Heterogeneous linear elastic trabecular bone modelling using micro-CT attenuation data and experimentally measured heterogeneous tissue properties

High-resolution voxel-based finite element software, such as FEEBE developed at the NCBES, is widely used for studying trabecular bone at the micro-scale. A new approach to determine heterogeneous bone tissue material properties for computational models was proposed in this study. The specimen-specific range of tissue moduli across strut width was determined from nanoindentation testing. This range was mapped directly using linear interpolation to that specimen's micro-computed tomography (microCT) grey value range as input material properties for finite element analysis. The method was applied to cuboid trabecular bone samples taken from eight, 4-year-old (skeletally mature) ovine L5 vertebrae. Before undergoing experimental uniaxial compression tests, the samples were microCT scanned and 30 microm resolution finite element models were generated. The linear elastic finite element models were compressed to 1% strain. This material property assignment method for computational models accurately reproduced the experimentally determined apparent modulus and concentrations of stress at locations of failure.     

Pharmacological characterization of a small molecule inhibitor of c-Jun kinase

c-Jun NH(2)-terminal kinase (JNK) plays an important role in insulin resistance; however, identification of pharmacologically potent and selective small molecule JNK inhibitors has been limited. Compound A has a cell IC(50) of 102 nM and is at least 100-fold selective against related kinases and 27-fold selective against glycogen synthase kinase-3beta and cyclin-dependent kinase-2. In C57BL/6 mice, compound A reduced LPS-mediated increases in both plasma cytokine levels and phosphorylated c-Jun in adipose tissue. Treatment of mice fed a high-fat diet with compound A for 3 wk resulted in a 13.1 +/- 1% decrease in body weight and a 9.3 +/- 1.5% decrease in body fat, compared with a 6.6 +/- 2.1% increase in body weight and a 6.7 +/- 2.1% increase in body fat in vehicle-treated mice. Mice pair fed to those that received compound A exhibited a body weight decrease of 7 +/- 1% and a decrease in body fat of 1.6 +/- 1.3%, suggesting that reductions in food intake could not account solely for the reductions in adiposity observed. Compound A dosed at 30 mg/kg for 13 days in high-fat fed mice resulted in a significant decrease in phosphorylated c-Jun in adipose tissue accompanied by a decrease in weight and reductions in glucose and triglycerides and increases in insulin sensitivity to levels comparable with those in lean control mice. The ability of compound A to reduce the insulin-stimulated phosphorylation of insulin receptor substrate-1 (IRS-1) von Ser307 and partially reverse the free fatty acid inhibition of glucose uptake in 3T3L1 adipocytes, suggests that enhancement of insulin signaling in addition to weight loss may contribute to the effects of compound A on insulin sensitization in vivo. Pharmacological inhibition of JNK using compound A may therefore offer an effective therapy for type 2 diabetes mediated at least in part via weight reduction.     

Identification of functional intramuscular rectal mechanoreceptors in aganglionic rectal smooth muscle from piebald lethal mice

The mechanosensitive endings of low-threshold, slowly adapting pelvic afferents that innervate the rectum have been previously identified as rectal intraganglionic laminar endings (rIGLEs) that lie within myenteric ganglia. We tested whether the aganglionic rectum of piebald-lethal (s(l)/s(l)) mice lacks rIGLEs and whether this could explain impaired distension-evoked reflexes from this region. Extracellular recordings were made from fine rectal nerves in C57BL/6 wild-type and s(l)/s(l) mice, combined with anterograde labeling. In C57BL/6 mice, graded circumferential stretch applied to the rectum activated graded increases in firing of slowly adapting rectal mechanoreceptors. In s(l)/s(l) mice, graded stretch of the aganglionic rectum activated similar graded increases in rectal afferent firing. Stretch-sensitive afferents responded at low mechanical thresholds and fired more intensely at noxious levels of stretch. They could also be activated by probing their receptive fields with von Frey hairs and by muscle contraction. Anterograde labeling from recorded rectal nerves identified the mechanoreceptors of muscular afferents in the aganglionic rectal smooth muscle. A population of afferents were also recorded in both C57BL/6 and s(l)/s(l) mice that were activated by von Frey hair probing, but not stretch. In summary, the aganglionic rectum is innervated by a population of stretch-sensitive rectal afferent mechanoreceptor which develops and functions in the absence of any enteric ganglia. These results suggest that in patients with Hirschsprung's disease the inability to activate extrinsic distension reflexes from the aganglionic rectum is unlikely to be due to the absence of stretch-sensitive extrinsic mechanoreceptors.     

Practical review of self-mutilation in horses

Self-mutilation in horses includes biting, stomping and kicking, rubbing, and lunging into objects. Based on the author's clinical experience, three distinct types of self-mutilation are proposed and described. Type I represents normal behavioral response to continuous or intermittent physical discomfort. Type II, seen in stallions and geldings, can be recognized as self-directed intermale aggression. The behavior includes the elements and order of the natural interactive sequence typical of encounters between two stallions, except that the stallion himself is the target of his intermale behavior. Type III involves a more quiet, often rhythmically repetitive or methodical behavioral sequence of a stereotypy, for example nipping at various areas of the body in a relatively invariant pattern, stomping, or kicking rhythmically against an object. The prevalence of the various types of self-mutilation in horses is not known. In one survey, results suggested that self-mutilation of one form or another has been observed in as many as 2% of domestic stallions. Among and between stallions, self-mutilation varies in frequency and intensity, and can reach levels that are fertility and/or life threatening. Careful evaluation of the horse's behavior is often necessary to distinguish the specific type. Type I self-mutilation, where physical discomfort is the root cause, can be eliminated by relieving the discomfort. For Types II and III, understanding of intermale interactive behavior of horses and the environmental factors that may trigger or exacerbate the self-mutilative form, can be useful in guiding humane management or behavior modification. Pharmacologic interventions may be a useful adjunct to management and nutritional changes.     

