Quantitative Analysis of Tau-Microtubule Interaction Using FRET

The interaction between the microtubule associated protein, tau and the microtubules is investigated. A fluorescence resonance energy transfer (FRET) assay was used to determine the distance separating tau to the microtubule wall, as well as the binding parameters of the interaction. By using microtubules stabilized with Flutax-2 as donor and tau labeled with rhodamine as acceptor, a donor-to-acceptor distance of 54 ± 1 Å was found. A molecular model is proposed in which Flutax-2 is directly accessible to tau-rhodamine molecules for energy transfer. By titration, we calculated the stoichiometric dissociation constant to be equal to 1.0 ± 0.5 µM. The influence of the C-terminal tails of αβ-tubulin on the tau-microtubule interaction is presented once a procedure to form homogeneous solution of cleaved tubulin has been determined. The results indicate that the C-terminal tails of α- and β-tubulin by electrostatic effects and of recruitment seem to be involved in the binding mechanism of tau.     

In vitro evaluation of S-(+)-ibuprofen as drug candidate for intra-articular drug delivery system

Intra-articular drug delivery systems (DDSs) are envisaged as interesting alternative to locally release non-steroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen to reduce pain in patients with osteoarthritis. The present study examines the efficacy of S-(+)-ibuprofen on cartilage degradation as drug candidate for DDS loading. Humeral cartilage and joint capsule explants were collected from healthy sheep shoulder joints and they were cultured in mono- or in co-culture for 13?days with LPS in combination with S-(+)-ibuprofen at 50?µM and 1?mM. S-(+)-ibuprofen (50?µM) blocked prostaglandins production in LPS-activated explants but did not reduce cartilage degradation. By contrast, 1?mM S-(+)-ibuprofen treatment of cartilage explants reduced nitric oxide synthesis by 51% (p?=?0.0072), proteoglycans degradation by 35% (p?=?0.0114) and expression of serum amyloid protein – the main protein induced upon LPS challenge – by 44% (p?相似文献   

Trans10,cis15 18:2 Isolated from Beef Fat Does Not Have the Same Anti-Adipogenic Properties as Trans10,cis12–18:2 in 3T3-L1 Adipocytes

During ruminal biohydrogenation of α‐linolenic acid, a non‐conjugated non‐methylene interrupted dienoic acid is formed containing a t10 double bond, namely t10,c15–18:2. The present study was designed to examine whether t10,c15–18:2 would exert similar anti‐adipogenic effects compared to t10,c12–18:2 in 3T3‐L1 adipocytes. Differentiated 3T3‐L1 adipocytes were treated with 35 or 70 µM of LNA, t10,c12–18:2, t10,c15–18:2, or bovine serum albumin (BSA) vehicle control for 120 h. Cellular triacylglycerol and protein were quantified using commercial colorimetric kits. Cells were analyzed for fatty acid composition and gene expression using gas chromatography and quantitative PCR, respectively. Trans10,cis12–18:2 decreased (P < 0.05) the adipocyte triacylglycerol (TAG) content, which was mainly related to a reduction in saturated fatty acids (SFA; e.g., 16:0 and 15:0) and cis monounsaturated fatty acids (c‐MUFA; e.g., c9–16:1 and c9–18:1). Trans10,cis12 also decreased (P < 0.05) the expression of genes related to fatty acid synthesis (ACACA, FASN), delta‐9 desaturation (SCD1), fatty acid elongation (ELOVL5), and fatty acid uptake (LPL) and upregulated (P < 0.05) the expression of the rate‐liming enzyme involved in fatty acid β‐oxidation (CPT1). In contrast, LNA and t10,c15–18:2 did not affect the gene expression and cellular content of the TAG, SFA, c‐MUFA, or SCD1 indices in adipocytes. Our findings suggest that t10,c15–18:2, despite having structural similarity to t10,c12–18:2 (presence of a trans‐10 double bond), does not exert anti‐adipogenic effects in 3T3‐L1 adipocytes.     

ASGPR‐Mediated Uptake of Multivalent Glycoconjugates for Drug Delivery in Hepatocytes

Liver cells are an essential target for drug delivery in many diseases. The hepatocytes express the asialoglycoprotein receptor (ASGPR), which promotes specific uptake by means of N‐acetylgalactosamine (GalNAc) recognition. In this work, we designed two different chemical architectures to treat Wilson's disease by intracellular copper chelation. Two glycoconjugates functionalized with three or four GalNAc units each were shown to enter hepatic cells and chelate copper. Here, we studied two series of compounds derived from these glycoconjugates to find key parameters for the targeting of human hepatocytes. Efficient cellular uptake was demonstrated by flow cytometry using HepG2 human heptic cells that express the human oligomeric ASGPR. Dissociation constants in the nanomolar range showed efficient multivalent interactions with the receptor. Both architectures were therefore concluded to be able to compete with endogeneous asialoglycoproteins and serve as good vehicles for drug delivery in hepatocytes.     

Vertically aligned carbon nanotube‐based composite: Elaboration and monitoring of the nanotubes alignment

We present the different elaboration steps of a composite formed of carbon nanotubes (CNT) carpet embedded in an epoxy polymer. Detailed characterization at each step of the elaboration process is performed. The good alignment of CNT in as‐grown carpets is kept all along the elaboration process of the composite, as it is measured at both macro and microscopic scales by X‐ray scattering. We also ensured by X‐ray fluorescence measurements that the iron‐based catalyst particles used for the synthesis were removed from the carpet after a high temperature post‐annealing treatment. These measurements give valuable information for further applications involving unidirectional nanotube composites and membranes, where CNT alignment is a key parameter. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 39730.     

A family of parallel QR factorization algorithms

Rapid computation of the QR factorization of a matrix is fundamental to many scientific and engineering problems. The paper presents a family of algorithms parameterized by the number of processors available P, arithmetic grain aggregation parameters g₁, g₂, …, g_P, and communication grain aggregation parameter h, which computer the QR factorization of a matrix A ∈ C^{m × n} with minimal latency. The approach is particularly well suited for dedicated distributed memory architectures such as linear arrays of INMOS Transputers, Texas Instruments C40s or Analog Devices 21060s.