Mechanistic Validity Research Articles

Development and validation of clinically Mimicable model of frostbite injury-induced chronic pain

Frostbite, a debilitating condition, significantly affects the well-being of military veterans and high-altitude residents, causing severe clinical complications such as chronic pain that markedly impacts overall quality of life. There has been a notable increase in the development of pre-clinical models for studying frostbite injury, but their suitability for pain evaluation remains limited. The major hurdle in the development of novel therapeutics for the treatment of frostbite-induced chronic pain is the unavailability of well-established preclinical models. In this study, we employed deep-frozen magnets to induce frostbite injury and conducted validation for chronic pain through assessments of face, predictive, and mechanistic validity. Behavioral assays demonstrated that animals with frostbite injury exhibited significant mechanical, thermal & cold hypersensitivity in injured rats. Further, molecular analysis indicated that frostbite injury triggered the activation of TRP channels (TRPA1 and TRPM8), microgliosis, and increased neuroinflammation in the dorsal root ganglion (DRG) and spinal cord of rats. Notably, NR2B protein expressions were significantly upregulated in the DRG of injured rats, while no changes were observed in spinal NR2B expressions. Furthermore, the administration of ibuprofen (25, 50, and 100 mg/kg, i.p.) resulted in a significant improvement in behavioral, biochemical, and molecular alterations in frostbite-injured rats. Overall, results suggested that established frostbite model effectively recapitulates face, pharmacological, and mechanistic validity, highlighting its potential for screening future treatment modalities and exploring the intricate mechanisms associated with frostbite-induced chronic pain.

Translational Insights into Molecular Mechanisms of Chemical Hepatocarcinogenesis for Improved Human Risk Assessment

Background: Hepatocellular carcinoma (HCC) is a prevalent liver cancer with major risk factors being hepatitis viral infections, alcohol, non-alcoholic fatty liver disease, and aflatoxin exposure. Both genotoxic and non-genotoxic agents can induce HCC through mechanisms involving DNA damage, oxidative stress, chronic inflammation, and disrupted signaling pathways like MAPK/ERK, PI3K/AKT, WNT/β-catenin and PPARα. While rodent assays are utilized to detect potential chemical hepatocarcinogens, species differences in pathways like PPARα and CAR/PXR activation impact human risk assessment. Purpose: This analysis provides an updated, critical examination of species concordance in mechanisms of hepatic carcinogenesis to inform human safety assessment of rodent liver tumor findings. Main Body: Rodent assays including 2-year bioassays, transgenic models, and short-term studies detect liver tumors through lifetime exposure or early biomarkers. However, rodent-specific PPARα and CAR/PXR activation, along with human risk factors like hepatitis, highlight key interspecies differences. Determining mode of action relevance requires evaluating mechanistic validity and pivotal key events leading to tumors across species. Non-genotoxic compounds eliciting rodent liver tumors can activate PPARα, CAR/PXR, and other pathways triggering increased cell replication; but downstream signaling may differ in human liver. Understanding applicability of these mechanisms in humans as well as incorporating human risk factors into experimental models is critical for accurate risk assessment. Conclusion: In summary, elucidating conserved versus divergent molecular mechanisms of hepatic carcinogenesis between rodents and humans is essential for appropriately interpreting rodent findings and safeguarding human health through science-based risk assessment frameworks and regulatory decision-making processes around potential chemical hazards.

Comparative modelling results between a separable and a non-separable form of principal stretches–based strain energy functions for a variety of isotropic incompressible soft solids: Ogden model compared with a parent model

