The O-glycomap of lubricin, a novel mucin responsible for joint lubrication, identified by site-specific glycopeptide analysis

The lubricative, heavily glycosylated mucin-like synovial glycoprotein lubricin has previously been observed to contain glycosylation changes related to rheumatoid and osteoarthritis. Thus, a site-specific investigation of the glycosylation of lubricin was undertaken, in order to further understand the pathological mechanisms involved in these diseases. Lubricin contains an serine/threonine/proline (STP)-rich domain composed of imperfect tandem repeats (EPAPTTPK), the target for O-glycosylation. In this study, using a liquid chromatography–tandem mass spectrometry approach, employing both collision-induced and electron-transfer dissociation fragmentation methods, we identified 185 O-glycopeptides within the STP-rich domain of human synovial lubricin. This showed that adjacent threonine residues within the central STP-rich region could be simultaneously and/or individually glycosylated. In addition to core 1 structures responsible for biolubrication, core 2 O-glycopeptides were also identified, indicating that lubricin glycosylation may have other roles. Investigation of the expression of polypeptide N-acetylgalactosaminyltransferase genes was carried out using cultured primary fibroblast-like synoviocytes, a cell type that expresses lubricin in vivo. This analysis showed high mRNA expression levels of the less understood polypeptide N-acetylgalactosaminyltransferase 15 and 5 in addition to the ubiquitously expressed polypeptide N-acetylgalactosaminyltransferase 1 and 2 genes. This suggests that there is a unique combination of transferase genes important for the O-glycosylation of lubricin. The site-specific glycopeptide analysis covered 82% of the protein sequence and showed that lubricin glycosylation displays both micro- and macroheterogeneity. The density of glycosylation was shown to be high: 168 sites of O-glycosylation, predominately sialylated, were identified. These glycosylation sites were focused in the central STP-rich region, giving the domain a negative charge. The more positively charged lysine and arginine residues in the N and C termini suggest that synovial lubricin exists as an amphoteric molecule. The identification of these unique properties of lubricin may provide insight into the important low-friction lubricating functions of lubricin during natural joint movement

Pre-Prandial Vinegar Ingestion Improves Two-Hour Glucose Control in Older, Type II Diabetics More Than Post-Prandial Walking

Background: Exercise engagement benefits diabetic patients through an insulin-like effect on muscle. Literature indicates that vinegar consumption may lower blood glucose levels. It is not currently clarified whether a relative amount of vinegar ingestion or a walking bout is more effective at controlling glucose in older, Type II diabetics during the acute phase following a meal. Purpose: The aim was to directly compare the impact of preferred-pace walking (15 min, postprandial) versus ingestion of a relative quantity of vinegar (0.3 g/kg) on two-hour glucose control. Methods: The two arms of the trial were completed in a randomized, crossover manner. Six Type II diabetic patients (Females = 5; Males = 1; Age = 70.5 ± 9.0 yrs.) enrolled and underwent baseline finger pricks to establish glucose levels. The test meal consisted of an 85 g bagel, 13 g of butter, and 237 mL of orange juice. On the respective days, the vinegar was diluted into 59 mL of orange juice and ingested before the meal or the subject completed a 15-min walk at 15 min post-meal. For both trials, glucose was checked every 30 min following the test meal. Results: One subject was removed from all present analysis due to medication-related non-compliance. For the vinegar trial, the resting heart rate was 72.0 (± 9.5) and baseline, 30-, 60-, 90-, and 120-min average blood glucose levels were: 117 (±12), 149 (±39), 172 (±49), 185 (±49), and 180 (±44) mg/dl. For the preferred walking speed phase, the resting heart rate was 75.5 (±15.6) and baseline, 30-, 60-, 90-, and 120-min average glucose levels were: 113 (±10), 147 (±53), 180 (±53), 208 (±72), and 206 (±71) mg/dl. Preferred walking speed was found to average 3.1 (± 1.5) kph and total steps averaged 1418 (±376). The between-arm comparison of glucose at 120-min trended towards significance (p = 0.081). Conclusions: Compared with a bout of walking, a relative quantity of vinegar may serve as a more suitable mechanism for older Type II diabetics to control acute spikes in glucose after a high carbohydrate meal. With an adequately-powered analysis, between-arm comparisons at multiple time-points would likely have achieved statistical significance. Nevertheless, the meaningfulness of the glucose control exhibited should not be lost due to the lack of statistical significance. Finally, the slow absolute preferred walking pace of many older adults may undermine the ability for walking to result in sufficient energy expenditure capable of subsequent glucose control

