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PubMed2004

A novel inhibitor of vacuolar ATPase, FR167356, which can discriminate between osteoclast vacuolar ATPase and lysosomal vacuolar ATPase.

1 Vacuolar ATPase (V-ATPase) has been proposed as a drug target in lytic bone diseases. Studies of bafilomycin derivatives suggest that the key issue regarding the therapeutic usefulness of V-ATPase inhibitors is selective inhibition of osteoclast V-ATPase. Previous efforts to develop therapeutic inhibitors of osteoclast V-ATPase have been frustrated by a lack of synthetically tractable and biologically selective leads. Therefore, we tried to find novel potent and specific V-ATPase inhibitors, which have new structural features and inhibition selectivity, from random screening using osteoclast microsomes. Finally, a novel V-ATPase inhibitor, FR167356, was obtained through chemical modification of a parental hit compound. 2 FR167356 inhibited not only H+ transport activity of osteoclast V-ATPase but also H+ extrusion from cytoplasm of osteoclasts, which depends on the V-ATPase activity. As expected, FR167356 remarkably inhibited bone resorption in vitro. 3 FR167356 also showed inhibitory effects on other V-ATPases, renal brush border V-ATPase, macrophage microsome V-ATPase and lysosomal V-ATPase. However, FR167356 was approximately seven-fold less potent in inhibiting lysosomal V-ATPase compared to osteoclast V-ATPase. Moreover, LDL metabolism in cells, which depends on acidification of lysosome, was blocked merely at higher concentration than bone resorption, suggesting that FR167356 inhibits V-ATPase of osteoclast ruffled border membrane still more selectively than lysosome at the cellular level. 4 These results from the experiments seem to indicate that osteoclast V-ATPase may be different from lysosomal V-ATPase in respect of their structure. 5 FR167356 had a novel chemical structural feature as well as inhibitory characteristics distinctly different from any previously known V-ATPase inhibitor family. Therefore, FR167356 is thought to be a useful tool for estimating the essential characteristics of V-ATPase inhibitors for drug development.

Animals↗
PubMed2005

Multimodality computerized diagnosis of breast lesions using mammography and sonography.

RATIONALE AND OBJECTIVES: The purpose of this study is to investigate the use of computer-extracted features of lesions imaged by means of two modalities, mammography and breast ultrasound, in the computerized classification of breast lesions. MATERIAL AND METHODS: We performed computerized analysis on a database of 97 patients with a total of 100 lesions (40 malignant, 40 benign solid, and 20 cystic lesions). Mammograms and ultrasound images were available for these breast lesions. There was an average of three mammographic images and two ultrasound images per lesion. Based on seed points indicated by a radiologist, the computer automatically segmented lesions from the parenchymal background and automatically extracted a set of characteristic features for each lesion. For each feature, its value averaged over all images pertaining to a given lesion was input to a Bayesian neural network for classification. We also investigated different approaches to combine image-based features into this by-lesion analysis. In that analysis, mean, maximum, and minimum feature values were considered for all images representing a lesion. We considered performance by using a leave-one-lesion-out approach, based on image features from mammography alone (two to five features), ultrasound alone (three to four features), and a combination of features from both modalities (three to five features total). RESULTS: For the classification task of distinguishing cancer from other abnormalities in a lesion-based analysis by using a single modality, areas under the receiver operating characteristic curves (A(z) values) increased significantly when the computer selected the manner (mean, minimum, or maximum) in which image-based features were combined into lesion-based features. The highest performance was found for lesion-based analysis and automated feature selection from mean, maximum, and minimum values of features from both modalities (resulting in a total of four features being used). That A(z) value for the task of distinguishing cancer was 0.92, showing a statistically significant increase over that achieved with features from either mammography or ultrasound alone. CONCLUSION: Computerized classification of cancer significantly improved when lesion features from both modalities were combined. Classification performance depended on specific methods for combining features from multiple images per lesion. These results are encouraging and warrant further exploration of computerized methods for multimodality imaging.

Breast Neoplasms↗
PubMed2003

Columns for complex visual object features in the inferotemporal cortex: clustering of cells with similar but slightly different stimulus selectivities.

