with poor reproducibility. This paper presents a new semi-sparse deconvolution algorithm that is robust to deviations in the prototype. The new robust algorithm is based on a modification of an earlier presented non-robust semi-sparse algorithm. The robustness is obtained by including a stochastic model of the variations in the prototypes to the signal model when deriving the algorithm. Experiments performed using simulated data verify that the robust algorithm is less sensitive to deviations in the prototypes compared to the non-robust version of the algorithm and show that the proposed algorithm yields better estimates than its non-robust version and the Wiener filter in scenarios for which the algorithm was derived. Results using real ultrasonic data further show that the algorithm can be useful in practical scenarios where similar deconvolution results are required from slightly different echoes.

Journal Article↗

Estimating DOC regime in a wastewater treatment plant by UV deconvolution.

A UV-deconvolution method was modified, and applied to estimation of dissolved organic carbon (DOC) along a municipal wastewater treatment plant (WWTP) in Catalonia, Spain. One grab sample was taken every 2h at four sampling points, for 1 week (336 samples), in order to characterise day/night and weekday/weekend DOC regimes along the plant. Samples were centrifuged before DOC determination or estimation. Four components were selected for describing wastewater composition and spectra. Reference spectra for these components were taken from the literature and proved to correctly explain the sample spectra. A two-step deconvolution method was developed, which avoided negative nitrate coefficients while keeping deconvolution error low. The calibration file for DOC estimation was determined by analysing DOC and acquiring UV spectra from 48 samples. DOC values were correlated to UV spectra by multiple linear regression. Determination coefficient and standard error were comparable to the values found in the literature. In raw or diluted samples with an absorbance between 2.0 and 2.5, DOC was probably underestimated by the method. This points to some nonlinearity for absorbances above 2.0, rather than the 2.5 limit suggested by the original method. DOC calculation through UV deconvolution allowed for the estimation of DOC regime along the WWTP. Time bands for higher and lower DOC concentrations were determined and characterised at each sampling point, for weekdays and the weekend. Except for the plant effluent, clear time bands were found. In effluent, DOC was always low, and very small oscillations were detected, due to DOC removal and intense mixing in the biological process. DOC profiles at each point are discussed in this paper. The modified deconvolution method has proved to be an accurate and efficient technique for estimating DOC of a large number of raw and diluted samples.

Calibration↗

Widefield deconvolution epifluorescence microscopy combined with fluorescence in situ hybridization reveals the spatial arrangement of bacteria in sponge tissue.

Widefield deconvolution epifluorescence microscopy (WDEM) combined with fluorescence in situ hybridization (FISH) was performed to identify and characterize single bacterial cells within sections of the mediterranean sponge Chondrosia reniformis. Sponges were embedded in paraffin wax or plastic prior to the preparation of thin sections, in situ hybridization and microscopy. Serial digital images generated by widefield epifluorescence microscopy were visualized using an exhaustive photon reassignment deconvolution algorithm and three-dimensional rendering software. Computer processing of series of images taken at different focal planes with the deconvolution technique provided deblurred three-dimensional images with high optical resolution on a submicron scale. Results from the deconvolution enhanced widefield microscopy were compared with conventional epifluorescent microscopical images. By the application of the deconvolution algorithm on digital image data obtained with widefield epifluorescence microscopy after FISH, the occurrence and spatial arrangement of Desulfovibrionaceae closely associated with micropores of Chondrosia reniformis could be visualized.

Algorithms↗

Deconvolution of combinatorial libraries for Drug discovery: theoretical comparison of pooling strategies.

Synthesis and testing of mixtures of compounds in a combinatorial library allow much greater throughput than synthesis and testing of individual compounds. When mixtures of compounds are screened, however, the possibility exists that the most active compound will not be identified. The specific strategies employed for pooling and deconvolution will affect the likelihood of success. We have used a nucleic acid hybridization example to develop a theoretical model of library deconvolution for a library of more than 250,000 compounds. This model was used to compare various strategies for pooling and deconvolution. Simulations were performed in the absence and presence of experimental error. We found iterative deconvolution to be most reliable when active molecules were assigned to the same subset in early rounds. Reliability was reduced only slightly when active molecules were assigned randomly to all subsets. Iterative deconvolution with as many as 65,536 compounds per subset did not drastically reduce the reliability compared to one-at-a-time testing. Pooling strategies compared using this theoretical model are compared experimentally in an accompanying paper.

