Interfering with the inhibitory mechanism of serpins: crystal structure of a complex formed between cleaved plasminogen activator inhibitor type 1 and a reactive-centre loop peptide

BACKGROUND: Plasminogen activator inhibitor type 1 (PAI-1) is an important endogenous regulator of the fibrinolytic system. Reduction of PAI-1 activity has been shown to enhance dissolution of blood clots. Like other serpins, PAI-1 binds covalently to a target serine protease, thereby irreversibly inactivating the enzyme. During this process the exposed reactive-centre loop of PAI-1 is believed to undergo a conformational change becoming inserted into beta sheet A of the serpin. Incubation with peptides from the reactive-centre loop transform serpins into a substrate for their target protease. It has been hypothesised that these peptides bind to beta sheet A, thereby hindering the conformational rearrangement leading to loop insertion and formation of the stable serpin-protease complex. RESULTS: We report here the 1.95 A X-ray crystal structure of a complex of a glycosylated mutant of PAI-1, PAI-1-ala335Glu, with two molecules of the inhibitory reactive-centre loop peptide N-Ac-TVASS-NH2. Both bound peptide molecules are located between beta strands 3A and 5A of the serpin. The binding kinetics of the peptide inhibitor to immobilised PAI-1-Ala335Glu, as monitored by surface plasmon resonance, is consistent with there being two different binding sites. CONCLUSIONS: This is the first reported crystal structure of a complex formed between a serpin and a serpin inhibitor. The localisation of the inhibitory peptide in the complex strongly supports the theory that molecules binding in the space between beta strands 3A and 5A of a serpin are able to prevent insertion of the reactive-centre loop into beta sheet A, thereby abolishing the ability of the serpin to irreversibly inactivate its target enzyme. The characterisation of the two binding sites for the peptide inhibitor provides a solid foundation for computer-aided design of novel, low molecular weight PAI-1 inhibitors.     

Morphology and mechanical properties of poly(propylene)/polystyrene blends compatibilized with polystyrene-block-poly(ethylene-co-propylene)

Compatibilizing effects of diblock copolymer polystyrene-block-poly(ethylene-co-propylene) (SEP) on the morphology and mechanical properties of immiscible blends of poly(propylene) (PP) and polystyrene (PS) were investigated. Notched impact strength, yield stress, elongation at yield and Young's modulus were determined as a function of different weight ratios of PP and PS and different amounts of added SEP as well. Scanning electron microscopy revealed a two-phase morphology of PP/PS blends, which exhibit poor mechanical properties. Even 2,5 wt.-% of SEP added to PP/PS blends can improve the notched impact strength and elongation at yield compared to non-compatibilized PP/PS blends. 10 wt.-% of SEP compatibilizer converted the brittle PP/PS blend to quite impactresistant polymeric material. Mechanical properties were improved because of the morphological changes and increased interfacial adhesion as a result of SEP localization between PP and PS phases. An analysis of yield stress data in terms of theoretical models showed that yield stress values of binary PP/PS blends can be predicted with Nielsen's model.     

Grinding induced effects in plasma sprayed zirconia coatings

The effect of grinding on the surface layer properties of ceria and yttria partially stabilized zirconia plasma-sprayed coatings (CePSZ, YPSZ, respectively) has been studied by X-ray diffraction methods. For this purpose, the modified model of line broadening analysis has been derived. The model considers elastic anisotropic properties along with more random paracrystal imperfections, both affecting X-ray line broadening. Grinding-induced microstructural changes were also studied using an estimation from the quantitative Orientation Distribution Function (ODF) texture. It was concluded, based on this work, that CePSZ ceramic is less mechanically stable compared to YPSZ. Consequently, more beneficial mechanical properties of a ground surface layer can be expected for CePSZ plasma-sprayed coatings.     

Diagnosing Czech Patients with Inherited Platelet Disorders

A single-center study was conducted on 120 patients with inherited disorders of primary hemostasis followed at our hematological center. These patients presented a variety of bleeding symptoms; however, they had no definitive diagnosis. Establishing a diagnosis has consequences for the investigation of probands in families and for treatment management; therefore, we aimed to improve the diagnosis rate in these patients by implementing advanced diagnostic methods. According to the accepted international guidelines at the time of study, we investigated platelet morphology, platelet function assay, light-transmission aggregometry, and flow cytometry. Using only these methods, we were unable to make a definitive diagnosis for most of our patients. However, next-generation sequencing (NGS), which was applied in 31 patients, allowed us to establish definitive diagnoses in six cases (variants in ANKRD26, ITGA2B, and F8) and helped us to identify suspected variants (NBEAL2, F2, BLOC1S6, AP3D1, GP1BB, ANO6, CD36, and ITGB3) and new suspected variants (GFI1B, FGA, GP1BA, and ITGA2B) in 11 patients. The role of NGS in patients with suspicious bleeding symptoms is growing and it changes the diagnostic algorithm. The greatest disadvantage of NGS, aside from the cost, is the occurrence of gene variants of uncertain significance.     