Cells in the inferotemporal cortex (area TE) selectively respond to complex visual object features and those that respond to similar features cluster in a columnar region elongated vertical to the cortical surface. What are the functional roles of the column structure in the inferotemporal cortex? Selectivity of cells within a column is similar but not identical. If we emphasize the similarity among cells within a column, we can regard the columns as units for description of object features. The variety of stimulus selectivity in a column may work as a tool to disregard subtle changes in input images when the system is directed to invariant recognition. Alternatively, if we emphasize the differences in selectivity of cells within a column, the columns can be compared to differential amplifiers, each of which represents variety within a group of features. The enormous number of objects present in nature can be efficiently described by combining outputs of the multiple differential amplifiers in the inferotemporal cortex. The two modes may work in parallel, with a graded balance changing according to the behavioral context. Determining whether or not these hypotheses are valid will require further studies.

Animals↗
PubMed2004

The integration of multiple stimulus features by V1 neurons.

We investigated how V1 neurons integrate two stimulus features by presenting stimuli from a stimulus set made up of all combinations of eight different directions of motion and nine binocular disparities. We investigated the occurrence and shape of the resulting joint tuning function. Among V1 neurons, approximately 80% were jointly tuned for disparity with orientation or direction. The joint tuning function of all jointly tuned neurons was separable into distinct tuning for disparity on the one hand, and orientation or direction tuning on the other. The degree of separability and the mutual information between the stimulus and the firing rates were strongly correlated. The mutual information of jointly tuned neurons when both features were decoded together was highly correlated with the mutual information when the two features were decoded separately, and the information was then summed. Jointly tuned neurons were just as good at representing information about single features as neurons tuned for only a single feature. The tuning properties of most jointly tuned neurons did not dynamically evolve over time, nor did jointly tuned neurons respond earlier than neurons tuned for only a single feature. The response selectivity of V1 neurons is low and decreases the information that a neuron represents about a stimulus. Together these results suggest that distinct stimulus features are integrated very early in visual processing. Furthermore, V1 generates a distributed representation through low response selectivity that avoids the curse of dimensionality by using separable joint tuning functions.

Animals↗
PubMed2000

Experimental verification of a predicted, hitherto unseen separation selectivity pattern in the nonaqueous capillary electrophoretic separation of weak base enantiomers by octakis (2,3-diacetyl-6-sulfato)-gamma-cyclodextrin.

The capillary electrophoretic separation of cationic enantiomers with single-isomer multivalent anionic resolving agents was reexamined with the help of the charged resolving agent migration model. Three general model parameters were identified that influence the shape of the separation selectivity and enantiomer mobility difference curves: parameter b, the binding selectivity (K(RCD)/K(SCD)), parameter s, the size selectivity (mu0(RCD)/mu0(SCD)), and parameter a, the complexation-induced alteration of the analyte's mobility (mu0(RCD)/mu0). In addition to the previously observed discontinuity in separation selectivity that occurs as mu(eff) of the less mobile enantiomer changes from cationic to anionic, a new feature, a separation selectivity maximum was predicted to occur in the resolving agent concentration range where both enantiomers migrate cationically provided that (i) K(RCD)/K(SCD) <1 and mu0(RCD)/mu0(SCD) >1 and (K(RCD)mu0(RCD))/(K(SCD)mu0(SCD)) > 1, or (ii) K(RCD)/K(SCD) >1 and mu0(RCD)/mu0(SCD) <1 and (K(RCD)mu0(RCD))/(K(SCD)mu0(SCD)) <1. This hitherto unseen separation selectivity pattern was experimentally verified during the nonaqueous capillary electrophoretic separation of the enantiomers of four weak base analytes in acidic methanol background electrolytes with octakis(2,3-diacetyl-6-sulfato)-gamma-cyclodextrin (ODAS-gammaCD) as resolving agent.

Albuterol↗
PubMed1974

Electrochemical properties of hydrated cation-selective glass membrane. A model of K+ and Na+ transport.