Nitrogen solutes in an Adirondack forested watershed: Importance of dissolved organic nitrogen

Nitrogen (N) dynamics were evaluated from 1 June 1995 through 31 May 1996 within the Arbutus Lake watershed in the Adirondack Mountains of New York State, USA. At the Arbutus Lake outlet dissolved organic nitrogen (DON), NO₃ ^- and NH₄ ⁺ contributed 61%, 33%, and 6% respectively, to the total dissolved nitrogen (TDN) flux (259 mol ha^-1 yr^-1). At the lake inlet DON, NO₃ ^-, and NH₄ ^- constituted 36%, 61%, and 3% respectively, of TDN flux (349 mol ha^-1 yr^-1). Differences between the factors that control DON, NO₃ ⁺, and NH₄ ⁺ stream water concentrations were evaluated using two methods for estimating annual N flux at the lake inlet. Using biweekly sampling NO₃ ^- and NH₄ ⁺ flux was 10 and 4 mol ha^-1 yr^-1 respectively, less than flux estimates using biweekly plus storm and snowmelt sampling. DON flux was 18 mol ha^-1 yr^-1 greater using only biweekly sampling. These differences are probably not of ecological significance relative to the total flux of N from the watershed (349 mol ha^-1 yr^-1). Dissolved organic N concentrations were positively related to discharge during both the dormant (R² = 0.31; P < 0.01) and growing season (R² = 0.09; P < 0.01). There was no significant relationship between NO₃ ^- concentration and discharge during the dormant season, but a significant negative relationship was found during the growing season (R² = 0.29; P < 0.01). Biotic controls in the growing season appeared to have had a larger impact on stream water NO₃ ^- concentrations than on DON concentrations. Arbutus Lake had a major impact on stream water N concentrations of the four landscape positions sampled, suggesting the need to quantify within lake processes to interpret N solute losses and patterns in watershed-lake systems.     

Multi-factorial analysis of class prediction error: estimating optimal number of biomarkers for various classification rules

Machine learning and statistical model based classifiers have increasingly been used with more complex and high dimensional biological data obtained from high-throughput technologies. Understanding the impact of various factors associated with large and complex microarray datasets on the predictive performance of classifiers is computationally intensive, under investigated, yet vital in determining the optimal number of biomarkers for various classification purposes aimed towards improved detection, diagnosis, and therapeutic monitoring of diseases. We investigate the impact of microarray based data characteristics on the predictive performance for various classification rules using simulation studies. Our investigation using Random Forest, Support Vector Machines, Linear Discriminant Analysis and k-Nearest Neighbour shows that the predictive performance of classifiers is strongly influenced by training set size, biological and technical variability, replication, fold change and correlation between biomarkers. Optimal number of biomarkers for a classification problem should therefore be estimated taking account of the impact of all these factors. A database of average generalization errors is built for various combinations of these factors. The database of generalization errors can be used for estimating the optimal number of biomarkers for given levels of predictive accuracy as a function of these factors. Examples show that curves from actual biological data resemble that of simulated data with corresponding levels of data characteristics. An R package optBiomarker implementing the method is freely available for academic use from the Comprehensive R Archive Network (http://www.cran.r-project.org/web/packages/optBiomarker/).     

The application of neoglycopeptides in the development of sensitive surface plasmon resonance-based biosensors

The development of a biosensor based on surface plasmon resonance is described for the detection of carbohydrate-binding proteins in solution on a Biacore 2000 instrument, using immobilized glycopeptides as ligands. Their selection was based on previous screenings of solid-phase glycopeptide libraries with Ricinus communis agglutinin (RCA(120)) and human adhesion/growth-regulatory galectin-1 (h-Gal-1). Glycopeptides were immobilized on Au sensor chips functionalized with mixed self-assembled monolayers of different ratios of 11-mercapto-1-undecanol and 11-mercaptoundecanoic acid, and of 3-mercapto-1-propanol and 11-mercaptoundecanoic acid. The biosensors were optimized for the detection of RCA(120), and a detection limit of 0.13nM was obtained. Subsequent experiments with h-Gal-1 indicated a detection limit of at least 0.9nM for this lectin. Additionally, the effect of interfering proteins on the sensitivity of the optimized biosensor was investigated.     

The habitat hectares approach to vegetation assessment: An evaluation and suggestions for improvement

Summary The habitat hectares approach is an explicit, quantitative method for assessing the quality of vegetation by adding scores that are assigned to 10 habitat attributes. We believe it will be more repeatable and transparent than other methods that rely on subjective judgement. However, we have four principal criticisms of the method as it is currently proposed: (i) measurement of some of the attributes may be subject to considerable error that varies among assessors; (ii) the comparison of each measure with a single benchmark does not accommodate appropriate disturbance regimes; (iii) the proposed combination of attributes leads to some apparent internal inconsistencies; and (iv) it is not clear how the method will actually be used in practice. We suggest modifications to address these concerns and improve the proposed method. Finally, we make additional suggestions about the method's potential application, including: separate reporting of the extent and quality of different vegetation types to avoid the inappropriate combination of measures of area and quality; valuing appropriate disturbance regimes in natural areas; and considering very carefully the application of compensation or mitigation measures.     

N-pyridin-3-yl- and N-quinolin-3-yl-benzamides: modulators of human vanilloid receptor 1 (TRPV1)

High throughput screening of our compound library revealed a series of N-pyridyl-3-benzamides as low micromolar agonists of the human TRPV1 receptor. Synthesis of analogs in this series led to the discovery of a series of N-quinolin-3-yl-benzamides as low nanomolar antagonists of human TRPV1.