Interdependent Regulation of Polycystin Expression Influences Starvation-Induced Autophagy and Cell Death

Autosomal dominant polycystic kidney disease (ADPKD) is mainly caused by deficiency of polycystin-1 (PC1) or polycystin-2 (PC2). Altered autophagy has recently been implicated in ADPKD progression, but its exact regulation by PC1 and PC2 remains unclear. We therefore investigated cell death and survival during nutritional stress in mouse inner medullary collecting duct cells (mIMCDs), either wild-type (WT) or lacking PC1 (PC1KO) or PC2 (PC2KO), and human urine-derived proximal tubular epithelial cells (PTEC) from early-stage ADPKD patients with PC1 mutations versus healthy individuals. Basal autophagy was enhanced in PC1-deficient cells. Similarly, following starvation, autophagy was enhanced and cell death reduced when PC1 was reduced. Autophagy inhibition reduced cell death resistance in PC1KO mIMCDs to the WT level, implying that PC1 promotes autophagic cell survival. Although PC2 expression was increased in PC1KO mIMCDs, PC2 knockdown did not result in reduced autophagy. PC2KO mIMCDs displayed lower basal autophagy, but more autophagy and less cell death following chronic starvation. This could be reversed by overexpression of PC1 in PC2KO. Together, these findings indicate that PC1 levels are partially coupled to PC2 expression, and determine the transition from renal cell survival to death, leading to enhanced survival of ADPKD cells during nutritional stress.     

A state space analysis of propose-and-revise

A central research topic in the area of knowledge engineering is the reuse of problem-solving methods for developing knowledge based systems. For being able to reuse a problem-solving method it is important to know under which restrictions a problem-solving method is appropriate to solve a given problem. This paper describes the problem-solving method propose-and-revise as well as the way this problem-solving method searches in its problem space for a solution. A quantitative analysis of the efficiency of this search process is given. Additionally, task and domain specific properties and restrictions and their influence on the efficiency of the search process are considered. For these purposes an instance of the problem-solving method is transformed to a corresponding instance of a Stanford Research Institute Problem Solver (STRIPS) planning system. Then the problem-solving method is considered as an additional control strategy for such a planning system. By this way the various insights and analysis results which are available in the area of planning systems may be exploited for the analysis of problem-solving methods. ©1999 John Wiley & Sons, Inc.     

How background noise shifts eigenvectors and increases eigenvalues in DTI

Indroduction  The signal-to-noise ratio of in vivo diffusion tensor imaging (DTI) is usually very limited, especially if high resolution data is acquired. In a variety of settings, the signal of diffusion weighted images can drop below the background noise level yielding an underestimated diffusion constant. In this work, we report two new artefacts in DTI that are important in this regime. Methods  Both artifacts are described analytically and numerically and are demonstrated in DTI phantoms and in subjects in vivo. Results  First, eigenvectors are systematically shifted towards distinct ‘attractive’ orientations of the gradient scheme. Second, certain eigenvalues can be overestimated due to the underestimation of the measured diffusion, which can result in the misordering of eigenvalues Discussion  We show that these effects are relevant for current clinical settings of DTI.     

Attentional retraining procedures: Manipulating early or late components of attentional bias?

According to cognitive models of anxiety disorders, attentional bias for threatening information is a vulnerability factor to the etiology and maintenance of anxiety. A recently developed methodology to reduce attentional bias has been found to reduce emotional reactivity and anxiety. The present study aimed at identifying the effects of this attentional bias reduction on early and later stages of threat processing. Undergraduates were allocated to an attentional bias reduction (n = 23) versus control condition (n = 25). It was found that attentional bias reduction influenced late but not early stages of threat processing. This finding is of theoretical importance in relation to studies on the causal role of attentional bias and emotional reactivity. Moreover, the present findings also bear relevance to the clinical application of attentional retraining procedures. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Pyrite Weathering in Reclaimed Shale Overburden at an Oil Sands Mine near Fort McMurray,Canada

Saline-sodic shale overburden associated with oil sand mining is a potential source of salt release to surface water and groundwater and can lead to salinization and/or sodification of reclamation covers. Weathering of shale overburden due to oxidation of sulphide minerals within the shale leads to sulphate (SO₄ ^2?) production and increased salinity. The controls on the rates of weathering of a shale overburden dump in the oil sands region of northern Alberta were determined from soil chemistry sampling and in situ monitoring of pore gases (O₂, CO₂, CH₄) in three shallow profiles (1.9–4.45 m deep) and one deep (25 m deep) profile under reclamation covers of varying thickness. Oxidation, defined by the depth over which O₂ concentrations were depleted, reached depths of approximately 1.1 m under the reclamation soil covers over a 6 year period after dump placement. Calculations of SO₄ ^2? production rates and weathering depths were consistent with numerical simulations of the diffusion and subsequent consumption of atmospheric O₂ in the overburden. The rate of SO₄ ^2? production during the 6 year weathering period estimated from direct measurements of solids chemistry ranged from 0.70 to 8.3 g m^?2 day^?1. The rates calculated from the oxygen diffusion models were within that same range, between 1.6 and 4.1 g m^?2 day^?1.     