This paper presents a comparative analysis on the application of two models from the family of principal stretches–based strain energy functions to a wide range of incompressible rubber-like materials, including hydrogels, foams, silicone elastomers, filled rubbers and biomaterials. The two models of concern here are the seminal Ogden model of the separable form, and a more general non-separable form parent model proposed by the author. To this end, the three-term Ogden model, with corresponding six model parameters, and the one-term parent model with four model parameters, are applied to exemplar extant deformation datasets of the aforementioned rubber-like materials, and the ensuing modelling results are then compared. This comparison encompasses the quality of the obtained fits as measured by the resulting relative errors, and the mechanistic validity of the ensuing strain energy function as assessed by the convexity of the iso-energy plots in the principal stretches λ1,λ2\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$\\left({\\lambda }_{1},{\\lambda }_{2}\\right)$$\\end{document} plane. The studied modes of deformation involve a wide range of deformation types including uniaxial, biaxial, simple shear and inflation. It is shown that for the considered range of materials and deformations, the optimal fits obtained by the Ogden model either (i) possess higher relative errors; or (ii) offer a marginal improvement but with the loss of convexity. This loss of convexity is further exemplified on considering an additional soft tissue dataset. Finally, an important implication of the separable versus non-separable functional forms of the two models in application to the deformation of elastomers will be demonstrated, where a separable function proves to be an inappropriate choice. In all these applications, the proposed parent model, by contrast, will be shown to provide equally good or better fits, with fewer model parameters, while remaining free of the foregoing undesirable modelling outcomes.

Oxygenation of Substituted (Phenylthio)acetic Acids by Acidic Bromate: A Kinetic and Mechanistic Study and Validity of Linear Free-Energy Relationships

Kinetics and mechanistic investigations on the oxygenation of (phenylthio)acetic acid and its substituted compounds using bromate in acid medium have been carried out. The reaction exhibited first-order in [bromate], 1.6 order in [H2SO4], less than one order in [substrate] and displayed solvent isotopic effect of 1.75 (kD2O/kH2O). The reaction rate is not affected by ionic strength variation, however, enhanced by lowering dielectric constant of the medium. Structural modifications in the aryl moiety of the substrate resulted in a change of reactivity, where electron-seeking substituents decreased the reaction rate. The order of reactivity among the studied substrates is paramethoxy > para-methyl > -H > para-chloro ≈ para-bromo > para-nitro(phenylthio)acetic acid and showed an excellent correlation between rate constant and the Hammett substituent constant (σ value). The reaction constant (ρ) value is negative (-1.37 at 303 K) and decreased by increase in the reaction temperature. Proposed mechanism involves decomposition of the complex formed between acid bromate and the substrate. Based on the proposed mechanism, an appropriate rate law has been derived and tested for its validity. Activation parameters and isokinetic temperature have been evaluated and discussed. The validity of linear free-energy relationships have been discussed thoroughly.

Effects of Iodonium Analogs on Nadph Oxidase 1 in Human Colon Cancer Cells.

Recent studies suggest that of the molecules postulated to function as inhibitors of the NADPH oxidase family of enzymes iodonium analogs known to broadly interfere with flavin dehydrogenase function demonstrate mechanistic validity as NADPH oxidase poisons. In recent work, we have produced a series of novel iodonium compounds as putative inhibitors of these oxidases. To evaluate the potential utility of two novel molecules with favorable chemical properties, NSC 740104 and NSC 751140, we compared effects of these compounds to the two standard inhibitors of this class, diphenyleneiodonium and di-2-thienyliodonium, with respect to antiproliferative, cell cycle, and gene expression effects in human colon cancer cells that require the function of NADPH oxidase 1. Both new agents blocked NADPH oxidase-related reactive oxygen production, inhibited tumor cell proliferation, produced a G1/S block in cell cycle progression, and inhibited NADPH oxidase 1 expression at the mRNA and protein levels at low nM concentrations in a fashion similar to or better than the parent molecules. These studies suggest that NSC 740104 and NSC 751140 should be developed further as mechanistic tools to better understand the role of NADPH oxidase inhibition as an approach to the development of novel therapeutic agents for colon cancer.

The Neurotransmitters Involved in Drosophila Alcohol-Induced Behaviors.

