The pleckstrin homology domain of oxysterol-binding protein recognises a determinant specific to Golgi membranes

AbstractBackground: Peripheral membrane proteins are targeted to the cytoplasmic face of specific intracellular membranes. The organelle-specific ligands recognised by peripheral proteins include both other proteins and lipids. Oxysterol-binding protein (OSBP) translocates from the cytoplasm to the Golgi apparatus on binding oxygenated derivatives of cholesterol. The mechanism by which OSBP recognises the Golgi is unknown. It does, however, contain a pleckstrin homology (PH) domain, which in other proteins has been found to mediate regulated membrane binding, although in all previously studied examples the binding is to the plasma membrane.Results: The PH domains of OSBP and of a yeast homologue, Osh1p, were sufficient to target fusion proteins specifically to mammalian Golgi membranes. In addition, high level expression disrupted Golgi architecture and prevented forward traffic of cargo protein. In vitro, the OSBP PH domain bound to Golgi membranes in a manner apparently dependent on phosphatidylinositol-4,5-bisphosphate (Pl(4,5)P2) or a related phosphatidylinositide. The OSBP PH domain bound to Pl(4,5)P2 in liposomes with a submicromolar dissociation constant.Conclusions: The PH domains of OSBP and its yeast homologue recognise a determinant which is specific to Golgi membranes and important for Golgi function. The determinant appears to be a combination of a phosphatidylinositol polyphosphate and a second, Golgi-specific feature

Comparison of Inter and Intra-Operator Differences for Cephalometric Landmark Identification on Three-Dimensional CBCT Images using Pro Plan CMF

Objective: To establish reliability of cephalometric landmark identification in threedimensions using ProPlan CMF software. Methods: Two orthodontist identified a series of 33 cephalometric landmarks on 20 CBCT scans of Class I, pre-orthodontic patients and repeated the landmark identification about two months later. Intraclass correlations (ICC) were calculated by landmark in the X, Y, and Z dimensions and F-test were used to assess difference in landmark location in the X, Y, and Z dimensions. Results: The majority of landmarks had good to excellent ICC for both inter- and intraobserver reliability. F-test also showed the majority of landmarks had no significant difference between the observers. Conclusion: Most landmarks showed good to very good reliability and reproducibility using ProPlan CMF, with some landmarks proving more reliable than others and further research is needed to establish the utility and practicality of three-dimensional cephalometrics as a common diagnostic tool in orthodontics

A Double-Blind, Placebo-Controlled Withdrawal Trial of Dexmethylphenidate Hydrochloride in Children with Attention Deficit Hyperactivity Disorder

Objectives: d,l-threo-methylphenidate HCl (d,l-MPH) is the most common treatment of attention deficit hyperactivity disorder (ADHD). A previous report showed placebo-controlled efficacy for the purified disomer (dexmethylphenidate hydrochloride, d-MPH, Focalin™) with a 2:1 potency compared to dl, and suggested a 6-hour duration of action. This study complements that report by studying the effect of placebo-controlled discontinuation and retesting the duration of action. Methods: A 6-week, open-label titration of d-MPH (2.5–10 mg twice-a-day) was followed by a doubleblind, placebo-controlled, 2-week withdrawal study of responders. Results: In the open titration, 82% of the 89 enrolled patients achieved a Clinical Global Impression— Improvement (CGI-I) rating of much or very much improved. Only 5 patients discontinued for adverse events. Seventy-five patients continued into the placebo-controlled discontinuation. For the randomly assigned d-MPH (n = 35) and placebo (n = 40) groups, mean ages, respectively, were 10.1 ± 2.9 and 9.9 ± 2.7 years, 86% and 78% were male, and 70.6% and 80.0% took the ceiling dose of 10 mg twice-daily, respectively. Each group had 80% combined-type ADHD and 20% inattentive type. By the end of the 2- week, placebo-masked withdrawal, significantly more placebo patients (24 of 39) than d-MPH continuers (6 of 35) relapsed (61.5% versus 17.1%, p = 0.001). Compared to d-MPH continuers, placebo patients deteriorated significantly more in the 2-week period on teacher ratings of the 18 ADHD symptoms rated 0– 3 (p = 0.028), the 3 p.m. and 6 p.m. parent ADHD symptom ratings (p = 0.0026 and p = 0.0381, respectively), and clinic (2–3 p.m.) and home (6 p.m.) Math Tests (p = 0.024 and p < 0.0001, respectively). The 6 p.m. scores replicated the significant effect at 6 hours reported in the previous study. Conclusions: d-MPH is safe, tolerable, and effective, with a 6-hour duration of effect suggested by the significant difference from placebo at 6 hours on a double-blind discontinuation

