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At least 217 records · Page 12Linked to original sources

Application of predictive QSAR models to database mining: identification and experimental validation of novel anticonvulsant compounds.

We have developed a drug discovery strategy that employs variable selection quantitative structure-activity relationship (QSAR) models for chemical database mining. The approach starts with the development of rigorously validated QSAR models obtained with the variable selection k nearest neighbor (kNN) method (or, in principle, with any other robust model-building technique). Model validation is based on several statistical criteria, including the randomization of the target property (Y-randomization), independent assessment of the training set model's predictive power using external test sets, and the establishment of the model's applicability domain. All successful models are employed in database mining concurrently; in each case, only variables selected as a result of model building (termed descriptor pharmacophore) are used in chemical similarity searches comparing active compounds of the training set (queries) with those in chemical databases. Specific biological activity (characteristic of the training set compounds) of external database entries found to be within a predefined similarity threshold of the training set molecules is predicted on the basis of the validated QSAR models using the applicability domain criteria. Compounds judged to have high predicted activities by all or the majority of all models are considered as consensus hits. We report on the application of this computational strategy for the first time for the discovery of anticonvulsant agents in the Maybridge and National Cancer Institute (NCI) databases containing ca. 250,000 compounds combined. Forty-eight anticonvulsant agents of the functionalized amino acid (FAA) series were used to build kNN variable selection QSAR models. The 10 best models were applied to mining chemical databases, and 22 compounds were selected as consensus hits. Nine compounds were synthesized and tested at the NIH Epilepsy Branch, Rockville, MD using the same biological test that was employed to assess the anticonvulsant activity of the training set compounds; of these nine, four were exact database hits and five were derived from the hits by minor chemical modifications. Seven of these nine compounds were confirmed to be active, indicating an exceptionally high hit rate. The approach described in this report can be used as a general rational drug discovery tool.

Amides↗

Three dimensional atomic model and experimental validation for the ATP-Regulated Module (ARM) of the atrial natriuretic factor receptor guanylate cyclase.

Atrial natriuretic factor (ANF) receptor guanylate cyclase (ANF-RGC) is a single chain transmembrane-spanning protein, containing both ANF binding and catalytic activities. ANF binding to the extracellular receptor domain activates the cytosolic catalytic domain, generating the second messenger cyclic GMP. Obligatory in this activation process is an intervening transduction step, which is regulated by the binding of ATP to the cyclase. The partial structural motif of the ATP binding domain of the cyclase has been elucidated and has been termed ATP Regulatory Module (ARM). The crystal structures of the tyrosine kinase domains of the human insulin receptor and haematopoietic cell kinase were used to derive a homology-based model of the ARM domain of ANF-RGC. The model identifies the precise configuration of the ATP-binding pocket in the ARM domain, accurately represents its ATP-dependent features, and shows that the ATP-dependent transduction phenomenon is a two-step mechanism. In the first step, ATP binds to its pocket and changes its configuration; in the second step, via an unknown protein kinase, it phosphorylates the cyclase for its full activation.

Adenosine Triphosphate↗

Three dimensional atomic model and experimental validation for the ATP-Regulated Module (ARM) of the atrial natriuretic factor receptor guanylate cyclase.

Atrial natriuretic factor (ANF) receptor guanylate cyclase (ANF-RGC) is a single chain transmembrane-spanning protein, containing both ANF binding and catalytic activities. ANF binding to the extracellular receptor domain activates the cytosolic catalytic domain, generating the second messenger cyclic GMP. Obligatory in this activation process is an intervening transduction step, which is regulated by the binding of ATP to the cyclase. The partial structural motif of the ATP binding domain of the cyclase has been elucidated and has been termed ATP Regulatory Module (ARM). The crystal structures of the tyrosine kinase domains of the human insulin receptor and haematopoietic cell kinase were used to derive a homology-based model of the ARM domain of ANF-RGC. The model identifies the precise configuration of the ATP-binding pocket in the ARM domain, accurately represents its ATP-dependent features, and shows that the ATP-dependent transduction phenomenon is a two-step mechanism. In the first step, ATP binds to its pocket and changes its configuration; in the second step, via an unknown protein kinase, it phosphorylates the cyclase for its full activation.