Alcohol is a widely used and abused substance with numerous negative consequences for human health and safety. Historically, alcohol's widespread, non-specific neurobiological effects have made it a challenge to study in humans. Therefore, model organisms are a critical tool for unraveling the mechanisms of alcohol action and subsequent effects on behavior. Drosophila melanogaster is genetically tractable and displays a vast behavioral repertoire, making it a particularly good candidate for examining the neurobiology of alcohol responses. In addition to being experimentally amenable, Drosophila have high face and mechanistic validity: their alcohol-related behaviors are remarkably consistent with humans and other mammalian species, and they share numerous conserved neurotransmitters and signaling pathways. Flies have a long history in alcohol research, which has been enhanced in recent years by the development of tools that allow for manipulating individual Drosophila neurotransmitters. Through advancements such as the GAL4/UAS system and CRISPR/Cas9 mutagenesis, investigation of specific neurotransmitters in small subsets of neurons has become ever more achievable. In this review, we describe recent progress in understanding the contribution of seven neurotransmitters to fly behavior, focusing on their roles in alcohol response: dopamine, octopamine, tyramine, serotonin, glutamate, GABA, and acetylcholine. We chose these small-molecule neurotransmitters due to their conservation in mammals and their importance for behavior. While neurotransmitters like dopamine and octopamine have received significant research emphasis regarding their contributions to behavior, others, like glutamate, GABA, and acetylcholine, remain relatively unexplored. Here, we summarize recent genetic and behavioral findings concerning these seven neurotransmitters and their roles in the behavioral response to alcohol, highlighting the fitness of the fly as a model for human alcohol use.

Selective cytotoxic and anti-metastatic activity in DU-145 prostate cancer cells induced by Annona muricata L. bark extract and phytochemical, annonacin

BackgroundAnnona muricata L. was identified as a popular medicinal plant in treatment regimens among cancer patients in Jamaica by a previously conducted structured questionnaire. Ethnomedically used plant parts, were examined in this study against human prostate cancer cells for the first time and mechanisms of action elucidated for the most potent of them, along with the active phytochemical, annonacin.MethodsNine extracts of varying polarity from the leaves and bark of A. muricata were assessed initially for cytotoxicity using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay on PC-3 prostate cancer cells and the ethyl acetate bark (EAB) extract was identified as the most potent. EAB extract was then standardized for annonacin content using High-performance Liquid Chromatography - Mass Spectrometry (HPLC-MS) and shown to be effective against a second prostate cancer cell line (DU-145) also. The mode of cell death in DU-145 cells were assessed via several apoptotic assays including induction of increased reactive oxygen species (ROS) production, reduction of mitochondrial membrane potential, activation of caspases and annexin V externalization combined with morphological observations using confocal microscopy. In addition, the potential to prevent metastasis was examined via inhibition of cell migration, vascular endothelial growth factor (VEGF) and angiogenesis using the chorioallantoic membrane assay (CAM).ResultsAnnonacin and EAB extract displayed selective and potent cytotoxicity against the DU-145 prostate carcinoma cells with IC50 values of 0.1 ± 0.07 μM and 55.501 ± 0.55 μg/mL respectively, without impacting RWPE-1 normal prostate cells, in stark contrast to chemotherapeutic docetaxel which lacked such selectivity. Docetaxel’s impact on the cancerous DU-145 was improved by 50% when used in combination with EAB extract. Insignificant levels of intracellular ROS content, depolarization of mitochondrial membrane, Caspase 3/7 activation, annexin V content, along with stained morphological evaluations, pointed to a non-apoptotic mode of cell death. The extract at 50 μg/mL deterred cell migration in the wound-healing assay, while inhibition of angiogenesis was displayed in the CAM and VEGF inhibition assays for both EAB (100 μg /mL) and annonacin (0.5 μM).ConclusionsTaken together, the standardized EAB extract and annonacin appear to induce selective and potent cell death via a necrotic pathway in DU-145 cells, while also preventing cell migration and angiogenesis, which warrant further examinations for mechanistic insights and validity in-vivo.

Flying Together: Drosophila as a Tool to Understand the Genetics of Human Alcoholism.