Real-time assessment of fluid flow generated by appendage movements of Daphnia using standing square-wave chronamperometry

Standing square-wave chronoamperometry (SSWCA) was applied to the analysis of the microfluid flow generated by the movement of the appendages of the Crustacea Daphnia. This novel approach provided for the first time real-time assessment and analysis of the breathing rate/fluid flow of individual organisms. An electrochemical tracer was delivered into the fluid inflow of the organism and a carbon fiber microelectrode placed in the fluid outflow's path. The variation of the net concentration/flux of the electroactive tracer, dopamine, at the electrode surface was measured with SSWCA. The observed chronoamperometric peaks (with fine structure) of the outflow are seen as a direct representation of appendage movement and, too, the workings and responses of the organism to its environment, e.g., external stimuli such as food or chemicals. It was concluded that SSWCA follows primarily the variation of the convective component of the Nernst-Plank equation for flux and, to lesser extent, diffusion and migration. In this work, SSWCA can clearly be used to monitor changes in the Daphnia-generated fluid outflow on a different time scale than was previously possible. This new application of SSWCA is faster and likely more accurate than using high-speed video.     

Indium tin oxide-coated glass modified with reduced graphene oxide sheets and gold nanoparticles as disposable working electrodes for dopamine sensing in meat samples

Sensitive, rapid, and accurate detection of dopamine (DA) at low cost is needed for clinical diagnostic and therapeutic purposes as well as to prevent illegal use of DA in animal feed. We employed a simple approach to synthesize reduced graphene oxide sheets (rGOS) and gold nanoparticles (AuNPs) at room temperature on indium tin oxide-coated glass (ITO) slides as disposable working electrodes for sensing DA. Graphene oxide (GO) was directly reduced on ITO to remove oxygenated species via a rapid and green process without using chemical reducing reagents. AuNPs were electrochemically deposited in situ on rGOS-ITO with fairly uniform density and size. The sensitivity of the AuNPs-rGOS-ITO sensor for DA detection is 62.7 μA mM(-1) cm(-2) with good selectivity against common electrochemically interfering species such as ascorbic acid (AA) and uric acid (UA), and the detection limit measured by differential pulse voltammetry (DPV), at a signal-noise ratio of 3, was 6.0 × 10(-8) M. The electrochemical catalysis of DA was proven to be a surface process with an electron transfer coefficient (α) of 0.478 and a rate constant (k(s)) of 1.456 s(-1). It correlates well with the conventional UV-vis spectrophotometric approach (R = 0.9973) but with more than thrice the dynamic range (up to 4.5 mM). The sensor also exhibited good stability and capability to detect DA in beef samples, and thus is a promising candidate for simple and inexpensive sub-nanomolar detection of DA, especially in the presence of UV-absorbing compounds.     

Strain-level characterization of nonstarter lactic acid bacteria in Norvegia cheese by high-resolution melt analysis

The nonstarter lactic acid bacteria (NSLAB) constitute an important microbial group found during cheese ripening and they are thought to be fundamental to the quality of cheese. Rapid and accurate diagnostic tests for NSLAB are important for cheese quality control and in understanding the cheese ripening process. Here, we present a novel rapid approach for strain-level characterization through combined 16S rRNA gene and repetitive sequence-based high-resolution melt analysis (HRM). The approach was demonstrated through the characterization of 94 isolates from Norvegia, a Gouda-type cheese. The HRM profiles of the V1 and V3 variable regions of the 16S rRNA gene of the isolates were compared with the HRM profiles of 13 reference strains. The HRM profile comparison of the V1 and V3 regions of the 16S rRNA gene allowed discrimination of isolates and reference strains. Among the cheese isolates, Lactobacillus casei/paracasei (62 isolates) and Lactobacillus plantarum/Lactobacillus pentosus (27 isolates) were the dominant species, whereas Lactobacillus curvatus/Lactobacillus sakei were found occasionally (5 isolates). The HRM profiling of repetitive sequence-based PCR using the (GTG)(5) primer was developed for strain-level characterization. The clustering analysis of the HRM profiles showed high discriminatory power, similar to that of cluster analysis based on the gel method. In conclusion, the HRM approach in this study may be applied as a fast, accurate, and reproducible method for characterization of the NSLAB microflora in cheese and may be applicable to other microbial environments following selective plate culturing.     

The effect of crowding stress on bacterial growth and sensory properties of chilled Atlantic salmon fillets

Abstract: Atlantic salmon were subjected to minimal preslaughter crowding stress (Control), short‐term crowding for 20 min (SS‐group), or long‐term crowding for 24 h (LS‐group). The fish were filleted prerigor, cut into 270 g pieces, and packaged in modified atmosphere (60% CO₂ and 40% N₂). Fillet quality analyses were determined during 22 d of storage at 0.3 °C. Bacterial growth and unpleasant sensory properties increased earlier in the LS‐group. The negative effects of long‐term preslaughter stress were more pronounced for raw than cooked samples, and more pronounced for odor than flavor. Sequence analyses of bacterial DNA at the end of storage revealed that 100% of the bacteria were comprised by Photobacterium phosphoreum of the SS‐ and LS‐group, whereas the Control group also contained 21% of Carnobacterium maltaromaticum (lactic acid bacteria, LAB). Counting of LAB, using Man‐Rogosa‐Sharke agar, similarly showed higher numbers of the Control group after 15 d of storage. A total bacterial count of log 6 CFU/g was observed after 15 d of storage of the LS‐group, which was 3 and 7 d earlier compared with the Control and SS‐group, respectively. Fillet color, texture, and liquid losses were not negatively affected by preslaughter crowding stress. From the sensory and bacterial analyses, it is concluded that long‐term crowding stress accelerates bacterial growth and development of unpleasant sensory properties, hence reduces the shelf life of prerigor modified atmosphere packaged (MAP) salmon. Practical Application: Stressful handling of Atlantic salmon before slaughter resulted in faster reduction of fresh taste and smell, faster bacterial growth, and hence shorter shelf life. The deteriorative effects were more pronounced of raw compared to cooked salmon. Therefore, salmon should be handled carefully in connection with slaughter to avoid impaired welfare and fillet quality, in particularly for fish that is consumed raw, such as sushi.