A procedure to prepare cross-sectional samples for TEM

Cross-sectional transmission electron microscopy (XTEM) is an imaging technique particularly suited to the study of layered structures. For integrated electronic devices it has become a common practice to use XTEM to assess the shape and crystallinity of component layers as well as defect structures introduced by processing. A procedure for preparing samples to be viewed by XTEM is described.     

A novel combined electrochemical-magnetic method for water treatment

Electrocoagulation (EC) is a wastewater treatment process in which aqueous pollutants can be removed by adsorption, entrapment, precipitation or coalescence during a coagulation step produced by electrochemically generated metallic species. When using Fe as the sacrificial electrode, Fe(2+) and Fe(3+) ions are formed. As Fe(3+) species are paramagnetic, this property can in principle be used to facilitate their removal through the application of a magnetic field. In the present work we present a proof-of-concept for a combined electrochemical-magnetic method for pollutant removal. For this approach, the amounts of Fe(2+) and Fe(3+) produced in an EC cell at various voltages were measured by spectroscopic methods to confirm that Fe(3+) species predominate (up to 84%). The effectiveness of the presence of a magnetic field in the precipitation of coagulants from a suspension was confirmed by monitoring the turbidity change versus time with and without exposure to a magnetic field, up to a 30% improvement.     

Sleeping Beauties in Psychology: Comparisons of "Hits" and "Missed Signals" in Psychological Journals.

Scientific publications tend to be forgotten quickly. A few works, however, are still cited 100 years and more after their publication. The author used bibliometric methods to compare "hits" (works noticed by the scientific community soon after their publication) with "missed signals" (works that went unnoticed until much later) by investigating 2 psychological journals founded in the 1890s: Zeitschrift für Psychologie and Psychological Review. All articles that were published in either of these journals up to 1920 and cited more than 25 times in the Web of Science up to the year 2000 were considered for inclusion in the analysis. It emerged that hits corresponded more closely to the focus of scientific attention at the time of publication than missed signals. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The Effect of the Length of To-Be-Remembered Lists and Intervening Lists on Free Recall: A Reexamination Using Overt Rehearsal.

In 3 experiments, the authors investigated the effects of to-be-remembered (TBR) and intervening list length on free recall to determine whether selective rehearsal could explain the previous finding that recall was affected only by TBR list length. In Experiments 1 (covert rehearsal) and 2 (overt rehearsal), participants saw 5- and 20-word lists and had to recall the list prior to that last presented list. In Experiment 3, either 1 or 2 lists were presented, and recall of TBR list was postcued. Recall proportion decreased with increased TBR list length. Moreover, the authors found extended recency effects when recall was replotted by when words were last rehearsed (Experiments 2 and 3) and an effect of intervening list length when rehearsal was reduced (Experiment 3). (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Influence of Different Salts on the G-Quadruplex Structure Formed from the Reversed Human Telomeric DNA Sequence

G-rich telomeric DNA plays a major role in the stabilization of chromosomes and can fold into a plethora of different G-quadruplex structures in the presence of mono- and divalent cations. The reversed human telomeric DNA sequence (5′-(GGG ATT)₄; RevHumTel) was previously shown to have interesting properties that can be exploited for chemical sensing and as a chemical switch in DNA nanotechnology. Here, we analyze the specific G-quadruplex structures formed by RevHumTel in the presence of K⁺, Na⁺, Mg²⁺ and Ca²⁺ cations using circular dichroism spectroscopy (CDS) and Förster resonance energy transfer (FRET) based on fluorescence lifetimes. CDS is able to reveal strand and loop orientations, whereas FRET gives information about the distances between the 5′-end and the 3′-end, and also, the number of G-quadruplex species formed. Based on this combined information we derived specific G-quadruplex structures formed from RevHumTel, i.e., a chair-type and a hybrid-type G-quadruplex structure formed in presence of K⁺, whereas Na⁺ induces the formation of up to three different G-quadruplexes (a basket-type, a propeller-type and a hybrid-type structure). In the presence of Mg²⁺ and Ca²⁺ two different parallel G-quadruplexes are formed (one of which is a propeller-type structure). This study will support the fundamental understanding of the G-quadruplex formation in different environments and a rational design of G-quadruplex-based applications in sensing and nanotechnology.     