Decrease of core 2 O-glycans on synovial lubricin in osteoarthritis reduces galectin-3 mediated crosslinking

The synovial fluid glycoprotein lubricin (also known as proteoglycan 4) is a mucin-type O-linked glycosylated biological lubricant implicated to be involved in osteoarthritis (OA) development. Lubricin\u27s ability to reduce friction is related to its glycosylation consisting of sialylated and unsialylated Tn-antigens and core 1 and core 2 structures. The glycans on lubricin have also been suggested to be involved in crosslinking and stabilization of the lubricating superficial layer of cartilage by mediating interaction between lubricin and galectin-3. However, with the spectrum of glycans being found on lubricin, the glycan candidates involved in this interaction were unknown. Here, we confirm that the core 2 O-linked glycans mediate this lubricin-galectin-3 interaction, shown by surface plasmon resonance data indicating that recombinant lubricin (rhPRG4) devoid of core 2 structures did not bind to recombinant galectin-3. Conversely, transfection of Chinese hamster ovary cells with the core 2 GlcNAc transferase acting on a mucin-type O-glycoprotein displayed increased galectin-3 binding. Both the level of galectin-3 and the galectin-3 interactions with synovial lubricin were found to be decreased in late-stage OA patients, coinciding with an increase in unsialylated core 1 O-glycans (T-antigens) and Tn-antigens. These data suggest a defect in crosslinking of surface-active molecules in OA and provide novel insights into OA molecular pathology

River patterns reveal two stages of landscape evolution at an oblique convergent margin, Marlborough Fault System, New Zealand

Here we examine the landscape of New Zealand's Marlborough Fault System (MFS), where the Australian and Pacific plates obliquely collide, in order to study landscape evolution and the controls on fluvial patterns at a long-lived plate boundary. We present maps of drainage anomalies and channel steepness, as well as an analysis of the plan-view orientations of rivers and faults, and we find abundant evidence of structurally controlled drainage that we relate to a history of drainage capture and rearrangement in response to mountain-building and strike-slip faulting. Despite clear evidence of recent rearrangement of the western MFS drainage network, rivers in this region still flow parallel to older faults, rather than along orthogonal traces of younger, active strike-slip faults. Such drainage patterns emphasize the importance of river entrenchment, showing that once rivers establish themselves along a structural grain, their capture or avulsion becomes difficult, even when exposed to new weakening and tectonic strain. Continued flow along older faults may also indicate that the younger faults have not yet generated a fault damage zone with the material weakening needed to focus erosion and reorient rivers. Channel steepness is highest in the eastern MFS, in a zone centered on the Kaikōura ranges, including within the low-elevation valleys of main stem rivers and at tributaries near the coast. This pattern is consistent with an increase in rock uplift rate toward a subduction front that is locked on its southern end. Based on these results and a wealth of previous geologic studies, we propose two broad stages of landscape evolution over the last 25 million years of orogenesis. In the eastern MFS, Miocene folding above blind thrust faults generated prominent mountain peaks and formed major transverse rivers early in the plate collision history. A transition to Pliocene dextral strike-slip faulting and widespread uplift led to cycles of river channel offset, deflection and capture of tributaries draining across active faults, and headward erosion and captures by major transverse rivers within the western MFS. We predict a similar landscape will evolve south of the Hope Fault, as the locus of plate boundary deformation migrates southward into this region with time.</p

Benfotiamine and Cognitive Decline in Alzheimer’s Disease: Results of a Randomized Placebo-Controlled Phase IIa Clinical Trial