Adenosine Triphosphate↗

Experimental validation of videoendoscopic isolation and occlusion of the bloodflow in open ductus arteriosus.

The potentialities of videoendoscopic isolation and extravasal occlusion of large vessels with titanium clips were studied in acute and chronic experiments on 36 animals and anatomic studies on 5 human cadavers. The number and optimal sites for insertion of trocars and disposition of instruments and thoracoscope for isolation of open ductus arteriosus were determined. A method for videoendoscopic isolation of vessels is developed and possible complications and measures for their liquidation evaluated. High efficiency and low traumatism of videoendoscopic operation are proven under condition of strict adherence to methodology and sufficient qualification of the surgeon in endosurgical manipulations.

Animals↗

General cognitive slowing in the nonlexical domain: an experimental validation.

Older and younger adults were tested on 4 nonlexical tasks: choice reaction time, letter classification, mental rotation, and abstract matching. A positively accelerated relation was observed between older and younger adults' latencies. Consistent with general slowing, the relation observed with the same subjects in each condition was more than 3 times as precise as in a comparable meta-analysis. Further analyses compared the ability of various models to describe the present data and also to predict the data on the basis of parameters estimated from a previous meta-analysis. Compared with linear models, the information-loss and overhead models provided more accurate accounts of general cognitive slowing in the nonlexical domain.

Adult↗

Experimental validation of a predicted feedback loop in the multi-oscillator clock of Arabidopsis thaliana.

Our computational model of the circadian clock comprised the feedback loop between LATE ELONGATED HYPOCOTYL (LHY), CIRCADIAN CLOCK ASSOCIATED 1 (CCA1) and TIMING OF CAB EXPRESSION 1 (TOC1), and a predicted, interlocking feedback loop involving TOC1 and a hypothetical component Y. Experiments based on model predictions suggested GIGANTEA (GI) as a candidate for Y. We now extend the model to include a recently demonstrated feedback loop between the TOC1 homologues PSEUDO-RESPONSE REGULATOR 7 (PRR7), PRR9 and LHY and CCA1. This three-loop network explains the rhythmic phenotype of toc1 mutant alleles. Model predictions fit closely to new data on the gi;lhy;cca1 mutant, which confirm that GI is a major contributor to Y function. Analysis of the three-loop network suggests that the plant clock consists of morning and evening oscillators, coupled intracellularly, which may be analogous to coupled, morning and evening clock cells in Drosophila and the mouse.

Arabidopsis↗

[Experimental validation of DXA and MRI-based bone density measurement by ash-method].

Different methods have been established for bone density measurements such as dual energy X-ray absorptiometry (DXA), quantitative computertomography (QCT), and scintigraphy (VQ-Scan). There are hints that magnetic resonance imaging (MRI) might become a new option for the evaluation of bone density. The aim of this study was to investigate correlations between MRI vs. DXA and MRI vs. mineral content of lumbar vertebrae. Data were obtained from ten lumbar vertebral bodies of cattles. The T-1 MRI-sequences SE, PS, and the T-2 Sequence STIR were used for analysis. Total pixel numbers and a pixel per area ratio were determined. Values were compared to DXA-measurements, to the wet weight, and to separated measurements of the spongious, trabecular, and total mineral content of the vertebral body after ashing. We found correlations between DXA (g/vertebral body) vs. mineral content by ash-method (0.918; p < 0.01), DXA vs. MRI (SE-sequence) (-0.872; p < 0.01), and MRI (SE-sequence) vs. mineral content (0.775; p < 0.01). No correlations were found between PS- or STIR-sequences and the ash-method. This study shows that the determination of the bone mineral content of vertebrae is possible applying MRI in the T1-weighted SE-sequence. Without radiation, the MRI provides additionally early detection of trabecular lesions, fractures, and deformities at the spine, without other diagnostic procedures becoming necessary.