Mooring observations from the Oregon continental shelf : April-September 1999 : a component of the prediction of wind-driven coastal circulation project

This report documents oceanographic and meteorological measurements made from instruments deployed on four moorings over the continental shelf west of Oregon, from spring through summer, 1999. These moorings were a component of an observational and numerical modeling program to study the response of the coastal ocean to wind forcing. The Dynamics and Prediction of Wind-Driven Coastal Circulation was funded by the National Oceanographic Partnership Program (NOPP) with the principal goal to develop nowcast and forecast systems for wind-driven coastal flow fields. The observational program was designed to provide measurements that would allow testing and improvement of the modeling capability. See http://www.oce.orst.edu/po/research/nopp/, http://diana.OCE.ORST.EDU/cmoweb/nopp/, and Austin et al. (2000) for description of the modeling program and a description of other aspects of the observational program. This report is divided into two sections. The first section contains descriptions of the instrumentation deployed on the NOPP moorings including locations, sampling rates, and calibrations. The second section contains plots of the observations. Several views of the time series recorded by the moorings are presented. Time series of vertically separated velocity, temperature, and salinity are shown for each mooring. Velocity and temperature observations from the same depth on horizontally separated moorings are also shown. These data are presented as both 40-hour low-pass filtered and 1-hour low-pass filtered time series. Time is given as day of year 1999 in all of the time series plots; conversion to calendar date is provided in Table 5

CTGF drives autophagy, glycolysis and senescence in cancer-associated fibroblasts via HIF1 activation, metabolically promoting tumor growth

Previous studies have demonstrated that loss of caveolin-1 (Cav-1) in stromal cells drives the activation of the TGF-β signaling, with increased transcription of TGF-β target genes, such as connective tissue growth factor (CTGF). In addition, loss of stromal Cav-1 results in the metabolic reprogramming of cancer-associated fibroblasts, with the induction of autophagy and glycolysis. However, it remains unknown if activation of the TGF-β / CTGF pathway regulates the metabolism of cancer-associated fibroblasts. Therefore, we investigated whether CTGF modulates metabolism in the tumor microenvironment. For this purpose, CTGF was overexpressed in normal human fibroblasts or MDA-MB-231 breast cancer cells. Overexpression of CTGF induces HIF-1α-dependent metabolic alterations, with the induction of autophagy/mitophagy, senescence, and glycolysis. Here, we show that CTGF exerts compartment-specific effects on tumorigenesis, depending on the cell-type. In a xenograft model, CTGF overexpressing fibroblasts promote the growth of co-injected MDA-MB-231 cells, without any increases in angiogenesis. Conversely, CTGF overexpression in MDA-MB-231 cells dramatically inhibits tumor growth in mice. Intriguingly, increased extracellular matrix deposition was seen in tumors with either fibroblast or MDA-MB-231 overexpression of CTGF. Thus, the effects of CTGF expression on tumor formation are independent of its extracellular matrix function, but rather depend on its ability to activate catabolic metabolism. As such, CTGF-mediated induction of autophagy in fibroblasts supports tumor growth via the generation of recycled nutrients, whereas CTGF-mediated autophagy in breast cancer cells suppresses tumor growth, via tumor cell self-digestion. Our studies shed new light on the compartment-specific role of CTGF in mammary tumorigenesis, and provide novel insights into the mechanism(s) generating a lethal tumor microenvironment in patients lacking stromal Cav-1. As loss of Cav-1 is a stromal marker of poor clinical outcome in women with primary breast cancer, dissecting the downstream signaling effects of Cav-1 are important for understanding disease pathogenesis, and identifying novel therapeutic targets

Progression of mitral regurgitation in rheumatic valve disease : role of left atrial remodeling