Background: In preclinical models, benfotiamine efficiently ameliorates the clinical and biological pathologies that define Alzheimer’s disease (AD) including impaired cognition, amyloid-β plaques, neurofibrillary tangles, diminished glucose metabolism, oxidative stress, increased advanced glycation end products (AGE), and inflammation. Objective:To collect preliminary data on feasibility, safety, and efficacy in individuals with amnestic mild cognitive impairment (aMCI) or mild dementia due to AD in a placebo-controlled trial of benfotiamine. Methods:A twelve-month treatment with benfotiamine tested whether clinical decline would be delayed in the benfotiamine group compared to the placebo group. The primary clinical outcome was the Alzheimer’s Disease Assessment Scale-Cognitive Subscale (ADAS-Cog). Secondary outcomes were the clinical dementia rating (CDR) score and fluorodeoxyglucose (FDG) uptake, measured with brain positron emission tomography (PET). Blood AGE were examined as an exploratory outcome. Results:Participants were treated with benfotiamine (34) or placebo (36). Benfotiamine treatment was safe. The increase in ADAS-Cog was 43% lower in the benfotiamine group than in the placebo group, indicating less cognitive decline, and this effect was nearly statistically significant (p = 0.125). Worsening in CDR was 77% lower (p = 0.034) in the benfotiamine group compared to the placebo group, and this effect was stronger in the APOE ɛ4 non-carriers. Benfotiamine significantly reduced increases in AGE (p = 0.044), and this effect was stronger in the APOE ɛ4 non-carriers. Exploratory analysis derivation of an FDG PET pattern score showed a treatment effect at one year (p = 0.002). Conclusion:Oral benfotiamine is safe and potentially efficacious in improving cognitive outcomes among persons with MCI and mild AD

Measurement of the prompt J/psi and psi(2S) polarizations in pp collisions at sqrt(s) = 7 TeV

The polarizations of prompt J/psi and psi(2S) mesons are measured in proton-proton collisions at sqrt(s) = 7 TeV, using a dimuon data sample collected by the CMS experiment at the LHC, corresponding to an integrated luminosity of 4.9 inverse femtobarns. The prompt J/psi and psi(2S) polarization parameters lambda[theta], lambda[phi], and lambda[theta, phi], as well as the frame-invariant quantity lambda(tilde), are measured from the dimuon decay angular distributions in three different polarization frames. The J/psi results are obtained in the transverse momentum range 14 &lt; pt &lt; 70 GeV, in the rapidity intervals abs(y) &lt; 0.6 and 0.6 &lt; abs(y) &lt; 1.2. The corresponding psi(2S) results cover 14 &lt; pt &lt; 50 GeV and include a third rapidity bin, 1.2 &lt; abs(y) &lt; 1.5. No evidence of large transverse or longitudinal polarizations is seen in these kinematic regions, which extend much beyond those previously explored

The CMS Phase-1 pixel detector upgrade

The CMS detector at the CERN LHC features a silicon pixel detector as its innermost subdetector. The original CMS pixel detector has been replaced with an upgraded pixel system (CMS Phase-1 pixel detector) in the extended year-end technical stop of the LHC in 2016/2017. The upgraded CMS pixel detector is designed to cope with the higher instantaneous luminosities that have been achieved by the LHC after the upgrades to the accelerator during the first long shutdown in 2013–2014. Compared to the original pixel detector, the upgraded detector has a better tracking performance and lower mass with four barrel layers and three endcap disks on each side to provide hit coverage up to an absolute value of pseudorapidity of 2.5. This paper describes the design and construction of the CMS Phase-1 pixel detector as well as its performance from commissioning to early operation in collision data-taking.Peer reviewe

Alignment of the CMS tracker with LHC and cosmic ray data

© CERN 2014 for the benefit of the CMS collaboration, published under the terms of the Creative Commons Attribution 3.0 License by IOP Publishing Ltd and Sissa Medialab srl. Any further distribution of this work must maintain attribution to the author(s) and the published article's title, journal citation and DOI.The central component of the CMS detector is the largest silicon tracker ever built. The precise alignment of this complex device is a formidable challenge, and only achievable with a significant extension of the technologies routinely used for tracking detectors in the past. This article describes the full-scale alignment procedure as it is used during LHC operations. Among the specific features of the method are the simultaneous determination of up to 200 000 alignment parameters with tracks, the measurement of individual sensor curvature parameters, the control of systematic misalignment effects, and the implementation of the whole procedure in a multi-processor environment for high execution speed. Overall, the achieved statistical accuracy on the module alignment is found to be significantly better than 10μm