Absorptiometry, Photon↗

[Cardiac MR flowmetry: experimental validation and results in patients with operated heart defects].

PURPOSE: A flow-sensitive MR sequence (phase-contrast technique) was evaluated in phantom studies with regard to factors influencing measurements and correctness of results. The sequence was additionally used for functional evaluation of operated congenital heart disease. METHODS: Pulsatile and constant flow were produced with the help of a phantom. Influence of angulation, range and vessel bending was evaluated. An examination protocol was developed from the results. 35 patients with surgically repaired congenital heart disease or without repair were examined. RESULTS: A range preset below the actual flow velocity as well as angulation of more than 20 degrees were isolated as main pitfalls in MR flowmetry. In addition to morphological MR findings flow measurements were possible in 11 patients at vessel sites which were not or not completely suited for examination by Doppler ultrasound. CONCLUSION: The evaluated phase-contrast technique allows for fast and reliable flow quantification if the influences identified in phantom studies are considered.

Blood Flow Velocity↗

[Experimental validation of a semi-quantitative single-layer procedure for proton-spin tomographic imaging of regional cerebral perfusion].

PURPOSE: The aim of the present study was to validate a simple MRI-procedure for semiquantitative assessment of regional cerebral blood flow. MATERIALS AND METHODS: Unilateral cerebral ischaemia (30 minutes) in the territory of the middle cerebral artery was induced in 14 anesthetised rates. The MRI-experiment consisted in an intravenous bolus injection of gadolinium-DTPA, recording of the cerebral contrast kinetics with a T2*-weighted pulse sequence, and measurement of the maximal concentration change at a chosen reference point of time. To measure perfusion quantitatively, a microsphere technique, an accepted reference technique was used. With both methods a perfusion index related to the contralateral side was calculated. RESULTS: In all cases decreased perfusion was detected by the MRI technique. The perfusion indices correlated with a coefficient of correlation of r = 0.89 (p < 0.001). CONCLUSION: The results demonstrate that contrast-enhanced MRI with bolus injection can be implemented with clinical potential as a semiquantitative instrument for the assessment of cerebral perfusion. Regional cerebral blood volume and collateral blood flow may interfere with the estimate of blood flow.

Animals↗

Computerized polymorphic marker identification: experimental validation and a predicted human polymorphism catalog.

A computational system for the prediction of polymorphic loci directly and efficiently from human genomic sequence was developed and verified. A suite of programs, collectively called POMPOUS (polymorphic marker prediction of ubiquitous simple sequences) detects tandem repeats ranging from dinucleotides up to 250 mers, scores them according to predicted level of polymorphism, and designs appropriate flanking primers for PCR amplification. This approach was validated on an approximately 750-kilobase region of human chromosome 3p21.3, involved in lung and breast carcinoma homozygous deletions. Target DNA from 36 paired B lymphoblastoid and lung cancer lines was amplified and allelotyped for 33 loci predicted by POMPOUS to be variable in repeat size. We found that among those 36 predominately Caucasian individuals 22 of the 33 (67%) predicted loci were polymorphic with an average heterozygosity of 0.42. Allele loss in this region was found in 27/36 (75%) of the tumor lines using these markers. POMPOUS provides the genetic researcher with an additional tool for the rapid and efficient identification of polymorphic markers, and through a World Wide Web site, investigators can use POMPOUS to identify polymorphic markers for their research. A catalog of 13,261 potential polymorphic markers and associated primer sets has been created from the analysis of 141,779,504 base pairs of human genomic sequence in GenBank. This data is available on our Web site (pompous.swmed.edu) and will be updated periodically as GenBank is expanded and algorithm accuracy is improved.