柔性硬质纳米复合涂层

研究了利用磁控溅射方法制备的柔性硬质纳米复合涂层。结果表明柔性硬质纳米复合涂层具有以下优异性能:是一类具有高硬度、高韧性以及抗裂纹性能的新型涂层;具有较高的硬度模量比(H/E*≥0.1,E*=E/(1-ν~2))、弹性恢复系数(We≥60%)、压应力(σ0)L,且少缺陷的微观结构;生长处于Thornton结构区域相图的T区。磁控溅射非常适合制备纳米复合涂层,文中将对其制备柔性纳米复合薄膜的机理做深入阐述。涂层生长主要受以下3个参数影响:涂层生长过程中吸收的能量Ep,其包含沉积原子携带的能量E_(ca)和轰击离子携带等能量E_(bi)(E_p=E_(ca)+E_(bi)),基体温度Ts和涂层材料的熔点T_m。柔性硬质涂层具有广泛的应用前景,如柔性保护涂层、柔性功能涂层、防脆性涂层开裂的柔性保护涂层以及柔性多层涂层。文中还将详细阐述低温磁控溅射制备柔性纳米复合涂层的原理,并阐述纳米复合涂层及其性能的发展趋势。     

Mechanical properties of wollastonite‐reinforced polypropylene composites modified with SEBS and SEBS‐g‐MA elastomers

Mechanical properties of isotactic polypropylene/wollastonite/styrene rubber block copolymers (iPP/wollastonite/SRBC) composites were studied as a function of elastomeric poly(styrene‐b‐ethylene‐co‐butylene‐b‐styrene) triblock copolymer (SEBS) and SEBS grafted with maleic anhydride (SEBS‐g‐MA) content from 0 to 20 vol%. Microphase morphology was stronger influenced by SRBC elastomers than by different wollastonite types. Higher encapsulation ability of SEBS‐g‐MA than SEBS caused more expressive core‐shell morphology and consequently higher notched impact strength as well as yield parameters, but lower Young's modulus. Higher ductility of the composites with SEBS than with SEBS‐g‐MA has been primarily caused by better miscibility of the polypropylene chains with SEBS molecules. Surface properties of components and adhesion parameters also indicated that adhesion at SEBS‐g‐MA/wollastonite interface, which was stronger than the one at the SEBS/wollastonite interface, influenced higher encapsulation of wollastonite particles by SEBS‐g‐MA. POLYM. ENG. SCI., 47:1873–1880, 2007. © 2007 Society of Plastics Engineers     

Characterization of 4,6‐Diazido‐N‐nitro‐1,3,5‐triazine‐2‐amine

4,6‐Diazido‐N‐nitro‐1,3,5‐triazine‐2‐amine (DANT) was prepared with a 35 % yield from cyanuric chloride in a three step process. DANT was characterized by IR and NMR spectroscopy (¹H, ¹³C, ¹⁵N), single‐crystal X‐ray diffraction, and DTA. The crystal density of DANT is 1.849 g cm⁻³. The cyclization of one azido group and one nitrogen atom of the triazine group giving tetrazole was observed for DANT in a dimethyl sulfoxide solution using NMR spectroscopy. An equilibrium exists between the original DANT molecule and its cyclic form at a ratio of 7 : 3. The sensitivity of DANT to impact is between that for PETN and RDX, sensitivity to friction is between that for lead azide and PETN, and sensitivity to electric discharge is about the same as for PETN. DANT′s heat of combustion is 2060 kJ mol⁻¹.     

Polypropylene/silica micro‐ and nanocomposites modified with poly(styrene‐b‐ethylene‐co‐butylene‐b‐styrene)

The effects of different silica loadings and elastomeric content on interfacial properties, morphology and mechanical properties of polypropylene/silica 96/4 composites modified with 5, 10, 15, and 20 vol % of poly(styrene‐b‐ethylene‐co‐butylene‐b‐styrene) SEBS added to total composite volume were investigated. Four silica fillers differing in size (nano‐ vs. micro‐) and in surface properties (untreated vs. treated) were chosen as fillers. Elastomer SEBS was added as impact modifier and compatibilizer at the same time. The morphology of ternary polymer composites revealed by light and scanning electron microscopies was compared with morphology predicted models based on interfacial properties. The results indicated that general morphology of composite systems was determined primarily by interfacial properties, whereas the spherulitic morphology of polypropylene matrix was a result of two competitive effects: nucleation effect of filler and solidification effect of elastomer. Tensile and impact strength properties were mainly influenced by combined competetive effects of stiff filler and tough SEBS elastomer. Spherulitic morphology of polypropylene matrix might affect some mechanical properties additionally. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 41486.     

‘Land for infrastructure’ in Ho Chi Minh City: land-based financing of transportation improvement

This paper explores how land-based financing mechanisms are currently used in Ho Chi Minh City as a public-private funding strategy. The Land-for-Infrastructure (LFI) mechanism appears as a solution to produce infrastructures. We found that the implementation of the LFI mechanism remains difficult, but eventually can lead to success. By ‘trial-error-transcend,’ the City managed to build two essential roads while the developers received attractive investment opportunities in urban development. This mechanism cannot, however, be seen as a panacea for the local authorities due to constraints to replicate it and potential undesired ‘side effects.’     

Properties of tin/plasma polymer nanocomposites

Thin composite layers (tin in plasma polymer matrix) were prepared in a stainless steel vacuum chamber. An RF powered magnetron with tin target was used to excite the discharge and to activate the monomer species (n-hexane). The gas mixture introduced comprised Ar and n-hexane vapours. The properties of the films and chemical composition were characterized by AFM (surface morphology), TEM and Electron tomography (bulk structure characterization), XPS and FTIR spectroscopy (chemical composition analyses). Current-voltage characteristics were measured to examine the electrical properties of the layers and their dependence on the deposition parameters.