Alcohol use disorder (AUD) exacts an immense toll on individuals, families, and society. Genetic factors determine up to 60% of an individual’s risk of developing problematic alcohol habits. Effective AUD prevention and treatment requires knowledge of the genes that predispose people to alcoholism, play a role in alcohol responses, and/or contribute to the development of addiction. As a highly tractable and translatable genetic and behavioral model organism, Drosophila melanogaster has proven valuable to uncover important genes and mechanistic pathways that have obvious orthologs in humans and that help explain the complexities of addiction. Vinegar flies exhibit remarkably strong face and mechanistic validity as a model for AUDs, permitting many advancements in the quest to understand human genetic involvement in this disease. These advancements occur via approaches that essentially fall into one of two categories: (1) discovering candidate genes via human genome-wide association studies (GWAS), transcriptomics on post-mortem tissue from AUD patients, or relevant physiological connections, then using reverse genetics in flies to validate candidate genes’ roles and investigate their molecular function in the context of alcohol. (2) Utilizing flies to discover candidate genes through unbiased screens, GWAS, quantitative trait locus analyses, transcriptomics, or single-gene studies, then validating their translational role in human genetic surveys. In this review, we highlight the utility of Drosophila as a model for alcoholism by surveying recent advances in our understanding of human AUDs that resulted from these various approaches. We summarize the genes that are conserved in alcohol-related function between humans and flies. We also provide insight into some advantages and limitations of these approaches. Overall, this review demonstrates how Drosophila have and can be used to answer important genetic questions about alcohol addiction.

Genomic risk prediction of aromatase inhibitor-related arthralgia in patients with breast cancer using a novel machine-learning algorithm.

Many breast cancer (BC) patients treated with aromatase inhibitors (AIs) develop aromatase inhibitor‐related arthralgia (AIA). Candidate gene studies to identify AIA risk are limited in scope. We evaluated the potential of a novel analytic algorithm (NAA) to predict AIA using germline single nucleotide polymorphisms (SNP) data obtained before treatment initiation. Systematic chart review of 700 AI‐treated patients with stage I‐III BC identified asymptomatic patients (n = 39) and those with clinically significant AIA resulting in AI termination or therapy switch (n = 123). Germline DNA was obtained and SNP genotyping performed using the Affymetrix UK BioBank Axiom Array to yield 695,277 SNPs. SNP clusters that most closely defined AIA risk were discovered using an NAA that sequentially combined statistical filtering and a machine‐learning algorithm. NCBI PhenGenI and Ensemble databases defined gene attribution of the most discriminating SNPs. Phenotype, pathway, and ontologic analyses assessed functional and mechanistic validity. Demographics were similar in cases and controls. A cluster of 70 SNPs, correlating to 57 genes, was identified. This SNP group predicted AIA occurrence with a maximum accuracy of 75.93%. Strong associations with arthralgia, breast cancer, and estrogen phenotypes were seen in 19/57 genes (33%) and were functionally consistent. Using a NAA, we identified a 70 SNP cluster that predicted AIA risk with fair accuracy. Phenotype, functional, and pathway analysis of attributed genes was consistent with clinical phenotypes. This study is the first to link a specific SNP/gene cluster to AIA risk independent of candidate gene bias.

Genomic risk prediction of aromatase inhibitor-related arthralgias (AIA) in breast cancer (BC) patients using a novel analytical algorithm (NAA).

10102 Background: Many BC patients treated with aromatase inhibitors (AIs) develop AIA; 20% have symptoms severe enough to effect treatment compliance. Results of candidate gene studies to identify AIA risk are limited in scope. In this case-controlled study, we evaluated the potential of a NAA to predict AIA using germline single nucleotide polymorphism (SNP) data obtained prior to treatment initiation. Methods: Systematic chart review of 700 AI-treated patients with stage I-III BC between 2003-2012 identified asymptomatic patients (n = 39) and those with clinically significant AIA resulting in AI termination or therapy switch (n = 123). Germline DNA was obtained from peripheral blood cells and SNP genotyping performed using the Affymetrix UK BioBank Axiom Array to yield 695,277 SNPs. The identity of the cluster of SNPs that most closely defined AIA risk was discovered using an NAA that sequentially combined statistical filtering and a machine learning algorithm. NCBI PhenGenI and Ensemble databases were used to define gene attribution of the 200 most discriminating SNPs. Phenotype, pathway, and ontologic analyses assessed functional and mechanistic validity. Results: Cases and controls were similar in demographic characteristics. A cluster of 70 SNPs, correlated to 57 genes (accounting for linkage disequilibrium), was identified. This SNP group predicted AIA occurrence with a maximum accuracy of 75.93%. Strong associations with arthralgia, breast cancer, and estrogen phenotypes were seen in 19/57 genes (33%) and were functionally and ontologically consistent. Conclusions: Using a NAA, we identified a 70 SNP cluster that predicted AIA risk with fair accuracy. Phenotype, functional, and pathway analysis of attributed genes was consistent with clinical phenotypes. This study is the first to link a specific SNP/gene cluster to AIA risk independent of candidate gene bias. An ongoing prospective companion study will be used to validate and to expand upon results.