Computer Communication Networks↗

Modeling and experimental validation of the binary complex of the plectin actin-binding domain and the first pair of fibronectin type III (FNIII) domains of the beta4 integrin.

The binding of plectin to the beta4 subunit of the alpha6beta4 integrin is a critical step in the formation of hemidesmosomes. An important interaction between these two proteins occurs between the actin-binding domain (ABD) of plectin and the first pair of fibronectin type III (FNIII) domains and a small part of the connecting segment of beta4. Previously, a few amino acids, critical for this interaction, were identified in both plectin and beta4 and mapped on the crystal structures of the ABD of plectin and the first pair of FNIII domains of beta4. In the present study, we used this biochemical information and protein-protein docking calculations to construct a model of the binary complex between these two protein domains. The top scoring computational model predicts that the calponin-homology 1 (CH1) domain of the ABD associates with the first and the second FNIII domains of beta4. Our mutational analysis of the residues at the proposed interface of both the FNIII and the CH1 domains is in agreement with the suggested interaction model. Computational simulations to predict protein motions suggest that the exact model of FNIII and plectin CH1 interaction might well differ in detail from the suggested model due to the conformational plasticity of the FNIII domains, which might lead to a closely related but different mode of interaction with the plectin-ABD. Furthermore, we show that Ser-1325 in the connecting segment of beta4 appears to be essential for the recruitment of plectin into hemidesmosomes in vivo. This is consistent with the proposed model and previously published mutational data. In conclusion, our data support a model in which the CH1 domain of the plectin-ABD associates with the groove between the two FNIII domains of beta4.

Actins↗

Predictions of CCR1 chemokine receptor structure and BX 471 antagonist binding followed by experimental validation.

A major challenge in the application of structure-based drug design methods to proteins belonging to the superfamily of G protein-coupled receptors (GPCRs) is the paucity of structural information (1). The 19 chemokine receptors, belonging to the Class A family of GPCRs, are important drug targets not only for autoimmune diseases like multiple sclerosis but also for the blockade of human immunodeficiency virus type 1 entry (2). Using the MembStruk computational method (3), we predicted the three-dimensional structure of the human CCR1 receptor. In addition, we predicted the binding site of the small molecule CCR1 antagonist BX 471, which is currently in Phase II clinical trials (4). Based on the predicted antagonist binding site we designed 17 point mutants of CCR1 to validate the predictions. Subsequent competitive ligand binding and chemotaxis experiments with these mutants gave an excellent correlation to these predictions. In particular, we find that Tyr-113 and Tyr-114 on transmembrane domain 3 and Ile-259 on transmembrane 6 contribute significantly to the binding of BX 471. Finally, we used the predicted and validated structure of CCR1 in a virtual screening validation of the Maybridge data base, seeded with selective CCR1 antagonists. The screen identified 63% of CCR1 antagonists in the top 5% of the hits. Our results indicate that rational drug design for GPCR targets is a feasible approach.

Animals↗

Experimental validation of OT solutions to the comprehension-production dilemma.

The relationship between perception and production remains an unresolved issue within the study of phonological acquisition. Recent developments in optimality theory offer potentially new solutions to this long-standing problem; but thus far, the proposals that have been advanced are in the absence of actual perception-production data from a given child. This paper provides an empirical instantiation of the perception-production effects by appealing to data from children's knowledge and use of syllables and segments. The demonstration shows that optimality theoretic accounts of the perception-production interface hold in part, but a crucial modification is required in order to obtain the full range of effects that are observed in phonological acquisition.

Child↗

Comprehensive analysis of mRNA-microRNA-lncRNA expression profiles in post-traumatic elbow heterotopic ossification using RNA sequencing and experimental validation.