Electrochemical properties of cation-selective glass microelectrodes made from NAS(27-04) were studied. There was a marked fall in electrical resistance of the microelectrodes stored in 3 M KCl solution (aging). The resistance was in the range of 2 x 10(7) to 10(9) Omega, which were much lower than those estimated from the electrical resistivity of dry glass for the equivalent dimensions of microelectrode working tips. This fall in resistance was accompanied by an increase in microelectrode selectivity for K(+). The low resistance and increased K(+) selectivity are desirable features that make the microelectrode more suitable for application to biologic studies. The changes in microelectrode resistance and selectivity were interpreted to be due to hydration of the entire thickness of the glass membrane, resulting in a change in the field strength of anionic sites and formation of ionic channels in the glass membrane. Thus, the fall in resistance is explained by decrease in energy barrier, which is equivalent to the activation energy of interaction between the cations and anionic sites in the glass membrane. Some of the microelectrodes showed a transient depolarization that resembled the action potential of a biological membrane. This transient depolarization was associated with the changes in microelectrode resistance and selectivity. The transient depolarizations suggest the temporary development of wide channels in the membrane permitting free movement of hydrated cations according to the bulk electrochemical gradient.

Action Potentials↗
PubMed2006

Selection of allosteric beta-lactamase mutants featuring an activity regulation by transition metal ions.

Libraries of phage-displayed beta-lactamase mutants in which up to three loops have been engineered by genetic introduction of random peptide sequences or by randomization of the wild-type sequence have been submitted to selection protocols designed to find mutants in which binding of transition metal ions to the engineered secondary binding site leads to significant effects on the enzymatic activity. A double-selection protocol was applied: The phage-displayed libraries were first selected for transition metal ions affinity by panning on IMAC support, then a second selection step was applied to isolate mutants that have retained significant catalytic activity. The analysis of the kinetic properties of mutants in the presence of nickel, copper, or zinc ions allowed isolation of a few mutants whose activity was either enhanced or inhibited by factors up to three and >10, respectively, in a metal-specific manner. A remarkable mutant exhibiting differential allosteric regulation depending on the metal was found. Its activity was activated by nickel ion binding, inhibited by cupric ion binding, and nearly unaffected by zinc ions. These observations point to an interesting potential for up- or down-regulation of activity within a monomeric enzyme by binding to an "allosteric site" relatively remote from the active site.

Allosteric Regulation↗
PubMed1999

Effects of box features on spine loading during warehouse order selecting.

Low back disorders in distribution centres or warehouses have been identified as an area of elevated risk in many industries. The task of an order selector requires workers manually to lift boxes from storage bins to a mobile pallet. This study explored the effect of box features and box location when lifting from a pallet in a storage bin upon spine loading. Ten experienced warehouse workers were asked to lift boxes from a pallet while the size, weight, handle features and location of the box on a pallet were changed. An EMG-assisted model was employed to assess spine compression, lateral shear and anterior-posterior shear during the lifts. The position from which the worker lifted a box on a pallet had the most profound effect on spine loading while the lower level of the pallet represented the greatest loadings on the spine. Box weight did not appear to be a feasible means of controlling spine loading unless its position on the pallet could also be controlled. The inclusion of handles had an effect similar to reducing the box weight by 4.5 kg, whereas box size did not effectively affect spine loading. The mechanisms by which these factors affect spine loading are discussed.

Adult↗
PubMed2004

Visual categorization and the inferior temporal cortex.

We investigated the effects of categorization on the representation of stimulus features in combined psychophysical-electrophysiological experiments. We used parameterized line drawings of faces and fish as stimuli, and we varied the relevance of the different features for the categorization task. The psychophysical and electrophysiological data support an exemplar-based framework for visual object recognition. We recorded from visual neurons in the anterior inferior temporal (IT) cortex of macaque monkeys, while they were performing a categorization task. The visual neurons did not respond selectively to one stimulus set, or to one category. The majority of the anterior IT feature selective neurons were tuned for features that were diagnostic for the categorization task. We argue that this fine-tuning of the neurons reflects the perceptual sensitization to the diagnostic features.

Analysis of Variance↗
PubMed1996

Efficient Biologically-based Pattern-recognizing Networks.

A biologiclly-motivated classifying neural network which is based on the feature extraction scheme found in the visual cortex is suggested. A special process is proposed for grading and automatically selecting the "best" features for specific recognition tasks. Ranking is based on a feature's calculated discriminating ability, such that a given class is separated from each and every other class by a given amount. The outcome is a net with less computational complexity than other neural nets, yet one which is more biologically plausible.The main motivation for constructing a reduced net is that the complex circuitry of the brain deals with a huge number of patterns, while a machine-based recognition system usually deals with a limited number of patterns. Results show that feature reduction is drastic and that very compact nets, of the order of tens of neurons, can be used to classify patterns, even in a noisy environment. Copyright 1996 Elsevier Science Ltd

Journal Article↗
PubMed2005

An in vitro assay for the selective endoplasmic reticulum associated degradation of an unglycosylated secreted protein.

