Ribosomal Synthesis of Natural‐Product‐Like Bicyclic Peptides in Escherichia coli

Methods to access natural‐product‐like macrocyclic peptides can disclose new opportunities for the exploration of this important structural class for chemical biology and drug discovery applications. Here, the scope and mechanism of a novel strategy for directing the biosynthesis of thioether‐bridged bicyclic peptides in bacterial cells was investigated. This method entails split intein‐catalyzed head‐to‐tail cyclization of a ribosomally produced precursor peptide, combined with inter‐side‐chain crosslinking through a genetically encoded cysteine‐reactive amino acid. This strategy could be successfully applied to achieve formation of structurally diverse bicyclic peptides with high efficiency and selectivity in Escherichia coli. Insights into the sequence of reactions underlying the peptide bicyclization process were gained from time‐course experiments. Finally, the potential utility of this methodology toward the discovery of macrocyclic peptides with enhanced functional properties was demonstrated through the isolation of a bicyclic peptide with sub‐micromolar affinity for streptavidin.     

The kinetic evidence for the formation of multiple active species in a bis(phenoxy-imine) zirconium dichloride/MAO catalyst during ethylene polymerization

The effects of polymerization time and temperature on the molecular weight and molecular weight distribution of polyethylene, produced over homogeneous catalyst bis[N-(3-tert-butyl salicylidene)anilinato]zirconium(IV) dichloride ^tBu-L₂ZrCl₂/MAO have been studied. The data on the number of active centers (C_P) and propagation rate constants (k_P) at different polymerization time have been obtained as well. It was found that at a short polymerization time two types of active centers, producing low molecular weight PE (M_w = (4-10) × 10³ g mol⁻¹) are formed. The number of these centers was estimated to be 11% of total zirconium complex and their reactivity is very high (the k_P value was found to be 54 × 10³ L mol⁻¹ s⁻¹ at 35 °C). High initial activity of the catalyst fell with the increase in polymerization time, whereas the polydispersity values of the resulting PE increase due to formation of new centers, producing high molecular weight PE (M_w = (30-1300) × 10³ g mol⁻¹). It was found that the decrease in activity is caused by reducing the initial active centers number and lower reactivity of the new-formed centers (k_P = 17 × 10³ L mol⁻¹ s⁻¹).     

直流和脉冲工作的VDMOS可靠性试验

基于恒定电应力温度斜坡法(CETRM),对工作于直流状态和脉冲状态下的VDMOS功率器件进行可靠性研究,考察了器件阈值电压、跨导以及导通电阻的退化情况,得出在两种工作状态下均是跨导为失效敏感参数。在直流工作状态下,VDMOS失效激活能为0.57~0.68 eV,寿命为7.97×105~1.15×107h;在脉冲工作状态下,VDMOS失效激活能为0.66~0.7 eV,寿命为4.3×105~4.6×106h。对跨导的退化机理进行了分析。     

Adsorption of Positively-Charged β-Lactoglobulin Derivatives to Casein Micelles

Positively-charged β-lactoglobulin derivatives prepared by amidation or esterification of available carboxyl groups of the native protein were bound strongly by casein micelles. Increasing amounts of these additives progressively decreased rennet coagulation time of casein micelles. Higher concentrations of positively-charged proteins coagulated casein micelles without addition of rennet extract. Of the modified proteins tested, approximately 1.0 g amidated β-lactoglobulin, 2.0 g ethyl-esterified β-lactoglobulin, or 1.0 g methyl-esterified β-lactoglobulin would be required to coagulate 100 ml of casein micelles at concentrations in milk.     