Abstract A15: HDAC6 inhibitor C1A abrogates the recruitment of the autophagic machinery and synergizes with proteasome, src kinase, and PI3K-mTOR inhibition.

Abstract The histone deacetylase (HDAC) family of proteins plays a profound role in transcriptional silencing and intracellular protein function, and HDAC inhibitors have shown promise for the treatment of human malignancies. HDAC6 activity is altered in tumorigenesis and its selective inhibition is likely to cause less toxicity compared to broad spectrum inhibitors. An important function of HDAC6 is the processing of misfolded proteins within cells during autophagy and formation of aggresomes. Prosurvival autophagy may represent a major impediment to successful cancer therapy and more potent and specific inhibitors of autophagy are needed. This study evaluated whether targeting HDAC6 could be used to impair autophagy and restore chemosensitivity. Treatment of colon cancer HCT-116 cells and osteosarcoma U2OS-LC3-GFP cells with HDAC6 inhibitor C1A for 24 hours (or tubastatin A, another HDAC6 inhibitor) caused a dose dependent increase in expression of lipid associated LC3 II, suggesting autophagosome formation or blockage. Using a pH-sensitive, double tagged mCherry-GFP-LC3 reporter, we could further demonstrate impairment of autophagosome-lysosome fusion. We hypothesized that the effect of autophagy tool compounds will be modulated by C1A. Increased LC3 II/p62 expression induced by chloroquine, 3-MA, bortezomib or BEZ-235 were inhibited by C1A (also seen with tubastatin A). Combination of C1A with bortezomib or BEZ-235 induced a greater proportion of cells undergoing apoptosis as depicted by an increase of caspase 3/7 activity in HCT-116 cells. Treatment with C1A (or tubastatin A) also impaired the formation of LC3 II following treatment with src inhibitor, dasatinib (or PP2, another src inhibitor but not PP3, a negative control compound); synergy as assessed by caspase 3/7 activity was seen in lung cancer A549 cells. Based on combination index values, we demonstrated that C1A synergistically enhanced proteasome, src kinase and PI3K-mTOR growth inhibition. In vivo, the anti-tumor activity of C1A in combination with bortezomib or BEZ-235 was confirmed in HCT-116 tumor bearing mice. Tumor growth was significantly delayed in animals treated with combination therapy. A similar observation was seen in A549 tumor bearing mice treated with C1A in combination with dasatinib. The efficacy of the combination C1A and BEZ-235 was also predicted as early as 48 h with [18F]fluorothymidine positron emission tomography ([18F]FLT-PET) in HCT-116 tumor bearing mice. These results provide a rationale for combination therapy with HDAC6 inhibitors and chemotherapy associated with pro-survival autophagy in the treatment of solid tumors. Imaging of cell proliferation with [18F]FLT-PET detected the mechanistic validity of this combination treatment approach and, thus, warrants further investigation in pre-clinical models and patients. This work was supported in part by Cancer Research U.K.-Engineering and Physical Sciences Research Council Grant C2536/A10337. Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A15. Citation Format: Maciej Kaliszczak, Olivier E. Pardo, Michael J. Seckl, Eric O. Aboagye. HDAC6 inhibitor C1A abrogates the recruitment of the autophagic machinery and synergizes with proteasome, src kinase, and PI3K-mTOR inhibition. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A15.

Extending, changing, and explaining the brain

This paper addresses concerns raised recently by Datteri (Biol Philos 24:301–324, 2009) and Craver (Philos Sci 77(5):840–851, 2010) about the use of brain-extending prosthetics in experimental neuroscience. Since the operation of the implant induces plastic changes in neural circuits, it is reasonable to worry that operational knowledge of the hybrid system will not be an accurate basis for generalisation when modelling the unextended brain. I argue, however, that Datteri’s no-plasticity constraint unwittingly rules out numerous experimental paradigms in behavioural and systems neuroscience which also elicit neural plasticity. Furthermore, I propose that Datteri and Craver’s arguments concerning the limitations of prosthetic modelling in basic neuroscience, as opposed to neuroengineering, rests on too narrow a view of the ways models in neuroscience should be evaluated, and that a more pluralist approach is needed. I distinguish organisational validity of models from mechanistic validity. I argue that while prosthetic models may be deficient in the latter of these explanatory virtues because of neuroplasticity, they excel in the former since organisational validity tracks the extent to which a model captures coding principles that are invariant with plasticity. Changing the brain, I conclude, is one viable route towards explaining the brain.