The endoplasmic reticulum (ER) represents the first compartment into which nascent secreted proteins traffic, and not coincidentally the ER lumen houses a high concentration of factors that facilitate protein folding, such as molecular chaperones. To off-set the potentially lethal consequences of mis-folded secreted protein accumulation, aberrant proteins may be selected for degradation via a process known as ER associated degradation (ERAD). After their selection ERAD substrates are retro-translocated back to the cytoplasm and then degraded by the 26S proteasome. Key features of the selection, retro-translocation, and degradation steps that constitute the ERAD pathway were elucidated through the development of an in vitro ERAD assay. In this assay the fates of two yeast proteins can be distinguished after their translocation, or import into ER-derived microsomes. Whereas a wild type, glycosylated protein ("Gp(alpha)F") is stable, a non-glycosylated version of the same protein ("p(alpha)F") is rapidly degraded when microsomes containing radiolabeled forms of these substrates are incubated in cytosol and ATP. The purpose of this chapter is first to discuss the experimental findings from the use of the in vitro assay, and then to describe the assay in detail. Finally, future potential uses of the in vitro system are illustrated.

Endoplasmic Reticulum↗
PubMed1999

Discrimination of MR images of breast masses with fractal-interpolation function models.

RATIONALE AND OBJECTIVES: The authors evaluated the feasibility of using statistical fractal-dimension features to improve discrimination between benign and malignant breast masses at magnetic resonance (MR) imaging. MATERIALS AND METHODS: The study evaluated MR images of 32 malignant and 20 benign breast masses from archived data at the University of Pennsylvania Medical Center. The test set included four cases that were difficult to evaluate on the basis of border characteristics. All diagnoses had been confirmed at excisional biopsy. The fractal-dimension feature was computed as the mean of a sample space of fractal-dimension estimates derived from fractal interpolation function models. To evaluate the performance of the fractal-dimension feature, the classification effectiveness of five expert-observer architectural features was compared with that of the fractal dimension combined with four expert-observer features. Feature sets were evaluated with receiver operating characteristic analysis. Discrimination analysis used artificial neural networks and logistic regression. Robustness of the fractal-dimension feature was evaluated by determining changes in discrimination when the algorithm parameters were perturbed. RESULTS: The combination of fractal-dimension and expert-observer features provided a statistically significant improvement in discrimination over that achieved with expert-observer features alone. Perturbing selected parameters in the fractal-dimension algorithm had little effect on discrimination. CONCLUSION: A statistical fractal-dimension feature appears to be useful in distinguishing MR images of benign and malignant breast masses in cases where expert radiologists may have difficulty. The statistical approach to estimating the fractal dimension appears to be more robust than other fractal measurements on data-limited medical images.

Algorithms↗
PubMed1993

Proteinuria selectivity index--prognostic value in lipoid nephrosis and related diseases.

In order to predict the steroid response in lipoid nephrosis (LN), we studied age, sex, proteinuria level, histological features and proteinuria selectivity index (SI; ratio between IgG and transferrin clearances) in 52 LN cases (minimal-change disease: n = 39; focal glomerulosclerosis+IgM nephropathy: n = 13). The multivariate analysis showed that age, sex and proteinuria level were not contributive, whereas histology and SI were. The predictive value of SI was much higher than that of histological type (McFadden's r2: 47% vs. 22%, p < 0.001). Thus, SI should be systemically assessed in idiopathic nephrotic syndrome for reviewing the pathologic classification obtained by histology. However, if its prognostic value is lower than that of selectivity, initial renal biopsy remains necessary for diagnosis in adults.

Adolescent↗
PubMed2003

Should there be a size limit for elective nephron-sparing surgery?

Nephron-sparing surgery (NSS) is a mandatory procedure for patients with solid renal masses who also have coexisting urologic or medical conditions that pose a threat to overall renal function. The excellent results observed with this procedure have led patients with normal contralateral kidneys to choose elective NSS as a treatment modality. However, the optimal selection criteria for NSS have not yet been defined. We review the developments in and recent results of NSS and discuss features critical in selecting patients for this procedure on an elective basis. Current data clearly support the use of elective NSS for localized solid renal masses under 4 cm in size.