Analysis of intraparticle diffusion on adsorption of crystal violet on bentonite

Abstract

In the present work, kinetics of crystal violet (CV) adsorption on bentonite was studied by pore volume and surface diffusion model (PVSDM), surface diffusion model (SDM), and pore volume diffusion model (PVDM). The adsorption decay curves were obtained in batch system using different adsorbent dosages. The PVDM model did not interpreted the kinetic adsorption since the calculated value of D_p equal to 5.64?×?10^?7 cm² s^?1 predicted a slower adsorption than that obtained by the experimental data. The PVSDM results indicates that the intraparticle diffusion is predominantly due to surface diffusion (93%) and the pore volume diffusion can be negligible. Once the surface diffusion was the limiting step, the estimation with one (D_s) and two (D_sq and α) parameters were tested in the SDM model. The statistical analysis revealed that the one-parameter SDM model was most appropriate to predict the CV adsorption on bentonite. The optimal values of Ds ranged from 6.19?×?10^?10 to 6.49?×?10^?10 cm² s^?1, and decrease with the adsorbent dosage.     

Investigating the Role of Conformational Effects on Laccase Stability and Hyperactivation under Stress Conditions

Fungal laccase from Steccherinum ochraceum 1833 displays remarkable stability under different harsh conditions: organic/buffer mixtures, thermal treatment, and microwave radiation. The behavior is particularly significant in the light of the sharp inactivation observed for two different fungal laccases. Laccase from S. ochraceum 1833 also displays hyperactivation under mild thermal treatment (60 °C). Molecular dynamics simulations at 80 °C explained how this laccase retains the geometry of the electron transfer pathway, thereby assuring electron transfer through the copper ions and thus maintaining its catalytic activity at high temperature. Spectroscopic studies revealed that the thermal activation corresponds to specific conformational changes in the protein. The results indicate that this laccase is potentially applicable under denaturing conditions that might be beneficial for the biotransformation of recalcitrant substrates.     

Cyanine Dyes for Photo-Thermal Therapy: A Comparison of Synthetic Liposomes and Natural Erythrocyte-Based Carriers

Cyanine fluorescent dyes are attractive diagnostic or therapeutic agents due to their excellent optical properties. However, in free form, their use in biological applications is limited due to the short circulation time, instability, and toxicity. Therefore, their encapsulation into nano-carriers might help overcome the above-mentioned issues. In addition to indocyanine green (ICG), which is clinically approved and therefore the most widely used fluorescent dye, we tested the structurally similar and cheaper alternative called IR-820. Both dyes were encapsulated into liposomes. However, due to the synthetic origin of liposomes, they can induce an immunogenic response. To address this challenge, we proposed to use erythrocyte membrane vesicles (EMVs) as “new era” nano-carriers for cyanine dyes. The optical properties of both dyes were investigated in different biological relevant media. Then, the temperature stability and photo-stability of dyes in free form and encapsulated into liposomes and EMVs were evaluated. Nano-carriers efficiently protected dyes from thermal degradation, as well as from photo-induced degradation. Finally, a hemotoxicity study revealed that EMVs seem less hemotoxic dye carriers than clinically approved liposomes. Herein, we showed that EMVs exhibit great potential as nano-carriers for dyes with improved stability and hemocompatibility without losing excellent optical properties.     

The ritual as a psychotherapeutic intervention.

Examines ritual as a tool for psychotherapeutic application with clients experiencing grief, trauma, anxiety, and family dysfunction. Case vignettes are provided to illustrate the therapeutic properties and creative power of rituals as interventions. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Media,Technologies, Cooperation – Rethinking Publics and Publicness in the MENA Region

Computer Supported Cooperative Work (CSCW) -     

Characterization of Competitive Inhibitors of Plasmodium falciparum cGMP-Dependent Protein Kinase**

Plasmodium falciparum cGMP-dependent protein kinase (PfPKG) is an enticing antimalarial drug target. Novel chemotypes are needed because existing inhibitors have safety issues that may prevent further development. This work demonstrates isoxazole-based compounds are potent ATP competitive inhibitors of PfPKG and discloses a new analogue in this series. Isoxazoles 3 and 5 had K_i values that are comparable to a known standard, 4-[2-(4-fluorophenyl)-5-(1-methylpiperidine-4-yl)-1H pyrrol-3-yl] pyridine. They also exhibited excellent selectivity for PfPKG over the human orthologue and the gatekeeper mutant T618Q PfPKG, which mimics the less accessible binding site of the human orthologue. The human orthologue's larger binding site volume is predicted to explain the selectivity of the inhibitors for the P. falciparum enzyme.