Exploiting ELIOT for Scaffold-Repurposing Opportunities: TRIM33 a Possible Novel E3 Ligase to Expand the Toolbox for PROTAC Design

The field of targeted protein degradation, through the control of the ubiquitin–proteasome system (UPS), is progressing considerably; to exploit this new therapeutic modality, the proteolysis targeting chimera (PROTAC) technology was born. The opportunity to use PROTACs engaging of new E3 ligases that can hijack and control the UPS system could greatly extend the applicability of degrading molecules. To this end, here we show a potential application of the ELIOT (E3 LIgase pocketOme navigaTor) platform, previously published by this group, for a scaffold-repurposing strategy to identify new ligands for a novel E3 ligase, such as TRIM33. Starting from ELIOT, a case study of the cross-relationship using GRID Molecular Interaction Field (MIF) similarities between TRIM24 and TRIM33 binding sites was selected. Based on the assumption that similar pockets could bind similar ligands and considering that TRIM24 has 12 known co-crystalised ligands, we applied a scaffold-repurposing strategy for the identification of TRIM33 ligands exploiting the scaffold of TRIM24 ligands. We performed a deeper computational analysis to identify pocket similarities and differences, followed by docking and water analysis; selected ligands were synthesised and subsequently tested against TRIM33 via HTRF binding assay, and we obtained the first-ever X-ray crystallographic complexes of TRIM33α with three of the selected compounds.     

Extraction of pure lycopene from industrial tomato by‐products in water using a new high‐pressure process

BACKGROUND: Lycopene, a precursor of β‐carotene with a well‐known antioxidant activity, contained in many natural products such as tomato (Lycopersicon esculentum Mill.), watermelon, red pepper and papaya, is usually recovered from natural vegetal sources using organic solvents and a purification step. In this paper an innovative process for the extraction of pure lycopene from tomato waste in water that uses the Naviglio^® extractor and water as extracting phase is presented. RESULTS: Lycopene was obtained in the all‐trans form at a very high grade of purity, not less than 98% (w/w), with an average recovery of 14% (w/w). The availability of high‐purity trans‐lycopene allowed measurement of the molar absorption coefficient. An alternative procedure for high‐performance liquid chromatographic analysis using a phenyl‐hexyl silicone phase as inverse phase and a linear gradient in water and acetonitrile is also described. CONCLUSIONS: The use of water as extracting phase considerably reduces the cost of the entire process when compared with the commonly used solvent‐based procedure or with the newer supercritical extraction process of lycopene from tomato waste. Lycopene, not soluble in water, was recovered in a quasi‐crystalline solid form and purified by solid‐phase extraction using a small amount of organic solvent. Copyright © 2008 Society of Chemical Industry This article was published online on September 15, 2008. Errors in Figures 2 ‐ 4 were subsequently identified. The publishers wish to apologise for these errors. This notice is included in the online and print versions to indicate that both have been corrected [September 19, 2008]     

Quaternary Structure and Hetero-Oligomerization of Recombinant Human Small Heat Shock Protein HspB7 (cvHsp)

In this study, a reliable and simple method of untagged recombinant human HspB7 preparation was developed. Recombinant HspB7 is presented in two oligomeric forms with an apparent molecular weight of 36 kDa (probably dimers) and oligomers with an apparent molecular weight of more than 600 kDa. By using hydrophobic and size-exclusion chromatography, we succeeded in preparation of HspB7 dimers. Mild oxidation promoted the formation of large oligomers, whereas the modification of Cys 126 by iodoacetamide prevented it. The deletion of the first 13 residues or deletion of the polySer motif (residues 17–29) also prevented the formation of large oligomers of HspB7. Cys-mutants of HspB6 and HspB8 containing a single-Cys residue in the central part of the β7 strand in a position homologous to that of Cys137 in HspB1 can be crosslinked to the wild-type HspB7 through a disulfide bond. Immobilized on monoclonal antibodies, the wild-type HspB6 interacted with the wild-type HspB7. We suppose that formation of heterodimers of HspB7 with HspB6 and HspB8 may be important for the functional activity of these small heat shock proteins.     

Platelet-Derived PCSK9 Is Associated with LDL Metabolism and Modulates Atherothrombotic Mechanisms in Coronary Artery Disease

Platelets play a significant role in atherothrombosis. Proprotein convertase subtilisin/kexin type 9 (PCSK9) is critically involved in the regulation of LDL metabolism and interacts with platelet function. The effect of PCSK9 in platelet function is poorly understood. The authors of this article sought to characterize platelets as a major source of PCSK9 and PCSK9’s role in atherothrombosis. In a large cohort of patients with coronary artery disease (CAD), platelet count, platelet reactivity, and platelet-derived PCSK9 release were analyzed. The role of platelet PCSK9 on platelet and monocyte function was investigated in vitro. Platelet count and hyper-reactivity correlated with plasma LDL in CAD. The circulating platelets express on their surface and release substantial amounts of PCSK9. Release of PCSK9 augmented platelet-dependent thrombosis, monocyte migration, and differentiation into macrophages/foam cells. Platelets and PCSK9 accumulated in tissue derived from atherosclerotic carotid arteries in areas of macrophages. PCSK9 inhibition reduced platelet activation and platelet-dependent thrombo-inflammation. The authors identified platelets as a source of PCSK9 in CAD, which may have an impact on LDL metabolism. Furthermore, platelet-derived PCSK9 contributes to atherothrombosis, and inhibition of PCSK9 attenuates thrombo-inflammation, which may contribute to the reported beneficial clinical effects.