BACKGROUND: This study aimed to profile the molecular signatures of post-traumatic elbow heterotopic ossification (HO) to identify key regulators and potential therapeutic targets. METHODS: Total RNA from post-traumatic elbow HO tissues (n=4) and normal bone tissues (n=6) was subjected to high-throughput sequencing to identify differentially expressed mRNAs (DEGs), microRNAs (DEMs), and lncRNAs (DELs). Bioinformatics analyses included Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, protein-protein interaction network construction, and transcription factor (TF)-microRNA-mRNA network analysis. The expression trends of four most upregulated and four most downregulated DEGs were validated by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). RESULTS: We identified 2,138 DEGs, 40 DEMs, and 905 DELs. DEGs were significantly enriched in biological process "bone mineralization," cellular component "plasma membrane," molecular function "integrin binding," and pathways including PI3K-Akt, NF-&#x3ba;B, JAK-STAT, and TNF signaling pathways. Hub genes with high connectivity included MMP9, IL6, MMP3, CTSK, and BGLAP. Integrated network analysis highlighted the transcription factor JUN and key microRNAs (hsa-miR-124-3p, hsa-miR-548c-3p, and hsa-miR-135b). The qRT-PCR results confirmed the expression trends of selected DEGs. CONCLUSIONS: This study, for the first time, profiled the differentially expressed mRNAs, microRNAs, and lncRNAs in post-traumatic elbow HO using high-throughput RNA sequencing. These findings provide valuable insights into the molecular mechanisms of HO following elbow trauma. The identified hub genes (MMP9, IL6, MMP3, CTSK, and BGLAP), key TF (JUN), and key microRNAs (hsa-miR-124-3p, hsa-miR-548c-3p, and hsa-miR-135b) may serve as potential therapeutic targets for preventing and treating post-traumatic elbow HO.

Humans↗

Development and Experimental Validation of a Fluid/Structure-Interaction Finite Element Model of a Vacuum-Driven Cell Culture Mechanostimulus System.

A new fluid/structure-interaction finite element formulation is reported, by means of which reactive fluid stresses can be determined for what is currently the most widely used laboratory apparatus (the Flexercell Strain Unit) for delivering controlled in vitro mechanical stimuli to cultured cells. The apparatus functions by means of cyclic vacuum application to the undersurface of a membrane-like circular rubber substrate. When operated in its original embodiment (i.e., without axial constraint to substrate motion), the pulsatile vacuum causes appreciable pulsatile excursions (often several millimeters) of the substrate. The mechanical stimuli experienced by cells attached atop the substrate include not only substrate distention, but also potentially confounding reactive fluid stresses due to coupled motions of the overlying liquid culture nutrient medium. Since it is impractical to directly measure reactive fluid stress in such environments, a corresponding mathematical model has been developed. The formulation involves transient continuum finite element solutions for the nutrient medium flow field and for the deformation of the substrate, coupled at their mutual interface (the substrate culture surface). Besides the nonlinearities inherent in the flow field and substrate treatments per se, the numerical problem is complicated by the presence of moving boundaries at the nutrient free surface and at the nutrient/substrate interface, as well as by the need to enforce fluid/structure interaction throughout the duty cycle. Algorithmic considerations appropriate to achieving physically realistic numerical performance are reported, and a confirmatory laboratory validation experiment is described.

Journal Article↗

Experimental validation of potential and turbulent flow models for a two-dimensional jet enhanced exhaust hood.

A two-dimensional jet-reinforced slot exhaust hood was modeled using a mathematical model based on potential flow theory and with a computational fluid dynamics (CFD) model using the standard k-epsilon model for turbulence closure. The accuracy of the calculations was verified by air velocity and capture efficiency measurements. The comparisons show that, for normal operating conditions, both the models predicted the mean airflows in front of the hood well. However, the CFD model gave more realistic results in the jet flow region and also of the short-circuiting flow. Both models became increasingly inaccurate when the ratio of the supply jet momentum to the exhaust flow rate increased. The jet enhancement proved to be a very efficient way to increase the effective control range of exhaust hoods. Controlled air movements can be created at distances that are two to three times larger than with conventional suction alone without increasing the exhaust flow rate.

Air Pollution, Indoor↗