Base Sequence↗

Discovery of novel pyridinopolyamines with potent antimicrobial activity: deconvolution of mixtures synthesized by solution-phase combinatorial chemistry.

A 1638-member pyridinopolyamine library, consisting of 13 sublibraries of 126 members prepared by a solution-phase approach, was completely deconvoluted from orthogonally protected intermediates by a combination of iterative and positional scanning procedures. Antibacterial assays against Streptococcus pyogenes and Escherichia coli imp- and a Candida albicans yeast specificity assay were employed to follow the activity of sublibraries. Screening of the 13 sublibraries, which were prepared by a synthetic method that places the differentiating functionality in a selected position A (secondary amine), at the end of the synthesis (fix last), provided several first-round activities. Subsequently, six single pyridinopolyamines (2-7) were prepared where the first-round winner, a hydrogen atom, is in the first deconvoluted position and the remaining three positions contained the same functionalities. The range of antibacterial and yeast activities of these single compounds suggested that a more active and selective compound may be discovered by completely deconvoluting the first-round active sublibraries. Pyridinopolyamine positions B (secondary benzylamine) and C (primary benzylamine) were then sequentially positionally scanned with a set of six meta-substituted benzyl functionalities to generate two sets of second/third-round sublibraries, containing 21 or 36 compounds in each sublibrary, respectively. High-throughput screening yielded sublibraries 15, 18, and 21 with MICs of 1-5 microM against S. pyogenes and E. coli imp-. Using rounds 1 and 2/3 screening data, two sets of single compounds (22-27) and (28-32) with the combination of m-(trifluoromethyl)-benzyl group at position C and m-(trifluoromethyl)benzyl or m-methylbenzyl group at position B with position D (primary benzylamine) fixed were synthesized in the fourth round deconvolution. Subsequently, broader screening of deconvoluted compounds against a tier II panel of wild-type bacteria identified eight compounds (5, 7, 27, and 29-32) with approximately 100-fold greater selectivity for Gram-positive than Gram-negative bacteria. Thus, S. pyogenes, S. pyogenes (wild-type), Streptomyces aureus, and Enterococcus faecalis were inhibited at MICs of 1-12 microM, whereas MICs for E. coli, Klebsiella pneumoniae, Proteus vulgaris, and Pseudomonas aeruginosa were > 100 microM. These eight compounds were not active (> 100 microM) against fungus C. albicans.

Anti-Bacterial Agents↗

Blind deconvolution of 3D transmitted light brightfield micrographs.

The blind deconvolution algorithm for 3D transmitted light brightfield (TLB) microscopy, published previously [Holmes et al. Handbook of Biological Confocal Microscopy (1995)], is summarized with example images. The main emphasis of this paper is to discuss more thoroughly the importance and usefulness of this method and to provide more detailed evidence, some being quantitative, of its necessity. Samples of horseradish peroxidase (HRP)-stained pyramidal neurones were prepared and evaluated for the ability to see fine structures clearly, including the dendrites and spines. It is demonstrated that the appearance of fine spine structure, and means of identifying spine categories, is made possible by using blind deconvolution. A comparison of images of the same sample from reflected light confocal microscopy, which is the conventional light microscopic way of viewing the 3D structure of these HRP-stained samples, shows that the blind deconvolution method is far superior for clearly showing the structure with less distortion and better resolution of the spines. The main significance of this research is that it is now possible to obtain clear images of 3D structure by light microscopy of absorbing stains. This is important because the TLB microscope is probably the most widely used modality in the life-science laboratory, yet, until now, there has been no reliable means for it to provide visualization of 3D structure clearly. The main importance of the blind deconvolution approach is that it obviates the need to measure the point spread function of the optical system, so that it now becomes realistic to provide a 3D light microscopic deconvolution method that can be pervasively used by microscopists.

Algorithms↗