Introduction: Mitral regurgitation (MR) is the most common valve abnormality in rheumatic heart disease (RHD) often associated with stenosis. Although the mechanism by which MR develops in RHD is primary, longstanding volume overload with left atrial (LA) remodeling may trigger the development of secondary MR, which can impact on the overall progression of MR. This study is aimed to assess the incidence and predictors of MR progression in patients with RHD. Methods: Consecutive RHD patients with non-severe MR associated with any degree of mitral stenosis were selected. The primary endpoint was a progression of MR, which was defined as an increase of one grade in MR severity from baseline to the last follow-up echocardiogram. The risk of MR progression was estimated accounting for competing risks. Results: The study included 539 patients, age of 46.2 ± 12 years and 83% were women. At a mean follow-up time of 4.2 years (interquartile range [IQR]: 1.2–6.9 years), 54 patients (10%) displayed MR progression with an overall incidence of 2.4 per 100 patient-years. Predictors of MR progression by the Cox model were age (adjusted hazard ratio [HR] 1.541, 95% CI 1.222–1.944), and LA volume (HR 1.137, 95% CI 1.054–1.226). By considering competing risk analysis, the direction of the association was similar for the rate (Cox model) and incidence (Fine-Gray model) of MR progression. In the model with LA volume, atrial fibrillation (AF) was no longer a predictor of MR progression. In the subgroup of patients in sinus rhythm, 59 had an onset of AF during follow-up, which was associated with progression of MR (HR 2.682; 95% CI 1.133–6.350). Conclusions: In RHD patients with a full spectrum of MR severity, progression of MR occurs over time is predicted by age and LA volume. LA enlargement may play a role in the link between primary MR and secondary MR in patients with RHD

The Milky Way's circular velocity curve between 4 and 14 kpc from APOGEE data

We measure the Milky Way's rotation curve over the Galactocentric range 4 kpc <~ R <~ 14 kpc from the first year of data from the Apache Point Observatory Galactic Evolution Experiment (APOGEE). We model the line-of-sight velocities of 3,365 stars in fourteen fields with b = 0 deg between 30 deg < l < 210 deg out to distances of 10 kpc using an axisymmetric kinematical model that includes a correction for the asymmetric drift of the warm tracer population (\sigma_R ~ 35 km/s). We determine the local value of the circular velocity to be V_c(R_0) = 218 +/- 6 km/s and find that the rotation curve is approximately flat with a local derivative between -3.0 km/s/kpc and 0.4 km/s/kpc. We also measure the Sun's position and velocity in the Galactocentric rest frame, finding the distance to the Galactic center to be 8 kpc < R_0 < 9 kpc, radial velocity V_{R,sun} = -10 +/- 1 km/s, and rotational velocity V_{\phi,sun} = 242^{+10}_{-3} km/s, in good agreement with local measurements of the Sun's radial velocity and with the observed proper motion of Sgr A*. We investigate various systematic uncertainties and find that these are limited to offsets at the percent level, ~2 km/s in V_c. Marginalizing over all the systematics that we consider, we find that V_c(R_0) 99% confidence. We find an offset between the Sun's rotational velocity and the local circular velocity of 26 +/- 3 km/s, which is larger than the locally-measured solar motion of 12 km/s. This larger offset reconciles our value for V_c with recent claims that V_c >~ 240 km/s. Combining our results with other data, we find that the Milky Way's dark-halo mass within the virial radius is ~8x10^{11} M_sun.Comment: submitted to Ap

Distant effect of assimilation of moored currents into a model of coastal wind-driven circulation off Oregon

An optimal interpolation (OI) sequential algorithm is implemented for a three-dimensional primitive equation model to assimilate current measurements from acoustic Doppler profilers moored on the Oregon shelf as a part of the Coastal Ocean Advances in Shelf Transport (COAST) upwelling experiment (May–August 2001). A stationary estimate of the forecast error covariance required by the OI is computed based on the error covariance in the model solution not constrained by data assimilation. Lagged model error covariances are used to account for the effect of previously assimilated data. The forecast error covariance has a shorter alongshore spatial scale than the model error covariance unconstrained by the data, as an effect of propagating dynamical modes. Assimilation of currents from one or two of the moorings located on the path of the upwelling jet helps to improve the model data rms error and correlation at the mooring sites located at an alongshore distance of 90 km, south or north from the assimilation sites. The coastal jet is deflected offshore over Heceta Bank, and assimilation of data from an inner-shelf mooring in the jet separation zone does not help to improve prediction in the far field. Larger improvements are obtained for the first part of the study period (yeardays 146–190). In the second part (days 191–237) the geometry of our limited area model possibly limits prediction accuracy. In numerical experiments involving assimilation of data from only one mooring the actual and expected rms error improvements are compared, providing a consistency test for the forecast error covariance.Keywords: upwelling, coastal ocean prediction, data assimilationKeywords: upwelling, coastal ocean prediction, data assimilatio