Glucagon Fibril Polymorphism Reflects Differences in Protofilament Backbone Structure

Amyloid fibrils formed by the 29-residue peptide hormone glucagon at different concentrations have strikingly different morphologies when observed by transmission electron microscopy. Fibrils formed at low concentration (0.25 mg/mL) consist of two or more protofilaments with a regular twist, while fibrils at high concentration (8 mg/mL) consist of two straight protofilaments. Here, we explore the structural differences underlying glucagon polymorphism using proteolytic degradation, linear and circular dichroism, Fourier transform infrared spectroscopy (FTIR), and X-ray fiber diffraction. Morphological differences are perpetuated at all structural levels, indicating that the two fibril classes differ in terms of protofilament backbone regions, secondary structure, chromophore alignment along the fibril axis, and fibril superstructure. Straight fibrils show a conventional β-sheet-rich far-UV circular dichroism spectrum whereas that of twisted fibrils is dominated by contributions from β-turns. Fourier transform infrared spectroscopy confirms this and also indicates a more dense backbone with weaker hydrogen bonding for the twisted morphology. According to linear dichroism, the secondary structural elements and the aromatic side chains in the straight fibrils are more highly ordered with respect to the alignment axis than the twisted fibrils. A series of highly periodical reflections in the diffractogram of the straight fibrils can be fitted to the diffraction pattern expected from a cylinder. Thus, the highly integrated structural organization in the straight fibril leads to a compact and highly uniform fibril with a well-defined edge. Prolonged proteolytic digestion confirmed that the straight fibrils are very compact and stable, while parts of the twisted fibril backbone are much more readily degraded. Differences in the digest patterns of the two morphologies correlate with predictions from two algorithms, suggesting that the polymorphism is inherent in the glucagon sequence. Glucagon provides a striking illustration of how the same short sequence can be folded into two remarkably different fibrillar structures.     

Integrated Microbial Survey Analysis of Prokaryotic Communities for the PhyloChip Microarray

PhyloTrac is an integrated desktop application for analysis of PhyloChip microarray data. PhyloTrac combined with PhyloChip provides turnkey and comprehensive identification and analysis of bacterial and archaeal communities in complex environmental samples. PhyloTrac is free for noncommercial organizations and is available for all major operating systems at http://www.phylotrac.org/.The PhyloChip is a low-cost Affymetrix GeneChip microarray, developed at Lawrence Berkeley National Laboratory (LBNL), designed to detect and quantify abundance of bacterial and archaeal taxa using signature probes targeting all known 16S rRNA gene sequences. The second generation of the PhyloChip microarray targets nearly 9,000 operational taxonomic units (OTUs), with an average of 24 probes, each 25 bp long, and the upcoming third-generation PhyloChip application will target an even larger number of OTUs. Multiple, complex environments have been successfully analyzed using the PhyloChip microarray, including, among others, air (2), soil (1), the human lung (6), and the gut (9). PhyloChip microarrays are manufactured by Affymetrix, but to date, analysis has been available only from within LBNL, limiting the accessibility of the technology. PhyloTrac addresses this limitation by providing a standardized analysis package for the PhyloChip microarray, including microarray normalization, OTU quantification, multiple interactive visualizations, and integrated analytics.     

The use of EMG biofeedback for learning of selective activation of intra-muscular parts within the serratus anterior muscle: A novel approach for rehabilitation of scapular muscle imbalance

Motor control and learning possibilities of scapular muscles are of clinical interest for restoring scapular muscle balance in patients with neck and shoulder disorders. The aim of the study was to investigate whether selective voluntary activation of intra-muscular parts within the serratus anterior can be learned with electromyographical (EMG) biofeedback, and whether the lower serratus anterior and the lower trapezius muscle comprise the lower scapula rotation force couple by synergistic activation. Nine healthy males practiced selective activation of intra-muscular parts within the serratus anterior with visual EMG biofeedback, while the activity of four parts of the serratus anterior and four parts of the trapezius muscle was recorded. One subject was able to selectively activate both the upper and the lower serratus anterior respectively. Moreover, three subjects managed to selectively activate the lower serratus anterior, and two subjects learned to selectively activate the upper serratus anterior. During selective activation of the lower serratus anterior, the activity of this muscle part was 14.4 ± 10.3 times higher than the upper serratus anterior activity (P < 0.05). The corresponding ratio for selective upper serratus vs. lower serratus anterior activity was 6.4 ± 1.7 (P < 0.05). Moreover, selective activation of the lower parts of the serratus anterior evoked 7.7 ± 8.5 times higher synergistic activity of the lower trapezius compared with the upper trapezius (P < 0.05). The learning of complete selective activation of both the lower and the upper serratus anterior of one subject, and selective activation of either the upper or lower serratus anterior by five subjects designates the promising clinical application of EMG biofeedback for restoring scapular muscle balance. The synergistic activation between the lower serratus anterior and the lower trapezius muscle was observed in only a few subjects, and future studies including more subjects are required before conclusions of a lower scapula rotation couple can be drawn.     

Exercise increases interleukin-10 levels both intraarticularly and peri-synovially in patients with knee osteoarthritis: a randomized controlled trial

Introduction  

The microdialysis method was applied to the human knee joint with osteoarthritis (OA) in order to reveal changes in biochemical markers of cartilage and inflammation, intraarticularly and in the synovium, in response to a single bout of mechanical joint loading.     