Carcinoma, Renal Cell↗
PubMed2000

Accident reduction factors and causal inference in traffic safety studies: a review.

Accident reduction factors are used to predict the change in accident occurrence which a countermeasure can be expected to cause. Since ethical and legal obstacles preclude the use of randomized experiments when evaluating traffic safety improvements, empirical support for the causal effectiveness of accident countermeasures comes entirely from observational studies. Drawing on developments in causal inference initiated by Donald Rubin, it is argued here that the mechanism by which sites are selected for application of a countermeasure should be included as part of a study's data model, and that when important features of the selection mechanism are neglected, existing methods for estimating accident reduction factors become inconsistent. A promising, but neglected, way out of these difficulties lies in developing rational countermeasure selection methods which also support valid causal inference of countermeasure effects.

Accidents, Traffic↗
PubMedDate not supplied

Thyroid nodules with microfollicular findings reported on fine-needle aspiration: invariably surgical treatment?

OBJECTIVE: To assess whether fine-needle aspiration (FNA) can be a diagnostic procedure for identification of malignant thyroid nodules when the smear reveals the cytologic (microfollicular) finding of "follicular neoplasm" (FN). METHODS: We reviewed a group of 79 patients with FN who underwent surgical treatment and evaluated the possible significance of the cytologic features of anisokaryosis, amount of colloid, and nuclear overlapping (NO). We excluded from the study those patients with FN who had other highly suspicious cytologic features, such as sporadic grooves and sporadic pseudonucleoli. RESULTS: In our series of 79 patients with FN at FNA, we found anisokaryosis in 49.4%, scant or no colloid in 31.6%, and NO in 5.1%. Thyroid cancer (TC)--follicular cancer or follicular variant of papillary cancer--was found in six patients (7.6%): four patients (10.2%) with anisokaryosis and two patients (5.0%) without anisokaryosis (P<0.5; chi2 test). All the patients with malignant lesions had scant or no colloid in the smears, and four of them were those with NO. Therefore, TC was found in 24.0% of nodules with scant or no colloid versus 0.0% of nodules with abundant colloid (P<0.001; chi2 test), and TC was found in 100% of nodules with NO versus 2.6% of nodules without NO (P<0.001; chi2 test). CONCLUSION: Because of the high prevalence of thyroid nodules and the frequency of FN at FNA, the number of unnecessary interventions may be very high. FNA cannot be diagnostic in FN because invasion of the tumor capsule and blood vessels can be evidenced only at histologic examination. Although it is common opinion that microfollicular nodules should be surgically treated, simple cytologic features can help select patients at low risk who can be monitored and reassessed clinically and by FNA. Patients with high-risk cytologic features such as NO should be advised that a total thyroidectomy may be necessary.

Adenocarcinoma, Follicular↗
PubMedDate not supplied

Authoring systems. Finding the right tool for your courseware development project.

Many factors must be considered when selecting an appropriate tool for courseware development. An overview of development tools is provided. Based on the authors' experience in selecting an authoring system for a recent courseware development project, a practical process for selecting the appropriate tool is described. Considerations when determining the general needs for an authoring system and evaluating specific authoring systems' features are outlined. Selection considerations are also discussed.

Curriculum↗
PubMed1998

Consciousness and the structure of neuronal representations.

The hypothesis is defended that brains expressing phenomenal awareness are capable of generating metarepresentations of their cognitive processes, these metarepresentations resulting from an iteration of self-similar cortical operations. Search for the neuronal substrate of awareness therefore converges with the search for the nature of neuronal representations. It is proposed that evolved brains use two complementary representational strategies. One consists of the generation of neurons responding selectively to a particular constellation of features and is based on selective recombination of inputs in hierarchically structured feedforward architectures. The other relies on the dynamic association of feature-specific cells into functionally coherent cell assemblies that, as a whole, represent the constellation of features defining a particular perceptual object. Arguments are presented that favour the notion that the metarepresentations supporting awareness are established in accordance with the second strategy. Experimental data are reviewed that are compatible with the hypothesis that evolved brains use assembly codes for the representation of contents and that these assemblies become organized through transient synchronization of the discharges of associated neurons. It is argued that central states favouring the formation of assembly-based representations are similar to those favouring awareness.

Animals↗