Criteria of validity for animal models of psychiatric disorders: focus on anxiety disorders and depression

Animal models of psychiatric disorders are usually discussed with regard to three criteria first elaborated by Willner; face, predictive and construct validity. Here, we draw the history of these concepts and then try to redraw and refine these criteria, using the framework of the diathesis model of depression that has been proposed by several authors. We thus propose a set of five major criteria (with sub-categories for some of them); homological validity (including species validity and strain validity), pathogenic validity (including ontopathogenic validity and triggering validity), mechanistic validity, face validity (including ethological and biomarker validity) and predictive validity (including induction and remission validity). Homological validity requires that an adequate species and strain be chosen: considering species validity, primates will be considered to have a higher score than drosophila, and considering strains, a high stress reactivity in a strain scores higher than a low stress reactivity in another strain. Pathological validity corresponds to the fact that, in order to shape pathological characteristics, the organism has been manipulated both during the developmental period (for example, maternal separation: ontopathogenic validity) and during adulthood (for example, stress: triggering validity). Mechanistic validity corresponds to the fact that the cognitive (for example, cognitive bias) or biological mechanisms (such as dysfunction of the hormonal stress axis regulation) underlying the disorder are identical in both humans and animals. Face validity corresponds to the observable behavioral (ethological validity) or biological (biomarker validity) outcomes: for example anhedonic behavior (ethological validity) or elevated corticosterone (biomarker validity). Finally, predictive validity corresponds to the identity of the relationship between the triggering factor and the outcome (induction validity) and between the effects of the treatments on the two organisms (remission validity). The relevance of this framework is then discussed regarding various animal models of depression.

Aquachlororuthenium(III) catalysis in the oxidation of substituted 4-oxo-4-arylbutanoic acids by bromate in acid medium: a kinetic and mechanistic study and validity of linear free-energy relationships

Ru(III) acts an efficient catalyst in the oxidation of substituted 4-oxo-4-arylbutanoic acids (4-oxo acids) by bromate in sulfuric acid medium, giving the corresponding benzoic acids in quantitative yields. The reaction shows first-order dependence in both [bromate] and [H2SO4], and a non-linear dependence on both [oxo acid] and [catalyst]. Changing solvent from H2O to D2O increases the rate. The rate is not affected by ionic strength but decreases with increase in dielectric constant of the medium. Electron-releasing substituents in the phenyl ring of the substrate greatly accelerate the rate, whereas the retardation by electron-withdrawing substituents, though perceptible, is small. The linear free-energy relationship is characterized by smooth curves in Hammett plots of log k versus σ; however, linear plots are obtained with excellent correlation coefficients at all the studied temperatures, when Brown’s σ+ values are used. The reaction constant is negative and decreases with increase in temperature. From the intersection of the lines in the Hammett and Arrhenius plots, the isokinetic relationship is evaluated. A mechanism involving a cyclic oxidant–substrate–catalyst ternary complex is proposed, in which both C–C bond-breaking and C–O bond formation are involved, and the oxidation state of Ru(III) remains unchanged. A rate law explaining all the kinetic results has been derived and verified. The reaction is an example of neighboring group participation in intramolecular catalysis and is potentially useful for the synthesis of substituted benzoic acids.

A Mechanistic Model of Material Removal in Magnetorheological Finishing (MRF)

According to the sharp rheological characteristics of magnetorheological fluid in the magnetic field, the principle and mechanism of magnetorheological finishing is analyzed. Based on the Preston equation, the Reynolds equation and its boundary conditions, the two-dimensional material removal model is built and simulated. Furthermore, a series of MRF experiments are carried out, and the influence of the immersed depth and material kinds on material removal rate are clarified respectively. The experimental results are compared with the modeling results of material removal rate to confirm the mechanistic model validity.