A musculoskeletal foot model for clinical gait analysis

Several full body musculoskeletal models have been developed for research applications and these models may potentially be developed into useful clinical tools to assess gait pathologies. Existing full-body musculoskeletal models treat the foot as a single segment and ignore the motions of the intrinsic joints of the foot. This assumption limits the use of such models in clinical cases with significant foot deformities. Therefore, a three-segment musculoskeletal model of the foot was developed to match the segmentation of a recently developed multi-segment kinematic foot model. All the muscles and ligaments of the foot spanning the modeled joints were included. Muscle pathways were adjusted with an optimization routine to minimize the difference between the muscle flexion–extension moment arms from the model and moment arms reported in literature. The model was driven by walking data from five normal pediatric subjects (aged 10.6±1.57 years) and muscle forces and activation levels required to produce joint motions were calculated using an inverse dynamic analysis approach. Due to the close proximity of markers on the foot, small marker placement error during motion data collection may lead to significant differences in musculoskeletal model outcomes. Therefore, an optimization routine was developed to enforce joint constraints, optimally scale each segment length and adjust marker positions. To evaluate the model outcomes, the muscle activation patterns during walking were compared with electromyography (EMG) activation patterns reported in the literature. Model-generated muscle activation patterns were observed to be similar to the EMG activation patterns.     

Dipetalodipin, a novel multifunctional salivary lipocalin that inhibits platelet aggregation, vasoconstriction, and angiogenesis through unique binding specificity for TXA2, PGF2alpha, and 15(S)-HETE

Dipetalodipin (DPTL) is an 18 kDa protein cloned from salivary glands of the triatomine Dipetalogaster maxima. DPTL belongs to the lipocalin superfamily and has strong sequence similarity to pallidipin, a salivary inhibitor of collagen-induced platelet aggregation. DPTL expressed in Escherichia coli was found to inhibit platelet aggregation by collagen, U-46619, or arachidonic acid without affecting aggregation induced by ADP, convulxin, PMA, and ristocetin. An assay based on incubation of DPTL with small molecules (e.g. prostanoids, leukotrienes, lipids, biogenic amines) followed by chromatography, mass spectrometry, and isothermal titration calorimetry showed that DPTL binds with high affinity to carbocyclic TXA(2), TXA(2) mimetic (U-46619), TXB(2), PGH(2) mimetic (U-51605), PGD(2,) PGJ(2), and PGF(2α). It also interacts with 15(S)-HETE, being the first lipocalin described to date to bind to a derivative of 15-lipoxygenase. Binding was not observed to other prostaglandins (e.g. PGE(1), PGE(2), 8-iso-PGF(2α), prostacyclin), leukotrienes (e.g. LTB(4), LTC(4), LTD(4), LTE(4)), HETEs (e.g. 5(S)-HETE, 12(S)-HETE, 20-HETE), lipids (e.g. arachidonic acid, PAF), and biogenic amines (e.g. ADP, serotonin, epinephrine, norepinephrine, histamine). Consistent with its binding specificity, DPTL prevents contraction of rat uterus stimulated by PGF(2α) and induces relaxation of aorta previously contracted with U-46619. Moreover, it inhibits angiogenesis mediated by 15(S)-HETE and did not enhance inhibition of collagen-induced platelet aggregation by SQ29548 (TXA(2) antagonist) and indomethacin. A 3-D model for DPTL and pallidipin is presented that indicates the presence of a conserved Arg(39) and Gln(135) in the binding pocket of both lipocalins. Results suggest that DPTL blocks platelet aggregation, vasoconstriction, and angiogenesis through binding to distinct eicosanoids involved in inflammation.     

Inflammation, immunity, and vaccines for Helicobacter

Helicobacter pylori represents the major etiologic agent of gastritis, gastric, and duodenal ulcer disease and can cause gastric cancer and mucosa-associated lymphoid tissue B-cell lymphoma. It is clear that the consequences of infection reflect diverse outcomes of the interaction of bacteria and host immune system. The hope is that by deciphering the deterministic rules--if any--of this interplay, we will eventually be able to predict, treat, and ultimately prevent disease. Over the past year, research on the immunology of this infection started to probe the role of small noncoding RNAs, a novel class of immune response regulators. Furthermore, we learned new details on how infection is detected by innate pattern recognition receptors. Induction of effective cell-mediated immunity will be key for the development of a vaccine, and new work published analyzed the relevance and contribution of CD4 T helper cell subsets to the immune reaction. Th17 cells, which are also induced during natural infection, were shown to be particularly important for vaccination. Cost-efficiency of vaccination was re-assessed and confirmed. Thus, induction and shaping of the effector roles of such protective Th populations will be a target of the newly described vaccine antigens, formulations, and modes of application that we also review here.     

Canopy and litter ant assemblages share similar climate–species density relationships

Tropical forest canopies house most of the globe''s diversity, yet little is known about global patterns and drivers of canopy diversity. Here, we present models of ant species density, using climate, abundance and habitat (i.e. canopy versus litter) as predictors. Ant species density is positively associated with temperature and precipitation, and negatively (or non-significantly) associated with two metrics of seasonality, precipitation seasonality and temperature range. Ant species density was significantly higher in canopy samples, but this difference disappeared once abundance was considered. Thus, apparent differences in species density between canopy and litter samples are probably owing to differences in abundance–diversity relationships, and not differences in climate–diversity relationships. Thus, it appears that canopy and litter ant assemblages share a common abundance–diversity relationship influenced by similar but not identical climatic drivers.