Modulation of synchrony without changes in firing rates.

It was often reported and suggested that the synchronization of spikes can occur without changes in the firing rate. However, few theoretical studies have tested its mechanistic validity. In the present study, we investigate whether changes in synaptic weights can induce an independent modulation of synchrony while the firing rate remains constant. We study this question at the level of both single neurons and neuronal populations using network simulations of conductance based integrate-and-fire neurons. The network consists of a single layer that includes local excitatory and inhibitory recurrent connections, as well as long-range excitatory projections targeting both classes of neurons. Each neuron in the network receives external input consisting of uncorrelated Poisson spike trains. We find that increasing this external input leads to a linear increase of activity in the network, as well as an increase in the peak frequency of oscillation. In contrast, balanced changes of the synaptic weight of excitatory long-range projections for both classes of postsynaptic neurons modulate the degree of synchronization without altering the firing rate. These results demonstrate that, in a simple network, synchronization and firing rate can be modulated independently, and thus, may be used as independent coding dimensions.

Synaptic Cysteine Sulfhydryl Groups as Targets of Electrophilic Neurotoxicants

Many structurally diverse chemicals (e.g., acrylamide, 2,4-dithiobiuret, methylmercury) are electrophiles and cause synaptic dysfunction by unknown mechanisms. The purpose of this Forum review is to discuss the possibility that highly nucleophilic cysteine thiolate groups within catalytic triads of synaptic proteins represent specific and necessary targets for electrophilic neurotoxicants. Most of these toxicants share the ability to adduct or otherwise modify nucleophilic sulfhydryl groups. It is also now recognized that synaptic activity is regulated by the redox state of certain cysteine sulfhydryl groups on proteins. Electrophilic neurotoxicants might, therefore, produce synaptic toxicity by modifying these thiols. Because most proteins contain cysteine residues, target specificity is an issue that significantly detracts from the mechanistic validity of this hypothesis. However, recent research indicates that these thiolates are receptors for the endogenous nitric oxide (NO) pathway and that subsequent reversible S-nitrosylation finely regulates a broad spectrum of synaptic activities. We hypothesize that electrophilic neurotoxicants selectively adduct/derivatize NO-receptor thiolates in catalytic triads and that the resulting loss of fine gain control impairs neurotransmission and produces neurotoxicity. This proposal has mechanistic implications for a large class of electrophilic chemicals used in the agricultural and industrial sectors. In addition, research based on this hypothesis could provide mechanistic insight into neurodegenerative conditions such as Parkinsonism and Alzheimer's disease that presumably involve endogenous production of neurotoxic electrophiles (e.g., acrolein, 4-hydroxy-2-nonenal). The proposed mechanism of electrophilic neurotoxicants represents a new and exciting experimental framework for mechanistic research in human neuropathological conditions associated with toxicant exposure or disease-based processes.

Performance Modeling of the Ballard Mark IV Solid Polymer Electrolyte Fuel Cell: II . Empirical Model Development

A parametric model predicting the performance of a solid polymer electrolyte, proton exchange membrane (PEM) fuel cell has been developed using a combination of mechanistic and empirical modeling techniques. This paper details the empirical analysis which yielded the parametric coefficients employed in the model. A 28 run experiment covering a range of operating currents (50 to 300 ASF), temperatures (328 to 358 K), oxygen partial pressures (0.6 to 3.1 atm abs.) and hydrogen partial pressures (2.0 to 3.1 atm abs.) was conducted. Parametric equations for the activation overvoltage and the internal resistance of the fuel cell were obtained from linear regression. The factors to be employed in the linear regression had been previously determined through a mechanistic analysis of fuel cell processes. Activation overvoltage was modeled as a function of the operating temperature, the product of operating temperature, and the logarithm of the operating current, and the product of operating temperature and the logarithm of the oxygen concentration at the catalyst reaction sites. The internal resistance of the fuel cell was modeled as a function of the operating temperature and the current. Correlation of the empirical model to experimental data was very good. It is anticipated that the mechanistic validity yielded by the coupling of mechanistic and empirical modeling techniques will also allow for accurate predictive capabilities outside of the experimental range.